TW201311907A - Genomic diagnostics using circulating endothelial cells - Google Patents

Genomic diagnostics using circulating endothelial cells Download PDF

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TW201311907A
TW201311907A TW101124348A TW101124348A TW201311907A TW 201311907 A TW201311907 A TW 201311907A TW 101124348 A TW101124348 A TW 101124348A TW 101124348 A TW101124348 A TW 101124348A TW 201311907 A TW201311907 A TW 201311907A
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Taiwan
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human
ena
ribonucleic acid
message
cec
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TW101124348A
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Yixin Wang
Bao-Ying Huang
Jack Yu
Yuqui Jiang
Tim Jatkoe
Yashoda Rajpurohit
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Veridex Llc
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

Abstract

Circulating Endothelial Cells are isolated from patient blood and gene expression of the cells is analyzed to assess a medical condition or the tissue of origin of the cell. Kits for conducting the method are also provided.

Description

使用循環內皮細胞的基因診斷 Genetic diagnosis using circulating endothelial cells

循環內皮細胞係從病患血液中單離，然後分析該細胞的基因表現，以便評估醫療狀況或該細胞的來源組織。亦提供執行該方法適用的套件。 The circulating endothelial cell line is isolated from the blood of the patient and the gene expression of the cell is then analyzed to assess the medical condition or the tissue of the cell. A kit for performing this method is also available.

循環內皮細胞(CEC)在健康人體內的出現數量低，但在多種人類疾病，包括心血管疾病和癌症患者體內已經觀察到其數量增加現象。CEC的特徵將有助於瞭解和監測這些疾病和其他狀況。CEC係可採用各種技術，包括以CD 146抗體和磁分離進行的抗體擷取以及流式細胞儀和其他方式，而將其從外周血液中單離出來。不幸的是，CEC分離和分析因受到白血球大量存在而變得複雜。對CEC識別有幫助的是讓其與重要因子，例如其原始組織有關聯，提供一個基礎進一步對其進行分析，並根據該分析提供相關的醫療訊息。 Circulating endothelial cells (CEC) are present in low numbers in healthy individuals, but their number has been observed in a variety of human diseases, including cardiovascular diseases and cancer patients. The characteristics of the CEC will help to understand and monitor these diseases and other conditions. CEC can be isolated from peripheral blood by a variety of techniques, including antibody capture with CD 146 antibodies and magnetic separation, and flow cytometry and other means. Unfortunately, CEC separation and analysis is complicated by the large presence of white blood cells. Helping CEC identification is to associate it with important factors, such as its original organization, to provide a basis for further analysis and to provide relevant medical information based on the analysis.

在本發明一態樣中，CEC係單離的，然後分析CEC基因表現以便評估醫療病症。較佳地，基因表現係採用微陣列分析或例如反轉錄PCR(RTPCR)的放大和識別方法進行。 In one aspect of the invention, the CEC is isolated and then analyzed for CEC gene expression to assess a medical condition. Preferably, the gene expression is performed using microarray analysis or amplification and recognition methods such as reverse transcription PCR (RTPCR).

在本發明另一態樣中，從130個特異基因之族群中選出在外周血液單核細胞(PBMC)中表現低，而在血 管內皮細胞中表現高的基因，然後將該基因裝配成CEC標記。 In another aspect of the invention, a population of 130 specific genes is selected to be low in peripheral blood mononuclear cells (PBMC), but in blood A gene that expresses high in endothelial cells and then assembles the gene into a CEC marker.

在本發明又另一態樣中，基因為基礎的CEC標記為與以下一個或多個相關者：細胞運動、細胞遷移、血管生成或細胞附著。較佳是這類標記相對於其他細胞為過度表現。 In yet another aspect of the invention, the gene-based CEC marker is associated with one or more of the following: cell motility, cell migration, angiogenesis, or cell attachment. Preferably such markers are overexpressed relative to other cells.

在本發明又一進一步態樣中，基因為基礎的標記視不同血管類型有不同表現，而能夠識別所擷取CEC的原始血管或組織。較佳為，該標記係從67個基因中選出在CEC上相對於其他細胞為過度表現者。 In yet a further aspect of the invention, the gene-based marker has different manifestations depending on the type of blood vessel and is capable of identifying the original blood vessel or tissue from which the CEC is drawn. Preferably, the marker is selected from 67 genes that are overexpressed on CEC relative to other cells.

在本發明又一進一步態樣中，細胞擷取係用來獲得基因表現分析用的CEC。較佳為，利用免疫磁性法進行擷取，然後透過分析提供醫療評估，例如疾病或狀況診斷、預測或預後。 In yet another aspect of the invention, the cell retrieval system is used to obtain a CEC for gene expression analysis. Preferably, the immunomagnetic method is used for the extraction and then the analysis provides a medical assessment, such as a disease or condition diagnosis, prediction or prognosis.

在本發明又一進一步態樣中，提供有CEC分析套件。較佳為，該套件包括CEC基因表現識別和分析用的試劑。此外，該套件可含有單離CEC用的擷取試劑。該套件又可包括機械碼實施例，其中該機械碼實施例將訊息和演算法應用至CEC單離和/或分析實施期間所產生的訊息上，因而產生或促進醫療評估。 In yet another aspect of the invention, a CEC analysis kit is provided. Preferably, the kit includes reagents for CEC gene expression recognition and analysis. In addition, the kit can contain a draw reagent for separation from CEC. The kit, in turn, can include a mechanical code embodiment, wherein the mechanical code embodiment applies the message and algorithm to the message generated during the CEC separation and/or analysis implementation, thereby generating or facilitating a medical assessment.

本發明提供將內皮細胞快速有效率地從生物樣品中單離和鑑定的組成、方法和套件。該方法描述血液 樣品中CEC的單離和鑑定，同時將非特異性鍵結或截留的細胞選擇性降至最低。 The present invention provides compositions, methods and kits for the rapid and efficient isolation and identification of endothelial cells from biological samples. This method describes blood The isolation and identification of CEC in the sample while minimizing the selectivity of non-specifically bound or trapped cells.

雖然單離、濃縮和分析血液中CEC適用的任何有效機制，係可用來擷取和濃縮分析用的CEC，但是其收集的較佳方法是結合免疫磁性法濃縮技術和免疫螢光標示技術，再搭配適當的分析平台而成。相關測試具有靈敏度和特異性，以檢測全血樣品中的該等稀有細胞，並利用這些稀有細胞進行疾病和狀況的臨床過程分析，以及包括預測和預後之許多相同態樣的相關評估。 Although any effective mechanism for the isolation, concentration, and analysis of CEC in blood can be used to extract and concentrate CEC for analysis, the preferred method of collection is to combine immunomagnetic condensation techniques with immunofluorescence labeling techniques. Made with the appropriate analysis platform. Relevant tests have sensitivity and specificity to detect such rare cells in whole blood samples, and to use these rare cells for clinical process analysis of diseases and conditions, as well as related assessments of many identical aspects including prediction and prognosis.

較佳實施例中所使用的擷取和分離技術係已廣泛用於分析循環腫瘤細胞(CTC)，藉由使用例如一種工具探究癌症患者外周循環中的上皮原始細胞的顯著性而完成。這項技術係如美國專利第6,365,362號和美國專利第6,645,731中所述。使用這種技術的「CellSearch」系統(新澤西州力登的Veridex公司)是一種根據從血液單離之細胞的螢光顯微鏡術之自動系統。這在擷取和單離CEC也是已知的用途。該系統包含一整合的電腦控制螢光顯微鏡和配有磁軛套件的自動化平台，其中該磁軛套件將持有一拋棄式樣品盒。該磁軛係設計成能讓磁性液體所標示的候選稀有細胞位於樣品室內，以對進行樣品盒上方觀測表面進行局部磁化，便於顯微鏡下觀看。軟體會檢測出抗體所標示且含有來自血液中之內皮細胞的細胞。在一較佳實施例中，例 如「CellSave」細胞防腐劑的一種防腐劑，係用在7.5毫升全血中來單離出有興趣的細胞。添加細胞特異性磁性粒子後，較佳為培養約20分鐘。CellSave防腐劑係可設置在例如血液樣品收集的管內，或可作為本發明套件其中一部分。磁分離後，鍵結到免疫磁性連結抗體的細胞係受到磁性而貼附於管壁。接著抽吸出未鍵結的樣品，然後添加等張溶液來再懸浮樣品。讓一種核酸染料、對應於指定標記的單株抗體和CD 45(一種寬光譜白血球標記)連同樣品一起培養。磁分離後，再度抽吸出未鍵結的部份，讓鍵結和標示細胞再懸浮在0.2毫升等張溶液中。讓該樣品懸浮在一細胞展示室中並放置於磁場設備內，其中該磁場設備內的磁場會讓該磁性標示細胞定向成符合CellSearch系統中螢光顯微檢查。採用系統所列舉的對照細胞，和呈現給操作者進行清單列舉以識別這類態樣之形態的候選目標細胞，即可自動識別出細胞。接著即可讓擷取的細胞進行本發明基因為基礎的分析。 The extraction and separation techniques used in the preferred embodiments have been widely used to analyze circulating tumor cells (CTCs) by using, for example, a tool to explore the significance of epithelial blast cells in the peripheral circulation of cancer patients. This technique is described in U.S. Patent No. 6,365,362 and U.S. Patent No. 6,645,731. The "CellSearch" system (Veridex, Raritan, NJ) using this technology is an automated system based on fluorescence microscopy of cells isolated from blood. This is also a known use in capturing and separating CEC. The system includes an integrated computer-controlled fluorescent microscope and an automated platform with a yoke kit that will hold a disposable sample cartridge. The yoke is designed such that the candidate rare cells labeled by the magnetic liquid are located in the sample chamber to locally magnetize the observation surface above the sample cartridge for viewing under the microscope. The software detects cells that are labeled by antibodies and contain endothelial cells from the blood. In a preferred embodiment, an example A preservative such as "CellSave" cell preservative is used in 7.5 ml of whole blood to separate cells of interest. After the addition of the cell-specific magnetic particles, it is preferably cultured for about 20 minutes. The CellSave preservative can be placed, for example, in a tube for blood sample collection, or can be part of a kit of the invention. After magnetic separation, the cell line bonded to the immunomagnetic-linked antibody is magnetically attached to the tube wall. The unbound sample is then aspirated and an isotonic solution is added to resuspend the sample. A nucleic acid dye, a monoclonal antibody corresponding to the indicated label, and CD 45 (a broad spectrum white blood cell marker) are incubated with the sample. After magnetic separation, the unbound portion was again aspirated, and the bonded and labeled cells were resuspended in 0.2 ml isotonic solution. The sample is suspended in a cell display chamber and placed in a magnetic field device wherein the magnetic field within the magnetic field device directs the magnetic indicator cell to conform to the fluorescence microscopy in the CellSearch system. The cells are automatically identified using control cells enumerated by the system, and candidate target cells presented to the operator for enumeration to identify the morphology of such a pattern. The captured cells can then be subjected to the gene-based assay of the present invention.

進行CEC單離所適用試劑中所包括的較佳磁性粒子為表現成膠體的粒子。這種粒子特點為次微米的粒子大小、這通常為小於約200奈米(0.20微米)，和從溶液中重力分離出後維持長時間的穩定性。除許多其他優勢以外，還有，此大小範圍使其在細胞分析所運用的分析技術中幾乎是觀察不到。在90-150 nm範圍內且具有介於70-90%磁質量的粒子，亦被預期可用於 本發明。適合的磁性粒子是由被分子圍著的超順磁性材料之一結晶核心組成，該等分子係被結合，例如被物理吸附或被共價連接至磁心且給予安定膠態性質。塗層材料較佳係應施作達一個數量，才得以有效避免樣品中所發現的生物大分子和磁性核心間發生非特異性相互作用。這種生物大分子可包括例如非目標細胞表面上之唾液酸殘留的碳水化合物、凝集素、醣蛋白和其他膜成分。此外，該材料每奈米粒子應盡可能包含越高的磁性質量。包含核心之磁性結晶的大小為足夠小到不含有一完整的磁域。奈米粒子的尺寸小到足以使其布朗能量超過其磁矩。因此，北極、南極對準以及接著發生之這些膠態磁性粒子的相互吸引/排斥，看來並不像是有發生，即使是在影響其等之溶液安定性之適度的強磁埸中。最後，磁性粒子應該可以在高磁力梯度外場分離器中分離。該特性有助於試樣處理且提供比裝滿鐵磁珠或鋼絲絨的更複雜內梯度管柱更好的經濟優勢。具有上述性質的磁性粒子係可藉由美國專利第4,795,698、5,597,531和5,698,271號中所述之基底材料修正例製成。 Preferred magnetic particles included in the reagent for performing CEC separation are particles which exhibit a colloid. Such particles are characterized by submicron particle sizes, which are typically less than about 200 nanometers (0.20 micrometers), and maintain long-term stability after gravity separation from solution. In addition to many other advantages, this size range makes it almost impossible to observe in the analytical techniques used in cell analysis. Particles in the range of 90-150 nm with a magnetic mass between 70-90% are also expected to be used this invention. Suitable magnetic particles consist of a crystalline core of one of the superparamagnetic materials surrounded by molecules that are bound, for example physically adsorbed or covalently attached to the core and imparting a stable colloidal nature. Preferably, the coating material is applied in an amount to effectively prevent non-specific interactions between the biomacromolecule and the magnetic core found in the sample. Such biomacromolecules may include, for example, carbohydrates, lectins, glycoproteins, and other membrane components that are residual of sialic acid on the surface of non-target cells. In addition, the material should contain as high a magnetic mass as possible per nanometer particle. The size of the magnetic crystal containing the core is small enough to not contain a complete magnetic domain. The size of the nanoparticle is small enough that its Brownian energy exceeds its magnetic moment. Therefore, the mutual attraction/rejection of the colloidal magnetic particles aligned with the north pole, the south pole, and the subsequent occurrence of the colloidal magnetic particles does not appear to occur, even in a moderately strong magnetic field that affects the stability of the solution. Finally, the magnetic particles should be separable in a high magnetic gradient external field separator. This feature facilitates sample processing and provides a better economic advantage than a more complex internal gradient column filled with ferromagnetic beads or steel wool. The magnetic particles having the above properties can be produced by the substrate material modification described in U.S. Patent Nos. 4,795,698, 5,597,531 and 5,698,271.

免疫磁性樣品的製備，對於減少樣品量和獲得104倍濃縮後目標細胞而言非常重要。本發明較佳套件中所使用的試劑包括：對應於泛白血球抗原CD 45以識別白血球(非目標細胞)的一抗體；能夠排除殘餘紅血球細胞、血小板和其他無核物件的一細胞類型特異 或核酸染料；用在對應於目標細胞結構的一生物特異試劑或抗體，或針對目標膜具有特異性，與用來免疫磁性選擇細胞不同的一抗體。 Immunomagnetic sample preparation is very important to reduce the amount of sample and the terms obtained after 104-fold concentration of target cells. The reagents used in the preferred kit of the present invention include: an antibody corresponding to the white blood cell antigen CD 45 to recognize white blood cells (non-target cells); a cell type specific or nucleic acid capable of excluding residual red blood cells, platelets and other non-nuclear articles Dye; used as a biospecific reagent or antibody corresponding to the structure of the target cell, or specific for the target membrane, unlike an antibody used to immunomagnetically select cells.

形態分析係可在各種分析平台上進行，包括例如CELLSPOTTER系統，它是一種採用人工細胞觀測之顯微鏡檢測器的一磁性細胞固定化和分析系統，分別如美國專利第5,876,593、5,985,153和6,136,182號中所述。上述美國專利申請案整體以引用方式併入本文，作為揭露手動或自動定量和定性細胞分析適用的各自儀器和方法。其他分析平台包括但不限於雷射掃描儀(Compucyte)、亮場基成像分析(Chromavision)和毛細管容積法(Biometric Imaging)。 Morphological analysis can be performed on a variety of analytical platforms, including, for example, the CELLSPOTTER system, which is a magnetic cell immobilization and analysis system using a microscopic detector for artificial cell observation, as in U.S. Patent Nos. 5,876,593, 5,985,153 and 6,136,182, respectively. Said. The above-identified U.S. Patent Application is hereby incorporated by reference herein in its entirety in its entirety in its entirety in the the the the the the the the Other analytical platforms include, but are not limited to, laser scanners, Chromavision, and Biometric Imaging.

本發明套件較佳包括或係可結合具有試劑的套件使用，以進行所獲得之細胞(CEC)的分子分析。這些包括有助於確定相關細胞之基因表現模式之方法的試劑，以及有助於確定基因表現之蛋白基底之方法的試劑，其中蛋白基底之方法包括有反轉錄PCR(RT-PCR)、競爭性RT-PCR、即時RT-PCR、差異顯示RT-PCR、北方墨點分析和其他相關測試。雖然可能採用個別PCR反應進行這些技術，但是最好是放大從訊息核糖核甘酸(mRNA)所產生的互補去氧核醣核酸(cDNA)或互補核糖核甘酸(cRNA)，然後透過微陣列分析。其生產適用的一些不同陣列配置和方法為熟悉該項技術領域者所知，且如下美國專利所述：5,445,934； 5,532,128；5,556,752；5,242,974；5,384,261；5,405,783；5,412,087；5,424,186；5,429,807；5,436,327；5,472,672；5,527,681；5,529,756；5,545,531；5,554,501；5,561,071；5,571,639；5,593,839；5,599,695；5,624,711；5,658,734；和5,700,637；其揭露內容以引用方式併入本文。 The kit of the invention preferably comprises or can be used in conjunction with a kit having reagents for performing molecular analysis of the obtained cells (CEC). These include reagents that aid in the determination of the gene expression pattern of the relevant cells, as well as reagents that aid in the determination of the protein substrate of the gene expression, wherein the protein substrate method includes reverse transcription PCR (RT-PCR), competition. RT-PCR, RT-PCR, differential display RT-PCR, Northern blot analysis and other related tests. While it is possible to perform these techniques using individual PCR reactions, it is preferred to amplify the complementary deoxyribonucleic acid (cDNA) or complementary ribonucleotide (cRNA) produced from the message ribonucleotide (mRNA) and then pass through a microarray analysis. Some different array configurations and methods for which production is applicable are known to those skilled in the art and are described in the following U.S. Patent: 5,445,934; 5,532,128; 5,556,752; 5,242,974; 5,384,261; 5,405,783; 5,412,087; 5,424,186; 5,429,807; 5,436,327; 5,472,672; 5,527,681; 5,529,756; 5,545,531; 5,554,501; 5,561,071; 5,571,639; 5,593,839; 5,599,695; 5,624,711; 5,658,734; and 5,700,637; Into this article.

微陣列技術能夠同時測量數千個基因的穩定狀態mRNA水平，藉此展現出一種強大工具來識別如細胞增殖失控之開始、遏止或調節的效應。有兩種微陣列技術為目前廣泛使用。第一種是cDNA陣列，第二種則是寡核苷酸陣列。雖然在這些晶片結構中有差異存在，不過基本上所有下游的數據分析和輸出係相同。這些分析的產出通常是用來檢測來自樣品之cDNA序列的標示探針所接收到之信號強度的測量值，其中該樣品會在微陣列上一個已知位置處的核酸序列混合。通常，該信號強度正比於cDNA數量和因而在樣品細胞所表現的mRNA。有大量這種技術可用且有幫助。確定基因表現適用的較佳方法係可發現於美國專利案Linsley等人的第6,271,002號；Friend等人的第6,218,122號；Peck等人的第6,218,114號；和Wang等人的第6,004,755號，各個揭露以引用方式併入本文。進行這些程序適用的成分和試劑係可包括在本發明的套件內。 Microarray technology is capable of simultaneously measuring the steady state mRNA levels of thousands of genes, thereby demonstrating a powerful tool to identify effects such as the onset, arrest or regulation of uncontrolled cell proliferation. There are two types of microarray technology that are currently widely used. The first is a cDNA array and the second is an oligonucleotide array. Although there are differences in these wafer structures, substantially all downstream data analysis and output systems are identical. The output of these assays is typically a measure of the intensity of the signal received by the labeled probe from the cDNA sequence of the sample, wherein the sample will be mixed at a known position on the microarray. Typically, the signal intensity is proportional to the amount of cDNA and thus the mRNA expressed in the sample cells. There are a large number of such technologies available and helpful. A preferred method for determining the suitability of a gene is found in U.S. Patent No. 6,271,002 to Linsley et al.; 6, 218,122 to Friend et al; 6, 218,114 to Peck et al; and 6,004,755 to Wang et al. Incorporated herein by reference. The ingredients and reagents to which these procedures are applicable may be included in the kit of the present invention.

在本發明最佳實施例中，收集病患的血液，然後使用免疫磁性擷取技術、例如在CellSearch系統中來單離CEC。接著讓CEC接受單獨分析，例如搭配RT-PCR或DNA微陣列使用。基因表現模式係依照模式識別確定和處理，其為要提供給臨床醫生和/或病人的指示性疾病診斷、預測或預後。各種基因或其表現受檢測的基因為基礎之標記的表現，與例如有沒有疾病、疾病過程或發展、染上疾病或病症的可能性以及其他預測和預後判斷之身體狀況，兩者間係可得出相關性。模式識別軟體係可用來執行這些相關性，並表明相關性是否存在。電腦指令包括用來比對檢定結果和相關表現模式，和用來提供醫療評估或在提供醫療評估時有用訊息的執行碼，該電腦指令係可以DVD形式、隨身碟或任何其他方便形式而納入本發明該套件內。 In a preferred embodiment of the invention, the patient's blood is collected and then isolated from the CEC using immunomagnetic capture techniques, such as in the CellSearch system. The CEC is then subjected to a separate analysis, such as with RT-PCR or DNA microarrays. The gene expression pattern is determined and processed in accordance with pattern recognition, which is an indicative disease diagnosis, prediction or prognosis to be provided to the clinician and/or patient. The expression of a variety of genes or their expression-based gene-based markers, and for example, the presence or absence of a disease, disease process or development, the likelihood of contracting a disease or condition, and other physical conditions of prediction and prognosis, Get relevance. A pattern recognition soft system can be used to perform these correlations and indicate if a correlation exists. Computer instructions include an execution code used to compare the results of the verification and related performance patterns, and to provide a medical assessment or useful information in the provision of a medical assessment, which may be incorporated into the form of a DVD, a flash drive or any other convenient form. Invented in the kit.

實例 Instance

在以下實例中，針對來自9個解剖位置之18個內皮細胞樣品的基因表現譜進行分析。針對在內皮細胞中呈現高表現，但在PBMC中則否的一組130個基因為基礎的標記進行識別。亦針對從CellSearch系統所濃縮之內皮細胞的這些標記進行檢測。基因為基礎的標記係很容易藉由內皮細胞所崁釘之血液樣品中的QRT-PCR進行檢測。此外，針對取自不同血管類型之 內皮細胞中表現不同的一組67個標記進行識別，其中這些標記係可用來檢測欲分析的CEC來源。 In the following examples, gene expression profiles from 18 endothelial cell samples from 9 anatomical locations were analyzed. A set of 130 gene-based markers that exhibited high expression in endothelial cells but no in PBMC were identified. These markers were also tested for endothelial cells concentrated from the CellSearch system. Gene-based marker systems are readily detected by QRT-PCR in blood samples spiked by endothelial cells. In addition, for different blood vessel types A panel of 67 markers that differ in expression in endothelial cells is identified, wherein these markers can be used to detect the source of CEC to be analyzed.

實例1：CECs Example 1: CECs

凍存的HAEC(人類主動脈內皮細胞)、HUVEC(人類臍靜脈內皮細胞)、HPAEC(人類肺動脈內皮細胞)、HMVEC-ad(人類微血管內皮細胞，成人真皮層)和HASMC(人類動脈平滑肌細胞)係從Invitrogen(加州卡爾斯巴德的Invitrogen公司)取得，而凍存的HCAEC(人類冠狀動脈內皮細胞)、HUAEC(人類臍動脈內皮細胞)、HIAEC(人類髂動脈內皮細胞)、HMVEC-C(人類心臟微血管內皮細胞)細胞則係從Lonza(德國科隆的Lonza公司)取得。凍存的HSaVEC(人類隱靜脈內皮細胞)係從PromoCell(德國海德堡的PromoCell公司)取得。內皮細胞係在37℃培養箱、濕度95%和5%二氧化碳條件下，以EGM®-2內皮細胞生長培養基-2(Lonza)進行培養。平滑肌細胞係以培養基231、補充平滑肌生長補充物(SMGS)(Invitrogen公司)進行培養。HAEC、HUVEC、HPAEC、HMVEC-ad、HCAEC、HUAEC、HMVEC-C、HSaVEC的細胞團塊係從PromoCell購得。人類血液外周白血球(PBMC)總RNA係從Clontech(加州山景城的Clontech公司)購得。 Frozen HAEC (human aortic endothelial cells), HUVEC (human umbilical vein endothelial cells), HPAEC (human pulmonary artery endothelial cells), HMVEC-ad (human microvascular endothelial cells, adult dermis) and HASMC (human arterial smooth muscle cells) Obtained from Invitrogen (Invitrogen, Carlsbad, Calif.), frozen HCAEC (human coronary endothelial cells), HUAEC (human umbilical artery endothelial cells), HIAEC (human brachial artery endothelial cells), HMVEC-C ( Human cardiac microvascular endothelial cells were obtained from Lonza (Lonza, Cologne, Germany). Frozen HSaVEC (human saphenous vein endothelial cells) was obtained from PromoCell (PromoCell, Heidelberg, Germany). The endothelial cell line was cultured in EGM®-2 endothelial cell growth medium-2 (Lonza) under a 37 ° C incubator, 95% humidity and 5% carbon dioxide. The smooth muscle cell line was cultured with medium 231, supplemented with smooth muscle growth supplement (SMGS) (Invitrogen). Cell clumps of HAEC, HUVEC, HPAEC, HMVEC-ad, HCAEC, HUAEC, HMVEC-C, HSaVEC were purchased from PromoCell. Human blood peripheral white blood cell (PBMC) total RNA was purchased from Clontech (Clontech, Inc., Mountain View, CA).

來自健康捐血者的血液係抽吸到含有EDTA的真空採血管內，然後其中4毫升以CellSearch系統(新澤西州力登的Veridex公司)處理，以便單離出CEC。此係使用CEC譜套件(Veridex)，根據製造商指南使用。在崁釘實驗中，500或1000個內皮細胞(HAEC、HPAEC、HUVEC或HMVEC-ad)係崁入4毫升健康捐血者的血液中，然後以CellSearch系統進行處理。 Blood from healthy donors was aspirated into a vacuum blood collection tube containing EDTA, and then 4 ml was treated with the CellSearch system (Veridex, Raritan, NJ) to separate the CEC. This is the use of the CEC Spectrum Kit (Veridex), according to the manufacturer's guidelines. In the dowel experiment, 500 or 1000 endothelial cells (HAEC, HPAEC, HUVEC, or HMVEC-ad) were infiltrated into the blood of 4 ml healthy donors and then processed with the CellSearch system.

對培養過的內皮細胞或細胞團塊而言，約5×105個細胞係應用在採取AllPrep DNA/RNA微套件(德國希爾登的Qiagen公司)的RNA單離上。為了將RNA從CellSearch系統所濃縮後的CEC單離出來，先將350微升RLT plus緩衝液(Qiagen公司)加入裂解的CEC內，然後加入4微升、濃度為5奈克/微升的poly(I)(威斯康辛州麥迪遜的Epicentre公司)當作載體RNA，DNA和RNA係使用AllPrep DNA/RNA微套件、根據製造商指南加以單離。將2個相同的崁釘樣品匯集，以進行接下來的分析。RNA的定量和定性係使用NanoDrop 1000(德拉瓦州威爾明頓的NanoDrop公司)和Agilent Bioanalyzer 2100(加州聖克拉拉的Agilent公司)檢驗。 On cultured endothelial cells or cell aggregates, the approximately 5 × 10 5 cell lines studied taking AllPrep DNA / RNA Micro Kit (Qiagen, Hilden, Germany's) on the isolated RNA. In order to separate RNA from the CEC concentrated in the CellSearch system, 350 μl of RLT plus buffer (Qiagen) was added to the cleaved CEC, followed by 4 μl of a concentration of 5 Ng/μl of poly. (I) (Epicentre, Inc., Madison, Wis.) As a vector RNA, DNA and RNA were isolated using the AllPrep DNA/RNA micro-kit according to the manufacturer's guidelines. Two identical dowel samples were pooled for the next analysis. Quantification and characterization of RNA was performed using a NanoDrop 1000 (NanoDrop, Wilmington, Delaware) and an Agilent Bioanalyzer 2100 (Agilent, Inc., Santa Clara, Calif.).

內皮細胞RNA、平滑肌細胞RNA和PBMC RNA樣品係使用單股RNA放大和Enzo Biotin標示系統，轉換成標示目標反義RNA(cRNA)。目標物係依照供應商(加州聖克拉拉的Affymetrix公司)建議，雜合 到Affymetrix human U133 Plus 2.0陣列後續協議。雜合後，陣列係先使用標準Affymetrix程序進行清洗和染色，再使用Affymetrix GeneChip掃描儀進行掃描並使用Expression控制台進行數據萃取。 Endothelial RNA, smooth muscle cell RNA, and PBMC RNA samples were converted to the target antisense RNA (cRNA) using single-strand RNA amplification and the Enzo Biotin labeling system. Targets are recommended according to the supplier (Affymetrix, Santa Clara, Calif.) Follow-up protocol to the Affymetrix human U133 Plus 2.0 array. After hybridization, the array was first cleaned and stained using the standard Affymetrix program, scanned using an Affymetrix GeneChip scanner and data extracted using the Expression console.

對於CellSearch譜套件所處理的EC崁釘和捐贈血液樣品而言，RNA係使用Ovation RNA放大系統V2(加州聖卡洛斯的NuGEN公司)轉換成標示目標基因cDNA。簡言之，50奈克的總RNA係使用適於反轉錄的DNA/RNA嵌合引體，接著等溫放大來轉換成雙股cDNA。該cDNA係使用磁珠純化，並使用光譜法定量。3.75微克純化過的cDNA隨後歷經使用Encore Biotin模塊(NuGEN公司)所進行的兩步驟碎化和標示程序。首先，純化過的cDNA係進行碎化，以產生50至100個基範圍的單股cDNA產物。第二，這個碎化過的產物之標示，係將生物素標示過的核苷酸以酵素貼附至第一步驟中所生成之碎化過cDNA的3-羥基端。針對雜合，雜合雞尾酒係使用陣列所應用的雜合法、清洗和染色套件(Affymetrix公司)進行製備並將其加入碎化過的產物內，然後在45℃下培養18小時。雜合後，陣列係先使用標準Affymetrix程序進行清洗和染色，再使用Affymetrix GeneChip掃描儀進行掃描並使用Expression控制台進行數據萃取。 For EC nails and donated blood samples processed by the CellSearch Spectrum Suite, the RNA was converted to the target gene cDNA using the Ovation RNA Amplification System V2 (NuGEN Corporation, San Carlos, Calif.). Briefly, 50 ng of total RNA was converted to double-stranded cDNA using isozyme amplification using DNA/RNA chimeric primers suitable for reverse transcription. The cDNA was purified using magnetic beads and quantified using spectroscopy. 3.75 micrograms of purified cDNA was subsequently subjected to a two-step fragmentation and labeling procedure using the Encore Biotin module (NuGEN). First, the purified cDNA line is fragmented to produce a single-stranded cDNA product ranging from 50 to 100 bases. Second, the fragmented product is labeled by attaching the biotin-labeled nucleotide to the 3-hydroxyl end of the fragmented cDNA generated in the first step. For hybridization, the hybrid cocktail was prepared using the hybrid, wash and stain kit (Affymetrix) applied to the array and added to the shredded product, followed by incubation at 45 °C for 18 hours. After hybridization, the array was first cleaned and stained using the standard Affymetrix program, scanned using an Affymetrix GeneChip scanner and data extracted using the Expression console.

基因表現強度係使用MAS5演算法，以Affymetrix Expression控制台(1.1版)進行數據萃取。數據分析 前，先進行全球尺度化，以便將晶片的平均信號強度帶入目標值600。為了將雜訊程度降至最低，因此移除隊列中出現呼叫低於兩次的探針。因此，保留31K個探針組，以因應後續分析。觀察到從細胞培養取得的細胞和從細胞團塊取得的樣品間存有來源效應。為了將這種效應降至最低，因此移除在這2個族群間顯示出顯著差異(p<0.05)的探針。因此，獲得23K個探針。為了找到檢測血液中CEC適用的標記，因此移除2個PBMC樣品中出現呼叫一次或2個PBMC樣品任一者中強度超過200的探針，而保留3950個探針，以因應接下來的選擇。 Gene expression intensity was extracted using the MAS5 algorithm using the Affymetrix Expression console (version 1.1). data analysis Previously, global scaling was first performed to bring the average signal strength of the wafer to a target value of 600. To minimize the level of noise, remove probes that have fewer than two calls in the queue. Therefore, 31K probe sets were retained for subsequent analysis. A source effect was observed between cells taken from cell culture and samples taken from cell pellets. In order to minimize this effect, probes showing significant differences (p < 0.05) between the two ethnic groups were removed. Therefore, 23K probes were obtained. In order to find the marker for the detection of CEC in the blood, the probes with a strength of more than 200 in either of the 2 PBMC samples or 2 PBMC samples were removed, and 3950 probes were retained to accommodate the next selection. .

就層級式分群法而言，每個探針組的信號強度係規律化成中等；層級式分群法係使用Partek Genomics Suites(密蘇里州聖路易斯的Partek公司，6.5版)執行。層級式分群法係使用平均連結法，而執行在探針和樣品兩者上。歐幾里德距離係使用來計算相異性。就無監督層級式分群法而言，使用在移除細胞培養和細胞團塊間帶有顯著差異之基因後的23K個探針。監督的分群法係使用130個CEC標記或67個皮膚、動脈和靜脈內皮細胞特異標記加以執行。 In the case of hierarchical grouping, the signal intensity of each probe set is regularized to medium; the hierarchical grouping method is performed using Partek Genomics Suites (Partek, St. Louis, Miss., version 6.5). The hierarchical grouping method uses an average ligation method and is performed on both the probe and the sample. Euclidean distance is used to calculate dissimilarity. For the unsupervised hierarchical fractionation method, 23K probes after removal of genes with significant differences between cell culture and cell mass were used. The supervised clustering method was performed using 130 CEC markers or 67 skin, arterial and venous endothelial cell specific markers.

功能註釋係藉由使用DAVID軟體(NCIF公司)的Gene Ontology(GO公司)分類系統進行分析。 Functional annotations were analyzed by using the Gene Ontology (GO) classification system of DAVID Software (NCIF).

為了評估微陣列分析所選出的基因，因此從藉由使用CEC譜套件之CellSearch所處理的一組10個捐 贈樣品和8個細胞株崁釘樣品(2個HAEC崁釘、2個HPAEC崁釘、2個HMVEC崁釘和2個HMVEC-ad崁釘樣品)中萃取出RNA。在20微升反應中，使用2微升RNA進行使用高容量cDNA反轉錄套件(加州佛斯特市的Applied Biosystems公司)的cDNA合成。為了能夠進行多個基因分析，因此在20微升反應中，使用5微升cDNA進行使用TaqMan® PreAmp Master(前置放大核心)混合套件(Applied Biosystems公司)的14週期前置放大。前置放大產物係經1：20的稀釋，然後將5微升稀釋後產物使用來作為定量PCR的投入。即時PCR係使用Applied Biosystems基因表現Taqman檢定、在ABI PRISM 7900HT序列檢測儀(Applied Biosystems公司)上執行。 In order to evaluate the genes selected by the microarray analysis, a set of 10 donations processed by CellSearch using the CEC spectrum suite was used. RNA was extracted from the sample and 8 cell strain nail samples (2 HAEC nails, 2 HPAEC nails, 2 HMVEC nails, and 2 HMVEC-ad nail samples). In a 20 microliter reaction, 2 microliters of RNA was used for cDNA synthesis using a high capacity cDNA reverse transcription kit (Applied Biosystems, Inc., Foster, CA). In order to be able to perform multiple gene analyses, 14 microliters of cDNA was used for 14-cycle preamplification using a TaqMan® PreAmp Master (Applied Biosystems) mixing kit in a 20 microliter reaction. The preamplification product was diluted 1:20, and then 5 microliters of the diluted product was used as an input for quantitative PCR. The real-time PCR system was performed on an ABI PRISM 7900HT Sequence Detector (Applied Biosystems) using the Applied Biosystems gene expression Taqman assay.

實例2：基因表現分析 Example 2: Gene Expression Analysis

從18個內皮細胞樣品、2個PBMC樣品和2個平滑肌細胞樣品取得的RNA係歷經使用Affymetrix human U133 Plus 2.0陣列的微陣列分析，其中該陣列包含超過54000個探針組，涵蓋有著38,500個賦予良好特徵人類基因的47,000個轉錄和變種。內皮細胞樣品組代表9個不同解剖部位，包括5種不同血管(主動脈、冠狀動脈、肺動脈、髂動脈、臍動脈)、2種不同靜脈(臍靜脈和大隱靜脈)和2個不同組織(皮膚、心臟)。除髂動脈和皮膚之外，包括一個從培養細胞取 得和一個從冷凍細胞團塊取得的2個樣品係由各個來源取得。就髂動脈和皮膚而言，則使用從相同培養過細胞取得的2個樣品並作為技術副本。為了獲得基因表現模式概況，因此使用在至少2個樣品中顯示有出現呼叫的31K個探針組進行基因陣列數據的無監督層級分析。內皮細胞群的初步分析表明，從細胞培養取得的樣品和從細胞團塊取得的樣品，可能因樣品類型差異而形成不同族群。為了消除這種差異，因此在這兩個樣品群組間進行T檢驗。淘汰掉P值<0.05的探針組，結果為23K個探針組。在這些23K個探針組上的無監督分析產生出三個主要族群，分別為PBMC樣品族群、平滑肌細胞樣品族群和內皮細胞樣品族群，反映出內皮細胞與PBMC和平滑肌細胞相較之下、整體相似。在內皮細胞族群之內，從細胞團塊取得和從細胞培養取得的樣品係未分離，表明來源效應消除。然而，從相同解剖位置取得的樣品並非總是聚集在一起，可能是因為相對小的樣品尺寸和細胞培養不同條件所致。 RNAs obtained from 18 endothelial cell samples, 2 PBMC samples, and 2 smooth muscle cell samples were analyzed by microarray using an Affymetrix human U133 Plus 2.0 array, which contained more than 54,000 probe sets covering 38,500 grants Good characteristics of 47,000 transcriptions and variants of human genes. The endothelial cell sample group represents 9 different anatomical sites, including 5 different blood vessels (aorta, coronary artery, pulmonary artery, radial artery, umbilical artery), 2 different veins (umbilical vein and saphenous vein), and 2 different tissues ( Skin, heart). In addition to the brachial artery and skin, including one from the cultured cells Two samples taken from frozen cell pellets were obtained from various sources. For the brachial artery and skin, 2 samples taken from the same cultured cells were used as a technical copy. In order to obtain a profile of gene expression patterns, an unsupervised hierarchical analysis of gene array data was performed using 31K probe sets showing the presence of calls in at least 2 samples. Preliminary analysis of endothelial cell populations showed that samples taken from cell culture and samples taken from cell pellets may form different ethnic groups due to differences in sample types. To eliminate this difference, a T test was performed between the two sample groups. The probe set with a P value <0.05 was eliminated and the result was 23K probe sets. Unsupervised analysis on these 23K probe sets yielded three major populations, the PBMC sample population, the smooth muscle cell sample population, and the endothelial cell sample population, reflecting endothelial cells compared to PBMC and smooth muscle cells, overall. similar. Within the endothelial cell population, the sample taken from the cell mass and taken from the cell culture was not isolated, indicating that the source effect was eliminated. However, samples taken from the same anatomical location are not always clustered together, probably due to relatively small sample sizes and different conditions for cell culture.

實例3：CellSearch系統應用於CEC濃縮上。 Example 3: The CellSearch system was applied to CEC concentration.

CellSearch系統是用來將CEC從上述崁釘捐贈樣品中單離。對應於最突出內皮細胞膜抗原CD146的抗體係用於免疫擷取試劑。然而，在濃縮過程後，仍有約1000至5000個白血球殘留在濃縮後的CEC群體。 為了識別出潛在的CEC特異標記，內皮細胞中一個特異標記的表現程度必須實質高於其在PBMC中的表現程度。因此，移除2個PBMC樣品任一者中有出現呼叫一次或2個PBMC樣品任一者中強度超過200的探針組。保留3950個探針，以因應進一步分析。在18個樣品中最低強度低於1000的探針，係不考慮作為後續分析之用。因此，選出130個探針套(106個獨特基因)作為CEC標記的候選者。功能註釋和路徑分析係使用DAVID功能註釋軟體而在這些130個探針組上進行。GO(Gene ontology公司)術語和具有顯著過度代表的KEGG(京都基因和基因組百科全書)路徑如表1所示。在130個探針組中過度代表的頂端生物過程，包括參與細胞運動調節(n=11)、細胞遷移調節(n=10)和血管發育(n=11)的基因。該過度代表的細胞成分族群包括質膜(n=44)和集中附著點(n=6)。該過度代表的分子功能族群包括跨膜受體蛋白酪氨酸激酶活性(n=7)和蛋白酪氨酸激酶活性(n=8)。與CEC特異基因相關的主要路徑是集中附著點(n=12)和ECM受體相互作用(n=5)。內皮細胞其中一種功能是血管生成，在此期間細胞和細胞外基質(ECM)間的附著相互作用中發生重大變化，使得內皮細胞遷移。 The CellSearch system is used to separate CEC from the above donated samples. An anti-system corresponding to the most prominent endothelial cell membrane antigen CD146 was used for the immunological extraction reagent. However, after the concentration process, there are still about 1000 to 5000 white blood cells remaining in the concentrated CEC population. In order to identify potential CEC-specific markers, the extent of expression of a specific marker in endothelial cells must be substantially higher than that in PBMC. Therefore, one of the two PBMC samples was removed, and one of the probes or one of the two PBMC samples with a strength of more than 200 was present. 3950 probes were retained for further analysis. Probes with a minimum intensity below 1000 in 18 samples were not considered for subsequent analysis. Therefore, 130 probe sets (106 unique genes) were selected as candidates for CEC markers. Functional annotation and path analysis were performed on these 130 probe sets using the DAVID functional annotation software. The GO (Gene Onlogy) terminology and the KEGG (Kyoto Gene and Genomic Encyclopedia) path with significant over-representation are shown in Table 1. Top biological processes that are overrepresented in the 130 probe sets, including genes involved in cell motility regulation (n=11), cell migration regulation (n=10), and vascular development (n=11). The over-represented population of cellular components includes the plasma membrane (n=44) and the concentrated attachment point (n=6). This overrepresented molecular functional population includes transmembrane receptor protein tyrosine kinase activity (n=7) and protein tyrosine kinase activity (n=8). The major pathways associated with CEC-specific genes are concentrated attachment points (n=12) and ECM receptor interactions (n=5). One of the functions of endothelial cells is angiogenesis, during which significant changes occur in the attachment interaction between cells and the extracellular matrix (ECM), causing endothelial cells to migrate.

實例3：基因為基礎的標記 Example 3: Gene-based markers

CEC的特異基因係依照實例2進行識別。這些包括：整合相關基因，整合α2(I TGA2)、AXL受體酪氨酸激酶(AXL)、EPH受體A2(EPHA2)、TEK酪氨酸激酶、內皮(TEK)、原癌基因(MET)、神經纖毛蛋白1(NRP1)、VEGFR2(KDR)和TIE1；血管生成相關基因，激活素A受體類型II類1(ACVRL1)、結締組織生長因子(CTGF)、內皮細胞特異趨化調節器(ECSCR)、內皮素1(EDN1)和迴旋同源體4(ROBO4)；透過細胞-細胞相互作用而維持血管完整性的基因CAV1、CAV2、COL4A1、COL5A2、CCND1、FLNB、ITGA2、KDR、LAMA4、LAMB1、PARVA、MET；和獨立的MCAM(CD146)、KDR(VEGFR-2)、TEK(Tie-2)。然而，內皮細胞相關的一些基因並不是目前內文中的標記，因為其在PBMC中的高表現，如PECAM(CD31)、CXCR4，或因為其表現過低而在一個或多個內皮細胞株(例如KIT(SCF R/c-組)和SELE(E-選擇素))中無法見到對於診斷上的幫助。在這些實例中識別的CEC標記過度代表與內皮細胞功能相關的生物過程或路徑。 The specific gene line of CEC was identified according to Example 2. These include: integration of related genes, integration of α2 ( I TGA2 ), AXL receptor tyrosine kinase ( AXL ), EPH receptor A2 ( EPHA2 ), TEK tyrosine kinase, endothelial ( TEK ), proto-oncogene ( MET ) , neuropilin 1 ( NRP1 ), VEGFR2 ( KDR ) and TIE1 ; angiogenesis-related genes, activin A receptor type II 1 ( ACVRL1 ), connective tissue growth factor ( CTGF ), endothelial cell-specific chemotactic regulator ( ECSCR ), endothelin 1 ( EDN1 ) and convoluted homolog 4 ( ROBO4 ); genes CAV1, CAV2, COL4A1, COL5A2, CCND1, FLNB, ITGA2, KDR, LAMA4, which maintain vascular integrity through cell-cell interaction LAMB1, PARVA, MET; and independent MCAM (CD146), KDR (VEGFR-2), TEK (Tie-2). However, some genes associated with endothelial cells are not currently labeled in the context because of their high expression in PBMC, such as PECAM (CD31), CXCR4 , or because they are under-performing in one or more endothelial cell lines (eg Diagnostic assistance is not seen in KIT (SCF R/c-group) and SELE (E-selectin). The CEC markers identified in these examples are overrepresenting biological processes or pathways associated with endothelial cell function.

實例4：來源組織。 Example 4: Source organization.

在2個PBMC樣品中顯示沒有表現或強度小於200的3950個基因，其係用來識別有用於識別CEC來源組織的基因為基礎的標記。該等在從動脈取得之內皮細胞中之中位數信號強度超過500，且在從其他 來源取得之內皮細胞中大於最大表現者係識別為動脈內皮細胞特異基因。同樣，為了識別靜脈內皮細胞特異基因，識別其在從靜脈取得之內皮細胞中的中位數信號強度超過500，且在從其他來源取得之內皮細胞中大於最大表現之基因。共有38個動脈內皮細胞特異基因和14個靜脈內皮細胞特異基因符合這些標準(見表3)。代表性動脈特異基因是先前報告的動脈內皮細胞特異基因HEY2，其是已經暗示小鼠胚胎心血管發育所需之Hairy相關轉錄因子家族的成員。其他動脈特異CEC基因包括CXADR；其是一種緊密接合的成分且經報告在心臟中表現不對稱，其中表現係出現在血管壁的內皮下層中、但不出現在腔內皮細胞表面上。SOX17、一種HMG盒轉錄因子，其係已顯示在內胚層形成和心血管發育兩者中扮演重要角色，而其啟動子活性係已顯現在小鼠模型之心血管系統中的動脈血管內皮細胞內，但不顯現在靜脈內。為了啟動特別適合結合免疫磁性平台，如CellSearch系統使用之組織特異性標記的檢測，因此選定標記的表現程度應明顯高於在PBMC中者。就皮膚內皮細胞特異基因識別而言，則選擇從皮膚取得之內皮細胞中的最低表現，比從所有其他內皮細胞樣品之最高表現高出5倍的基因。此類的基因有15個。 3950 genes with no expression or intensity less than 200 were shown in 2 PBMC samples, which were used to identify genes based on genes used to identify CEC-derived tissues. These median signal intensities in endothelial cells obtained from arteries exceed 500, and those greater than the largest manifestations in endothelial cells obtained from other sources are recognized as arterial endothelial cell-specific genes. Similarly, in order to identify venous endothelial cell-specific genes, the median signal intensity in endothelial cells obtained from veins was identified to be more than 500, and the genes larger than the maximum expression in endothelial cells obtained from other sources. A total of 38 arterial endothelial cell-specific genes and 14 venous endothelial cell-specific genes met these criteria (see Table 3). A representative arterial-specific gene is the previously reported arterial endothelial cell-specific gene, HEY2 , which is a member of the Hairy-associated transcription factor family that has been implicated in the embryonic development of mouse embryos. Other arterial-specific CEC genes include CXADR ; it is a tightly joined component and has been reported to be asymmetrical in the heart, with manifestations occurring in the subendothelial layer of the vessel wall but not on the surface of the lumen endothelial cells. SOX17 , an HMG-box transcription factor, has been shown to play an important role in both endoderm formation and cardiovascular development, and its promoter activity has been shown in arterial vascular endothelial cells in the cardiovascular system of mouse models. But does not appear in the veins. In order to initiate detection of tissue-specific markers that are particularly suitable for use with immunomagnetic platforms, such as the CellSearch system, the degree of performance of selected markers should be significantly higher than in PBMC. For skin endothelial cell-specific gene recognition, the lowest performance in endothelial cells obtained from the skin was selected, which is five times higher than the highest performance of all other endothelial cell samples. There are 15 such genes.

監督分群法係使用67個來源特異基因，在細胞株微陣列數據上進行。該67個基因正確地聚集從不同來 源的內皮細胞，只有一個例外(一個動脈內皮細胞與靜脈內皮細胞聚集在一起)。亦進行基因的交叉驗證。 The supervised clustering method used 67 source-specific genes and performed on cell line microarray data. The 67 genes are correctly assembled from different There is only one exception to the source of endothelial cells (an arterial endothelial cell is clustered with venous endothelial cells). Cross-validation of genes was also performed.

實例5：利用內皮細胞崁釘進行標記驗證。 Example 5: Labeling validation using endothelial cell dowels.

內皮細胞係崁釘到健康捐贈血液中，然後CEC係使用CellSearch系統，搭配CEC譜套件進行濃縮。每4毫升健康捐贈血液係連同從單一來源取得的500或1000個培養過的內皮細胞進行崁釘。檢測4個選出的培養過的內皮細胞樣品，包括2個從動脈(HAEC和HPAEC)取得、1個從靜脈(HUVEC)取得、1個從皮膚(HMVEC-ad)取得。從崁釘取得的濃縮後CEC和無崁釘的10個健康捐贈樣品係歷經RNA單離和微陣列分析。從10個捐助者取得的CEC數值係確定在4毫升血液中為2至107。在從細胞株基因表現譜微陣列數據所獲得的130個CEC特異標記間，皆顯示在內皮細胞崁釘樣品中的平均表現高於捐贈樣品中者，在內皮細胞崁釘樣品中對捐贈樣品中之平均表現比率介於2到655，且93個基因的比例超過20。使用QRT-PCR，針對崁釘樣品中對捐贈樣品之間比率介於8到654的21個標記進行驗證，PCR檢測結果表明選出的所有標記在崁釘樣品和非崁釘捐贈樣品間皆有良好分離效果。為了驗證動脈、靜脈、皮膚特異性標記，使用67個來源特異標記針對崁釘微陣列數據進行主成分分析。捐贈樣品和動脈、靜脈、皮膚內皮細胞崁釘樣品 係在PCA中明顯地分離。該標記因此適用於區分從不同來源取得的CEC。 The endothelial cell system was nailed to healthy donor blood, and then the CEC system was condensed using the CellSearch system with a CEC spectrum kit. Each 4 ml of healthy donated blood line was nailed together with 500 or 1000 cultured endothelial cells obtained from a single source. Four selected cultured endothelial cell samples were examined, including two obtained from arteries (HAEC and HPAEC), one obtained from vein (HUVEC), and one obtained from skin (HMVEC-ad). Ten healthy donated samples of concentrated CEC and sputum-free nails obtained from dowels were subjected to RNA detachment and microarray analysis. The CEC values obtained from 10 donors were determined to be 2 to 107 in 4 ml of blood. Among the 130 CEC-specific markers obtained from the microarray data of the cell line gene expression profiles, the average performance in the endothelial cell nail samples was higher than that in the donor samples, and in the donor samples in the endothelial cell nail samples. The average performance ratio is between 2 and 655, and the proportion of 93 genes exceeds 20. Using QRT-PCR, 21 markers with a ratio between donated samples ranging from 8 to 654 were validated in the nail samples. The PCR results showed that all the selected markers were good between the nail samples and the non-nail nail samples. seperate effect. To verify arterial, venous, and skin-specific markers, principal component analysis was performed on the nail microarray data using 67 source-specific markers. Donated samples and arterial, venous, and skin endothelial cell nail samples The lines are clearly separated in the PCA. This marker is therefore suitable for distinguishing CECs obtained from different sources.

實例6：病情與CEC分析的關聯性(預知) Example 6: Correlation between disease and CEC analysis (predicted)

將從病患取得的全血直接放到含有CellSave防腐劑的試管內。此步驟係根據適於使用CellSave系統之循環腫瘤細胞(CTC)分析的相同收集血液協議進行。CEC係透過CellSearch系統，搭配CEC細胞擷取套件進行濃縮。在此系統中，首先搭配AUTOPREP分離系統進行免疫磁性濃縮，以便產生出濃縮後餾份。所使用的套件包含CD146磁性液體和利用核酸染料DAPI進行濃縮後細胞染色的試劑、內皮糖蛋白(CD105)-PE和泛-白血球標記CD45APC。將樣品減少到約300微升，並放置到固定在對應於磁單層細胞為內側之磁性「掩體」的分析室中。然後進行CEC的計數和形態分析。 Whole blood obtained from the patient was placed directly into a test tube containing CellSave preservative. This step was performed according to the same collected blood protocol suitable for circulating tumor cell (CTC) analysis using the CellSave system. The CEC is condensed with the CEC Cell Capture Kit via the CellSearch system. In this system, immunomagnetic concentration is first performed with the AUTOPREP separation system to produce a concentrated fraction. The kit used contained CD146 magnetic fluid and reagent for cell staining after concentration with the nucleic acid dye DAPI, endoglin (CD105)-PE and pan-white blood cell marker CD45APC. The sample was reduced to approximately 300 microliters and placed in an analysis chamber fixed to a magnetic "bunker" corresponding to the inner side of the magnetic monolayer. The CEC count and morphological analysis are then performed.

然後將CEC從磁性掩蔽設備中萃取出來。總RNA係利用RNeasy微套件(德國希爾登的Qiagen公司)進行萃取。該RNA係如下轉換成cDNA：首先，萃取過的RNA總量係在65℃下，以300奈克隨機六聚體預培養5分鐘。然後將200 U M-MLV Reverse Transcriptase(反轉錄酶)、RNase H Minus、Point Mutant、M-MLV Reverse Transcriptase 1x Reaction Buffer(反轉錄酶1倍反應緩衝液)、10 U RNasin® Plus RNase Inhibitor(抑制劑)(均購自威斯康星州麥迪遜的Promega公司)，50奈莫耳的dATP、dTTP、dCTP和dGTP等莫耳混合物(德國弗賴堡的Amersham Biosciences公司)和水添加至20微升的最後反應體積內。先在20℃下進行預培養步驟10分鐘後，然後在55℃下進行反應50分鐘。最後，加熱到94℃達5分鐘後，停止反應。 The CEC is then extracted from the magnetic masking device. Total RNA was extracted using the RNeasy micro-kit (Qiagen, Hilden, Germany). The RNA was converted to cDNA as follows: First, the total amount of extracted RNA was pre-incubated with a 300 ng random hexamer for 5 minutes at 65 °C. Then 200 U M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant, M-MLV Reverse Transcriptase 1x Reaction Buffer, 10 U RNasin® Plus RNase Inhibitor (all purchased from Promega, Madison, Wis.), 50 nm molar mixture of dATP, dTTP, dCTP and dGTP (Amersham Biosciences, Freiburg, Germany) and water added to 20 Microliters within the final reaction volume. The pre-incubation step was carried out at 20 ° C for 10 minutes and then at 55 ° C for 50 minutes. Finally, after heating to 94 ° C for 5 minutes, the reaction was stopped.

定量反轉錄PCR(qRTPCR)係然後如下執行。基因表現分析係使用表3中動脈標記指定的單獨TaqMan®預開發檢定試劑，進行重複反應。在每一種情況下，由2個未標示的PCR引體和1個搭配上述系統使用的FAMTM染料標示過TaqMan® MGB探針所組成。反應總體積為14 |xl，包含有7微升2倍的TaqMan® Universal PCR Master(通用PCR核心試劑)、0.7 |xl的TaqMan®預開發檢定試劑和4|xl 5倍稀釋的cDNA範本。PCR放大係使用AB 7900HT快速即時PCR系統進行，其由在50℃下初步培養2分鐘，然後在95℃下培養10分鐘、接著50個週期在95℃下變性15秒、然後在60℃下延伸1分鐘等步驟組成。利用AB7900序列檢測軟體2.2.2版，搭配自動基線校正和循環閾值設定進行數據分析。將所產生的循環閾值(Ct)數據匯出，以因應進一步分析。耗材、設備和軟體係購自加州福斯特城的Applied Biosystems公司。 Quantitative reverse transcription PCR (qRTPCR) was then performed as follows. Gene performance analysis was repeated using a separate TaqMan® pre-developed assay reagent designated by the arterial marker in Table 3. In each case, two unlabeled PCR primers and one FAMTM dye used in conjunction with the above system were labeled with a TaqMan® MGB probe. The total reaction volume was 14 |xl, containing 7 μl of 2-fold TaqMan® Universal PCR Master (general PCR core reagent), 0.7 μl of TaqMan® pre-developed assay reagent, and 4×xl 5-fold diluted cDNA template. PCR amplification was performed using the AB 7900HT Fast Real-Time PCR System, which was initially incubated at 50 °C for 2 minutes, then at 95 °C for 10 minutes, followed by 50 cycles at 95 °C for 15 seconds, then at 60 °C. 1 minute and other steps. Data analysis was performed using the AB7900 sequence detection software version 2.2.2 with automatic baseline correction and cycle threshold settings. The generated cycle threshold (Ct) data is remitted for further analysis. Consumables, equipment, and soft systems were purchased from Applied Biosystems, Inc., Foster City, California.

一些動脈CEC來源組織標記顯示出顯著的過度表現。將數據下載到一個程式使用的檔案，該程式會將該表現模式，與藉著將CEC標記表現與已知臨床結果匹配所執行的各種表現模式進行比較。該程式係包含在儲存設備上，而該儲存設備也包含使用統計算法進行比較適用的可執行碼。該模式表明CEC者本質上是動脈，更確切來說是主動脈。連同其他診斷訊息可結論出如果不及時治療，病患很可能招致胸主動脈動脈瘤。 Some arterial CEC-derived tissue markers show significant overexpression. The data is downloaded to a file used by the program, which compares the performance pattern to various performance patterns performed by matching CEC marker performance to known clinical outcomes. The program is included on the storage device, and the storage device also contains executable code that is more suitable for use with statistical algorithms. This pattern suggests that the CEC is essentially an artery, more specifically the aorta. Together with other diagnostic messages, it can be concluded that if left untreated, the patient is likely to have a thoracic aortic aneurysm.

>gi|86787911|ref|NM_004105.3|人類含EGF之fibulin樣細胞外間質蛋白1(EFEMP1)，轉錄 變體1，訊息核糖核酸 >gi|86787911|ref|NM_004105.3|human fibulin-like extracellular matrix protein 1 (EFEMP1), transcription Variant 1, message ribonucleic acid

>gi|13375818|ref|NM_024609.1|人類巢蛋白(NES)，訊息核糖核酸 >gi|13375818|ref|NM_024609.1|Human Nestin (NES), Message RNA

>ENA|BC025770|BC025770.1人類ras同源基因家族，成員J，訊息核糖核酸(互補型去氧核糖 核酸克隆影像：5210490)，****警告：嵌合克隆****。 >ENA|BC025770|BC025770.1 Human ras homologous gene family, member J, message ribonucleic acid (complementary deoxyribose) Nucleic acid clone image: 5210490), **** warning: chimeric clone ****.

>ENA|AK098525|AK098525.1人類互補型去氧核糖核酸FLJ25659 fis，克隆TST00427，高度 類似小家鼠刺？因子相互作用蛋白(Hip)訊息核糖核酸。 >ENA|AK098525|AK098525.1 Human Complementary Deoxyribonucleic Acid FLJ25659 fis, clone TST00427, height Similar to the little house mouse thorn? Factor interacting protein (Hip) message ribonucleic acid.

>ENA|U67280|U67280.1人類鈣腔蛋白訊息核糖核酸，完整編碼序列。 >ENA|U67280|U67280.1 Human calcium cavity protein message ribonucleic acid, complete coding sequence.

>ENA|AB032261|AB032261.1人類固醇輔？A去飽和？Scd訊息核糖核酸，完整編碼序列。 >ENA|AB032261|AB032261.1 Human sterol supplement? A desaturated? Scd message ribonucleic acid, complete coding sequence.

>refseq|NM_001554|NM_001554人類富含半胱胺酸，血管新生誘導因子，61(CYR61)，訊息 核糖核酸。 >refseq|NM_001554|NM_001554 Human-rich cysteine, angiogenesis-inducing factor, 61 (CYR61), message Ribonucleic acid.

>refseq|NM_001423|NM_001423人類上皮細胞膜蛋白1(EMP1)，訊息核糖核酸。 >refseq|NM_001423|NM_001423 Human epithelial membrane protein 1 (EMP1), message ribonucleic acid.

>refseq|NM_001387|NM_001387人類二氫嘧啶？樣3(DPYSL3)，轉錄變體2，訊息核糖核酸。 >refseq|NM_001387|NM_001387 Human dihydropyrimidine? Sample 3 (DPYSL3), transcript variant 2, message ribonucleic acid.

>refseq|NM_001839|NM_001839人類鈣調寧蛋白3，酸性(CNN3)，訊息核糖核酸。 >refseq|NM_001839|NM_001839 Human Calcine Protein 3, Acidic (CNN3), Message RNA.

>ENA|AI039874|AI039874.1 ox97c05.x1 Soares_senescent_fibroblasts_NbHSF人類互補型 去氧核糖核酸克隆影像：1664264 3'類似gb：J03934 NAD(P)H去氫？(人類)，訊息核糖核酸序 列。 >ENA|AI039874|AI039874.1 ox97c05.x1 Soares_senescent_fibroblasts_NbHSF Human Complementary Deoxyribonucleic acid cloning image: 1664264 3' similar to gb: J03934 NAD (P) H dehydrogenation? (human), message ribonucleotide Column.

>ENA|AL577531|AL577531.3人類胎盤COT 25態化CS0DI087YP20克隆之人類全長互補型去氧核 糖核酸3’端(人類) >ENA|AL577531|AL577531.3 Human placenta COT 25-state CS0DI087YP20 clone human full-length complementary deoxygen 3' end of the nucleic acid (human)

>refseq|NM_002933|NM_002933人類核糖核酸？，RNase A家族，1(胰)(RNASE1)，轉錄變體 4，訊息核糖核酸。 >refseq|NM_002933|NM_002933 Human ribonucleic acid? , RNase A family, 1 (pancreas) (RNASE1), transcript variant 4, message ribonucleic acid.

>ENA|AF079117|AF079117.1人類平衡NBMPR敏感核甘輸送載體(ENT1)訊息核糖核酸，完整編 碼序列。 >ENA|AF079117|AF079117.1 Human Balance NBMPR Sensitive Nuclear Delivery Vector (ENT1) Message RNA, Complete Edit Code sequence.

>refseq|NM_015577|NM_015577人類視黃酸誘導14(RAI14)，轉錄變體1，訊息核糖核酸。 >refseq|NM_015577|NM_015577 Human retinoic acid induced 14 (RAI14), transcript variant 1, message ribonucleic acid.

>refseq|NM_015472|NM_015472人類含WW區域轉錄調節物1(WWTR1)，轉錄變體1，訊息核糖 核酸。 >refseq|NM_015472|NM_015472 Human WW-containing transcriptional regulator 1 (WWTR1), transcript variant 1, message ribose Nucleic acid.

>refseq|NM_006169|NM_006169人類菸鹼胺氮甲基轉移？(NNMT)，訊息核糖核酸。 >refseq|NM_006169|NM_006169 Human Nicotinamide Nitrogen Methyl Transfer? (NNMT), message ribonucleic acid.

>refseq|NM_006169|NM_006169人類菸鹼胺氮甲基轉移？(NNMT)，訊息核糖核酸。 >refseq|NM_006169|NM_006169 Human Nicotinamide Nitrogen Methyl Transfer? (NNMT), message ribonucleic acid.

>ENA|AI754404|AI754404.1 cr24g06.x1人類骨髓基質細胞人類互補型去氧核糖核酸克隆 HBMSC_cr24g06 3'，訊息核糖核酸序列。 >ENA|AI754404|AI754404.1 cr24g06.x1 Human bone marrow stromal cells human complementary DNA clone HBMSC_cr24g06 3', message ribonucleic acid sequence.

>refseq|NM_000935|NM_000935人類原膠原離胺酸，2-氧戊二酸5-二氧？2(PLOD2)，轉錄變 體2，訊息核糖核酸。 >refseq|NM_000935|NM_000935 Human procollagen lysine, 2-oxoglutarate 5-diox? 2 (PLOD2), transcriptional change Body 2, message ribonucleic acid.

>refseq|NM_000602|NM_000602人類serpin胜？？抑制因子，分支E(nexin，第一類血纖維蛋 白溶？原激活物抑制因子)，成員1(SERPINE1)，轉錄變體1，訊息核糖核酸。 >refseq|NM_000602|NM_000602 Human serpin wins? ? Inhibitory factor, branch E (nexin, first type of fibrin egg White soluble? Pro-activator inhibitor), member 1 (SERPINE1), transcript variant 1, message ribonucleic acid.

>refseq|NM_021913|NM_021913人類AXL受體酪胺酸激脢(AXL)，轉錄變體1，訊息核糖核酸。 >refseq|NM_021913|NM_021913 Human AXL receptor tyrosine kinase (AXL), transcript variant 1, message ribonucleic acid.

>refseq|NM_004199|NM_004199人類脯胺醯-4-羥化？，α多？II(P4HA2)，轉錄變體1，訊息 核糖核酸。 >refseq|NM_004199|NM_004199 Human amidoxime-4-hydroxylation? , α more? II (P4HA2), transcript variant 1, message Ribonucleic acid.

>refseq|NM_000138|NM_000138人類原纖蛋白1(FBN1)，訊息核糖核酸。 >refseq|NM_000138|NM_000138 Human Fibrillin 1 (FBN1), message ribonucleic acid.

>refseq|NM_014899|NM_014899人類Rho相關BTB區域containing 3(RHOBTB3)，訊息核糖 核酸。 >refseq|NM_014899|NM_014899 Human Rho-related BTB area containing 3 (RHOBTB3), message ribose Nucleic acid.

>refseq|NM_002318|NM_002318人類lysyl oxidase樣2(LOXL2)，訊息核糖核酸。 >refseq|NM_002318|NM_002318 Human lysyl oxidase-like 2 (LOXL2), message ribonucleic acid.

>refseq|NM_016201|NM_016201人類angiomotin like 2(AMOTL2)，訊息核糖核酸。 >refseq|NM_016201|NM_016201 Human angiomotin like 2 (AMOTL2), message ribonucleic acid.

>ENA|BF197655|BF197655.1 7m95b04.x1 NCI_CGAP_Brn23人類互補型去氧核糖核酸克隆影 像：3562710 3'，訊息核糖核酸序列。 >ENA|BF197655|BF197655.1 7m95b04.x1 NCI_CGAP_Brn23 Human Complementary Deoxyribonucleic Acid Like: 3562710 3', message ribonucleic acid sequence.

>refseq|NM_001233|NM_001233人類caveolin 2(CAV2)，轉錄變體1，訊息核糖核酸。 >refseq|NM_001233|NM_001233 Human caveolin 2 (CAV2), transcript variant 1, message ribonucleic acid.

>refseq|NM_004431|NM_004431人類EPH受體A2(EPHA2)，訊息核糖核酸。 >refseq|NM_004431|NM_004431 Human EPH receptor A2 (EPHA2), message ribonucleic acid.

>ENA|BG170541|BG170541.1 602322942F1 NIH_MGC_89人類互補型去氧核糖核酸克隆影像： 4425947 5'，訊息核糖核酸序列。 >ENA|BG170541|BG170541.1 602322942F1 NIH_MGC_89 Human Complementary Deoxyribonucleic Acid Clone Image: 4425947 5', message ribonucleic acid sequence.

>refseq|NM_007034|NM_007034人類DnaJ(Hsp40)同源物，亞科B，成員4(DNAJB4)，訊息核 糖核酸。 >refseq|NM_007034|NM_007034 Human DnaJ (Hsp40) homolog, subfamily B, member 4 (DNAJB4), message core Sugar nucleic acid.

>refseq|NM_002253|NM_002253人類激？***區域受體(一第三類受體激酪胺酸激？)(KDR)， 訊息核糖核酸。 >refseq|NM_002253|NM_002253 Humans? Insertion region receptor (a third type of receptor tyrosine stimulator?) (KDR), Message RNA.

>refseq|NM_014890|NM_014890人類絲蛋白A結合蛋白1樣(FILIP1L)，轉錄變體2，訊息核糖 核酸。 >refseq|NM_014890|NM_014890 Human silk protein A binding protein 1 (FILIP1L), transcript variant 2, message ribose Nucleic acid.

>refseq|NM_002067|NM_002067人類鳥嘌呤核酸結合蛋白(G蛋白)，α11(Gq class) (GNA11)，訊息核糖核酸。 >refseq|NM_002067|NM_002067 Human guanine nucleic acid binding protein (G protein), α11 (Gq class) (GNA11), message ribonucleic acid.

>refseq|NM_003213|NM_003213人類TEA區域家族成員4(TEAD4)，轉錄變體1，訊息核糖核 酸。 >refseq|NM_003213|NM_003213 Human TEA region family member 4 (TEAD4), transcript variant 1, message ribonucleo acid.

>ENA|AL514445|AL514445.3人類神經母細胞瘤CL0BB010ZF08克隆之人類全長互補型去氧核糖 核酸3’端(人類) >ENA|AL514445|AL514445.3 Human neuroblastoma CL0BB010ZF08 cloned human full-length complementary deoxyribose Nucleic acid 3' end (human)

>refseq|NM_005613|NM_005613人類G-蛋白訊號傳遞4調節因子(RGS4)，轉錄變體2，訊息核 糖核酸。 >refseq|NM_005613|NM_005613 Human G-protein signal delivery 4 regulatory factor (RGS4), transcript variant 2, message core Sugar nucleic acid.

>ENA|BC000737|BC000737.1人類G-蛋白訊號傳遞4調節因子，訊息核糖核酸(互補型去氧核糖 核酸克隆MGC：2124影像：3510260)，完整編碼序列。 >ENA|BC000737|BC000737.1 Human G-protein signal delivery 4 regulatory factor, message ribonucleic acid (complementary deoxyribose Nucleic acid clone MGC: 2124 image: 3510260), complete coding sequence.

>refseq|NM_005424|NM_005424具免疫球蛋白樣區域及EGF樣區域1(TIE1)之人類激酪胺酸 激？，訊息核糖核酸。 >refseq|NM_005424|NM_005424 Human tyrosine with immunoglobulin-like region and EGF-like region 1 (TIE1) Excited? , message ribonucleic acid.

>ENA|BE962749|BE962749.2 601656143R1 NIH_MGC_66人類互補型去氧核糖核酸克隆影像： 3855754 3'，訊息核糖核酸序列。 >ENA|BE962749|BE962749.2 601656143R1 NIH_MGC_66 Human Complementary Deoxyribonucleic Acid Clone Image: 3855754 3', message ribonucleic acid sequence.

>refseq|NM_012242|NM_012242人類dickkopf同源物1(Xenopus laevis)(DKK1)，訊息核 糖核酸。 >refseq|NM_012242|NM_012242 Human Dickkopf homolog 1 (Xenopus laevis) (DKK1), message core Sugar nucleic acid.

>refseq|NM_001424|NM_001424人類上皮細胞膜蛋白2(EMP2)，訊息核糖核酸。 >refseq|NM_001424|NM_001424 Human epithelial membrane protein 2 (EMP2), message ribonucleic acid.

>ENA|U29586|U29586.1人類β-肌膜蛋白聚糖肌肉萎縮蛋白相關醣蛋白訊息核糖核酸，完整 編碼序列。 >ENA|U29586|U29586.1 Human beta-myosin dystrophin-associated glycoprotein message ribonucleic acid, complete Coding sequence.

>refseq|NM_015976|NM_015976人類分選蛋白7(SNX7)，轉錄變體1，訊息核糖核酸。 >refseq|NM_015976|NM_015976 Human Sorting Protein 7 (SNX7), Transcriptional Variant 1, Message RNA.

>refseq|NM_000950|NM_000950人類富含脯胺酸Gla(G-羧基麩胺酸)1(PRRG1)，轉錄變體1 ，訊息核糖核酸。 >refseq|NM_000950|NM_000950 Human rich in proline Gla (G-carboxy glutamic acid) 1 (PRRG1), transcript variant 1 , message ribonucleic acid.

>refseq|NM_005795|NM_005795人類抑鈣素受體樣(CALCRL)，訊息核糖核酸。 >refseq|NM_005795|NM_005795 Human calcitonin receptor-like (CALCRL), message ribonucleic acid.

>refseq|NM_000459|NM_000459人類TEK激酪胺酸激？，內皮(TEK)，訊息核糖核酸。 >refseq|NM_000459|NM_000459 Human TEK tyrosine acid stimulation? , endothelium (TEK), message ribonucleic acid.

>refseq|NM_003662|NM_003662人類pirin(鐵離子結合核蛋白)(PIR)，轉錄變體1，訊息核 糖核酸。 >refseq|NM_003662|NM_003662 Human pirin (iron ion binding nucleoprotein) (PIR), transcript variant 1, message nucleus Sugar nucleic acid.

>refseq|NM_001235|NM_001235人類絲胺酸蛋白？抑制劑胜？？抑制因子，分支H(熱休克蛋白 47)，成員1，(膠原結合蛋白1)(SERPINH1)，轉錄變體2，訊息核糖核酸。 >refseq|NM_001235|NM_001235 Human serine protein? Inhibitor wins? ? Inhibitory factor, branch H (heat shock protein) 47), member 1, (collagen binding protein 1) (SERPINH1), transcript variant 2, message ribonucleic acid.

>refseq|NM_003483|NM_003483人類高遷移性族群AT-hook 2(HMGA2)，轉錄變體1，訊息核 糖核酸。 >refseq|NM_003483|NM_003483 human high mobility group AT-hook 2 (HMGA2), transcript variant 1, message core Sugar nucleic acid.

>ENA|AV712733|AV712733.1人類互補型去氧核糖核酸克隆：DCAACE01，5'端，表達於人類樹 突細胞中。 >ENA|AV712733|AV712733.1 Human Complementary Deoxyribonucleic Acid Clone: DCAACE01, 5' End, Expressed in Human Tree In the cell.

>ENA|M73554|M73554.1人類bcl-1訊息核糖核酸，完整編碼序列。 >ENA|M73554|M73554.1 Human bcl-1 message ribonucleic acid, complete coding sequence.

>ENA|BC004295|BC004295.1人類，克隆影像：3622356，訊息核糖核酸，部份編碼序列。 >ENA|BC004295|BC004295.1 Human, clone image: 3622356, message ribonucleic acid, partial coding sequence.

>ENA|AF312393|AF312393.1人類白胺酸拉鏈蛋白FKSG13(FKSG13)訊息核糖核酸，完整編碼 序列。 >ENA|AF312393|AF312393.1 Human leucine zipper protein FKSG13 (FKSG13) message ribonucleic acid, complete coding sequence.

>ENA|AL078459|AL078459.8來自染色體1p11.4-21.3上RP4-621F18克隆之人類DNA序列，包 含一部分新穎基因，及二甲基精胺酸二甲基胺基水解？1之DDAH1基因3'端。 >ENA|AL078459|AL078459.8 Human DNA sequence from RP4-621F18 clone on chromosome 1p11.4-21.3, package Contains a part of the novel gene, and the hydrolysis of dimethyl arginine dimethylamine? 1 DDAH1 gene 3' end.

>ENA|M92934|M92934.1人類結締組織生長因子訊息核糖核酸，完整編碼序列。 >ENA|M92934|M92934.1 Human connective tissue growth factor message ribonucleic acid, complete coding sequence.

>ENA|U84895|U84895.1人類跨膜4總科A15同源物訊息核糖核酸，完整編碼序列。 >ENA|U84895|U84895.1 Human transmembrane 4 superfamily A15 homolog message ribonucleic acid, complete coding sequence.

>ENA|AL037401|AL037401.1人類訊息核糖核酸；EST DKFZp564K1671_s1(來自克隆 DKFZp564K1671) >ENA|AL037401|AL037401.1 Human Message RNA; EST DKFZp564K1671_s1 (from clone DKFZp564K1671)

>ENA|M90657|M90657.1人類腫瘤抗原(L6)訊息核糖核酸，完整編碼序列。 >ENA|M90657|M90657.1 Human tumor antigen (L6) message ribonucleic acid, complete coding sequence.

>ENA|D84109|D84109.1人類RBP-MS/第三類訊息核糖核酸，完整編碼序列。 >ENA|D84109|D84109.1 Human RBP-MS/Group III message ribonucleic acid, complete coding sequence.

>ENA|D84109|D84109.1人類RBP-MS/第三類訊息核糖核酸，完整編碼序列。 >ENA|D84109|D84109.1 Human RBP-MS/Group III message ribonucleic acid, complete coding sequence.

>ENA|J03225|J03225.1人類脂蛋白關聯凝結抑制因子訊息核糖核酸，完整編碼序列。 >ENA|J03225|J03225.1 Human Lipoprotein Associated Condensation Inhibitor Information RNA, Complete Coding Sequence

>ENA|BC001258|BC001258.1人類葡萄糖磷酸變位？3，訊息核糖核酸(互補型去氧核糖核酸克 隆MGC：5002影像：3451677)，完整編碼序列。 >ENA|BC001258|BC001258.1 Human Glucose Phosphorylation? 3, message ribonucleic acid (complementary DNA gram Long MGC: 5002 image: 3451677), complete coding sequence.

>ENA|BC004241|BC004241.1人類層黏蛋白，α4，訊息核糖核酸(互補型去氧核糖核酸克隆 影像：3623597)，完整編碼序列。 >ENA|BC004241|BC004241.1 Human laminin, α4, message ribonucleic acid (complementary DNA clone) Image: 3623597), complete coding sequence.

>ENA|AF026219|AF026219.1人類HP蛋白(HP)訊息核糖核酸，完整編碼序列。 >ENA|AF026219|AF026219.1 Human HP protein (HP) message ribonucleic acid, complete coding sequence.

>ENA|AF003114|AF003114.1人類CYR61訊息核糖核酸，完整編碼序列。 >ENA|AF003114|AF003114.1 Human CYR61 message ribonucleic acid, complete coding sequence.

>ENA|U17473|U17473.1人類抑鈣素樣受器訊息核糖核酸，完整編碼序列。 >ENA|U17473|U17473.1 Human calcitonin-like receptor message ribonucleic acid, complete coding sequence.

>ENA|BC004908|BC004908.1人類互補型去氧核糖核酸克隆影像：3530459，****警告：嵌合 克隆****。 >ENA|BC004908|BC004908.1 Human Complementary Deoxyribonucleic Acid Clone Image: 3530459, **** Warning: Chimeric clone****.

>ENA|M28882|M28882.1人類MUC18醣蛋白訊息核糖核酸，完整編碼序列。 >ENA|M28882|M28882.1 Human MUC18 glycoprotein message ribonucleic acid, complete coding sequence.

>ENA|BC003096|BC003096.1人類PDZ及LIM區域4，訊息核糖核酸(互補型去氧核糖核酸克隆 MGC：1645影像：3501794)，完整編碼序列。 >ENA|BC003096|BC003096.1 Human PDZ and LIM Region 4, Message Ribonucleic Acid (Complementary DNA Cloning MGC: 1645 image: 3501794), complete coding sequence.

>ENA|M20206|M20206.1人類層黏蛋白B1訊息核糖核酸，完整編碼序列。 >ENA|M20206|M20206.1 Human laminin B1 message ribonucleic acid, complete coding sequence.

>ENA|AI922605|AI922605.1 wm90c05.x1 NCI_CGAP_Ut2人類互補型去氧核糖核酸克隆影像： 2443208 3'，訊息核糖核酸序列。 >ENA|AI922605|AI922605.1 wm90c05.x1 NCI_CGAP_Ut2 Human Complementary Deoxyribonucleic Acid Clone Image: 2443208 3', message ribonucleic acid sequence.

>ENA|AI695017|AI695017.1 we45d07.x1 NCI_CGAP_Lu24人類互補型去氧核糖核酸克隆影 像：2344045 3'，訊息核糖核酸序列。 >ENA|AI695017|AI695017.1 we45d07.x1 NCI_CGAP_Lu24 Human Complementary Deoxyribonucleic Acid Like: 2340445 3', message ribonucleic acid sequence.

>ENA|BE552421|BE552421.1 hw26b02.x1 NCI_CGAP_Kid11人類互補型去氧核糖核酸克隆影 像：3184011 3'類似TR：094872 094872 KIAA0774蛋白，訊息核糖核酸序列。 >ENA|BE552421|BE552421.1 hw26b02.x1 NCI_CGAP_Kid11 Human Complementary Deoxyribonucleic Acid Like: 3184011 3' Similar to TR: 094872 094872 KIAA0774 protein, message ribonucleic acid sequence.

>ENA|AU147399|AU147399.1人類互補型去氧核糖核酸克隆：MAMMA1000563，3'端，表達於乳腺。 >ENA|AU147399|AU147399.1 Human complementary DNA clone: MAMMA1000563, 3' end, expressed in the mammary gland.

>ENA|N95026|N95026.1 zb45d12.s1 Soares_fetal_lung_NbHL19W人類互補型去氧核糖核酸 克隆影像：306551 3'類似包含元素MSR1反覆元素，訊息核糖核酸序列。 >ENA|N95026|N95026.1 zb45d12.s1 Soares_fetal_lung_NbHL19W Human Complementary Deoxyribonucleic Acid Clone image: 306551 3' similar to the element MSR1 repeat element, message ribonucleic acid sequence.

>ENA|BE620457|BE620457.1 601483690F1 NIH_MGC_69人類互補型去氧核糖核酸克隆影像： 3886055 5'，訊息核糖核酸序列。 >ENA|BE620457|BE620457.1 601483690F1 NIH_MGC_69 Human Complementary Deoxyribonucleic Acid Clone Image: 3886055 5', message ribonucleic acid sequence.

>ENA|BF115739|BF115739.1 7n64b08.x1 NCI_CGAP_Lu24人類互補型去氧核糖核酸克隆影 像：3569246 3'，訊息核糖核酸序列。 >ENA|BF115739|BF115739.1 7n64b08.x1 NCI_CGAP_Lu24 Human Complementary Deoxyribonucleic Acid Like: 3569246 3', message ribonucleic acid sequence.

>ENA|AI935123|AI935123.1 wp13h09.x1 NCI_CGAP_Kid12人類互補型去氧核糖核酸克隆影 像：2464769 3'，訊息核糖核酸序列。 >ENA|AI935123|AI935123.1 wp13h09.x1 NCI_CGAP_Kid12 Human Complementary Deoxyribonucleic Acid Like: 2464769 3', message ribonucleic acid sequence.

>ENA|AI088622|AI088622.1 qb14f06.x1 Soares_pregnant_uterus_NbHPU人類互補型去氧核 糖核酸克隆影像：1696259 3'類似gb：X06409 RAF原致癌基因絲胺酸/蘇胺酸-蛋白激？(人 類)；包含PTR5.t3 TAR1反覆元素，訊息核糖核酸序列。 >ENA|AI088622|AI088622.1 qb14f06.x1 Soares_pregnant_uterus_NbHPU Human Complementary Deoxygen Glyconucleotide clone image: 1696259 3' similar gb: X06409 RAF proto-oncogene serine/threonine-protein activation? (people Class); contains PTR5.t3 TAR1 repetitive element, message ribonucleic acid sequence.

>ENA|AA917899|AA917899.1 o176e05.s1 NCI_CGAP_Kid3人類互補型去氧核糖核酸克隆影 像：1535552 3'，訊息核糖核酸序列。 >ENA|AA917899|AA917899.1 o176e05.s1 NCI_CGAP_Kid3 Human Complementary Deoxyribonucleic Acid Like: 1535552 3', message ribonucleic acid sequence.

>ENA|AW149379|AW149379.1 xf36d11.x1 NCI_CGAP_Brn50人類互補型去氧核糖核酸克隆影 像：2620149 3'類似TR：043251 043251未知功能39.5 KD蛋白，訊息核糖核酸序列。 >ENA|AW149379|AW149379.1 xf36d11.x1 NCI_CGAP_Brn50 Human Complementary Deoxyribonucleic Acid Like: 2620149 3' Similar to TR: 043251 043251 Unknown function 39.5 KD protein, message ribonucleic acid sequence.

>ENA|AW129021|AW129021.1 xe93h03.x1 NCI_CGAP_Brn35人類互補型去氧核糖核酸克隆影 像：2615477 3'類似TR：095034 095034 WUGSC：H_RG041D11.1蛋白，訊息核糖核酸序列。 >ENA|AW129021|AW129021.1 xe93h03.x1 NCI_CGAP_Brn35 Human Complementary Deoxyribonucleic Acid Like: 2615477 3' Similar to TR: 095034 095034 WUGSC: H_RG041D11.1 protein, message ribonucleic acid sequence.

>refseq|NM_018212|NM_018212人類致能化同源物(果蠅)(ENAH)，轉錄變體2，訊息核糖核 酸。 >refseq|NM_018212|NM_018212 Human-enhanced homolog (Drosophila) (ENAH), transcript variant 2, message ribonucleo acid.

>refseq|NM_018222|NM_018222人類parvin，α(PARVA)，訊息核糖核酸。 >refseq|NM_018222|NM_018222 Human parvin, alpha (PARVA), message ribonucleic acid.

>refseq|NM_012193|NM_012193人類卷曲家族受體4(FZD4)，訊息核糖核酸。 >refseq|NM_012193|NM_012193 Human Curl Family Receptor 4 (FZD4), message ribonucleic acid.

>refseq|NM_017734|NM_017734人類palmdelphin(PALMD)，訊息核糖核酸。 >refseq|NM_017734|NM_017734 Human palmdelphin (PALMD), message ribonucleic acid.

>refseq|NM_001955|NM_001955人類端內皮素1(EDN1)，轉錄變體1，訊息核糖核酸。 >refseq|NM_001955|NM_001955 Human Endothelin 1 (EDN1), Transcriptional Variant 1, Message RNA.

>refseq|NM_014344|NM_014344人類四連接框1(果蠅)(FJX1)，訊息核糖核酸。 >refseq|NM_014344|NM_014344 Human four junction box 1 (Drosophila) (FJX1), message ribonucleic acid.

>refseq|NM_017805|NM_017805人類Ras結合蛋白1(RASIP1)，訊息核糖核酸。 >refseq|NM_017805|NM_017805 Human Ras Binding Protein 1 (RASIP1), message ribonucleic acid.

>refseq|NM_000393|NM_000393人類膠原，第五類，α2(COL5A2)，訊息核糖核酸。 >refseq|NM_000393|NM_000393 Human collagen, fifth class, α2 (COL5A2), message ribonucleic acid.

>ENA|AK025108|AK025108.1人類互補型去氧核糖核酸：FLJ21455 fis，克隆COL04696. >ENA|AK025108|AK025108.1 Human Complementary Deoxyribonucleic Acid: FLJ21455 fis, clone COL04696.

>ENA|BG107577|BG107577.1 602277726F1 NIH_MGC_86人類互補型去氧核糖核酸克隆影像： 4365271 5'，訊息核糖核酸序列。 >ENA|BG107577|BG107577.1 602277726F1 NIH_MGC_86 Human Complementary Deoxyribonucleic Acid Clone Image: 4365271 5', message ribonucleic acid sequence.

>ENA|AF322916|AF322916.1人類葡萄膜自動抗原訊息核糖核酸，完整編碼序列。 >ENA|AF322916|AF322916.1 Human uveal autoantigen message ribonucleic acid, complete coding sequence.

>ENA|AF278532|AF278532.1人類β-導蛋白訊息核糖核酸，完整編碼序列。 >ENA|AF278532|AF278532.1 Human beta-conductin message ribonucleic acid, complete coding sequence.

>ENA|AY009951|AY009951.1人類刺？因子相互作用蛋白訊息核糖核酸，完整編碼序列。 >ENA|AY009951|AY009951.1 Human thorn? Factor interacting protein message ribonucleic acid, complete coding sequence.

>ENA|AA524250|AA524250.1 ng34a11.s1 NCI_CGAP_Co3人類互補型去氧核糖核酸克隆影像： 936668 3'，訊息核糖核酸序列。 >ENA|AA524250|AA524250.1 ng34a11.s1 NCI_CGAP_Co3 Human Complementary Deoxyribonucleic Acid Clone Image: 936668 3', message ribonucleic acid sequence.

>ENA|BF247906|BF247906.1 601858274F1 NIH_MGC_58人類互補型去氧核糖核酸克隆影像： 4068810 5'，訊息核糖核酸序列。 >ENA|BF247906|BF247906.1 601858274F1 NIH_MGC_58 Human Complementary Deoxyribonucleic Acid Clone Image: 4068810 5', message ribonucleic acid sequence.

>ENA|BG285417|BG285417.1 602409786F1 NIH_MGC_91人類互補型去氧核糖核酸克隆影像： 4539428 5'，訊息核糖核酸序列。 >ENA|BG285417|BG285417.1 602409786F1 NIH_MGC_91 Human Complementary Deoxyribonucleic Acid Clone Image: 4539428 5', message ribonucleic acid sequence.

>ENA|BF000162|BF000162.1 7h18f09.x1 NCI_CGAP_Co16人類互補型去氧核糖核酸克隆影 像：3316361 3'，訊息核糖核酸序列。 >ENA|BF000162|BF000162.1 7h18f09.x1 NCI_CGAP_Co16 Human Complementary Deoxyribonucleic Acid Like: 3316361 3', message ribonucleic acid sequence.

>ENA|N30138|N30138.1 yx81c07.s1 Soares黑素細胞2NbHM人類互補型去氧核糖核酸克隆影 像：268140 3'，訊息核糖核酸序列。 >ENA|N30138|N30138.1 yx81c07.s1 Soares melanocyte 2NbHM human complementary DNA clone Like: 268140 3', message ribonucleic acid sequence.

>ENA|AL040051|AL040051.2人類訊息核糖核酸；EST DKFZp434P1112_s1(來自克隆 DKFZp434P1112) >ENA|AL040051|AL040051.2 Human Message RNA; EST DKFZp434P1112_s1 (from clone DKFZp434P1112)

>ENA|AA156022|AA156022.1 zo48c05.s1 Stratagene內皮細胞937223人類互補型去氧核糖核 酸克隆影像：590120 3'，訊息核糖核酸序列。 >ENA|AA156022|AA156022.1 zo48c05.s1 Stratagene endothelial cells 937223 human complementary deoxyribose nucleus Acid clone image: 590120 3', message ribonucleic acid sequence.

>ENA|AA554833|AA554833.1 ni34g07.s1 NCI_CGAP_Lu1人類互補型去氧核糖核酸克隆影像： 978780 3'，訊息核糖核酸序列。 >ENA|AA554833|AA554833.1 ni34g07.s1 NCI_CGAP_Lu1 Human Complementary Deoxyribonucleic Acid Clone Image: 978780 3', message ribonucleic acid sequence.

>ENA|BG290908|BG290908.1 602387068F1 NIH_MGC_93人類互補型去氧核糖核酸克隆影像： 4515905 5'，訊息核糖核酸序列。 >ENA|BG290908|BG290908.1 602387068F1 NIH_MGC_93 Human Complementary Deoxyribonucleic Acid Clone Image: 4515905 5', message ribonucleic acid sequence.

>ENA|AW193693|AW193693.1 xm29d12.x1 NCI_CGAP_GC6人類互補型去氧核糖核酸克隆影像： 2685623 3'，訊息核糖核酸序列。 >ENA|AW193693|AW193693.1 xm29d12.x1 NCI_CGAP_GC6 Human Complementary Deoxyribonucleic Acid Clone Image: 2685623 3', message ribonucleic acid sequence.

>ENA|T63524|T63524.1 yc07b04.s1 Stratagene肺(#937210)人類互補型去氧核糖核酸克隆 影像：79951 3'，訊息核糖核酸序列。 >ENA|T63524|T63524.1 yc07b04.s1 Stratagene Lung (#937210) Human Complementary Deoxyribonucleic Acid Clone Image: 79951 3', message ribonucleic acid sequence.

>ENA|N95414|N95414.1 zb66d10.s1 Soares_fetal_lung_NbHL19W人類互補型去氧核糖核酸 克隆影像：308563 3'，訊息核糖核酸序列。 >ENA|N95414|N95414.1 zb66d10.s1 Soares_fetal_lung_NbHL19W Human Complementary Deoxyribonucleic Acid Clone image: 308563 3', message ribonucleic acid sequence.

>ENA|BF511276|BF511276.1 UI-H-BI4-aoj-d-05-0-UI.s1 NCI_CGAP_Sub8人類互補型去氧核 糖核酸克隆影像：3085089 3'，訊息核糖核酸序列。 >ENA|BF511276|BF511276.1 UI-H-BI4-aoj-d-05-0-UI.s1 NCI_CGAP_Sub8 Human Complementary Deoxygen Glyconucleotide clone image: 3085089 3', message ribonucleic acid sequence.

>ENA|AL571557|AL571557.3人類胎盤COT 25常態化CS0DI024YP08克隆之人類全長互補型去 氧核糖核酸3’端(人類) >ENA|AL571557|AL571557.3 Human placenta COT 25 normalized CS0DI024YP08 clone human full complement complement 3' end of ribonucleic acid (human)

>ENA|AA173223|AA173223.1 zp31b04.s1 Stratagene神經上皮(#937231)人類互補型去氧核 糖核酸克隆影像：611023 3類似包含Alu反覆元素；，訊息核糖核酸序列。 >ENA|AA173223|AA173223.1 zp31b04.s1 Stratagene Neuroepithelial (#937231) Human Complementary Deoxygen Glyconucleotide clone image: 611023 3 similarly contains Alu repetitive elements;, message ribonucleic acid sequence.

>ENA|AI623211|AI623211.1 ts78e11.x1 NCI_CGAP_GC6人類互補型去氧核糖核酸克隆影像 2237420 3'類似gb：M33552淋巴細胞特定蛋白LSP1(人類)，訊息核糖核酸序列。 >ENA|AI623211|AI623211.1 ts78e11.x1 NCI_CGAP_GC6 Human Complementary Deoxyribonucleic Acid Clone Image 2237420 3' Similar to gb: M33552 lymphocyte-specific protein LSP1 (human), message ribonucleic acid sequence.

>ENA|AI807004|AI807004.1 wf37a06.x1 Soares_NFL_T_GBC_S1人類互補型去氧核糖核酸克 隆影像：2357746 3'，訊息核糖核酸序列。 >ENA|AI807004|AI807004.1 wf37a06.x1 Soares_NFL_T_GBC_S1 Human Complementary Deoxyribonucleic Acid Image: 2357746 3', message ribonucleic acid sequence.

>ENA|AA181256|AA181256.1 zp58c07.s1 Stratagene內皮細胞937223人類互補型去氧核糖核 酸克隆影像：624396 3'，訊息核糖核酸序列。 >ENA|AA181256|AA181256.1 zp58c07.s1 Stratagene endothelial cells 937223 human complementary deoxyribose nucleus Acid clone image: 624396 3', message ribonucleic acid sequence.

>ENA|AI653117|AI653117.1 wb43b10.x1 NCI_CGAP_GC6人類互補型去氧核糖核酸克隆影像： 2308411 3'，訊息核糖核酸序列。 >ENA|AI653117|AI653117.1 wb43b10.x1 NCI_CGAP_GC6 Human Complementary Deoxyribonucleic Acid Clone Image: 2308411 3', message ribonucleic acid sequence.

>ENA|BE566894|BE566894.1 601341011F1 NIH_MGC_53人類互補型去氧核糖核酸克隆影像： 3683306 5'，訊息核糖核酸序列。 >ENA|BE566894|BE566894.1 601341011F1 NIH_MGC_53 Human Complementary Deoxyribonucleic Acid Clone Image: 3683306 5', message ribonucleic acid sequence.

>ENA|AI670852|AI670852.1 wa06a05.x1 NCI_CGAP_Kid11人類互補型去氧核糖核酸克隆影 像：2297264 3'，訊息核糖核酸序列。 >ENA|AI670852|AI670852.1 wa06a05.x1 NCI_CGAP_Kid11 Human Complementary Deoxyribonucleic Acid Like: 2297264 3', message ribonucleic acid sequence.

>ENA|AL035086|AL035086.12人類DNA序列來自6q23.1-24.3染色體上之RP1-44A20克隆， 包含新穎蛋白類似C1-四氫葉酸綜合體(包含DKFZP586G1517蛋白及FLJ21145，包含四氫葉酸 脫氫？/環化水解？區域)部分基因、部分新穎基因(包含FLJ31738及KIAA1209)及一新穎偽基 因類似光導蛋白樣蛋白。包含CpG島。 >ENA|AL035086|AL035086.12 Human DNA sequence from RP1-44A20 clone on chromosome 6q23.1-24.3, Contains a novel protein similar to the C1-tetrahydrofolate complex (comprising DKFZP586G1517 protein and FLJ21145, containing tetrahydrofolate Dehydrogenation? / cyclization hydrolysis? Regional) partial genes, some novel genes (including FLJ31738 and KIAA1209) and a novel pseudo-base Because it is similar to a photoprotein-like protein. Contains CpG islands.

>ENA|AI657069|AI657069.1 tt55g01.x1 NCI_CGAP_GC6人類互補型去氧核糖核酸克隆影像： 2244720 3'類似SW：KLP1_CHLRE P46870運動素樣蛋白KLP1，訊息核糖核酸序列。 >ENA|AI657069|AI657069.1 tt55g01.x1 NCI_CGAP_GC6 Human Complementary Deoxyribonucleic Acid Clone Image: 2244720 3' Similar to SW: KLP1_CHLRE P46870 kinetin-like protein KLP1, message ribonucleic acid sequence.

>ENA|AL135787|AL135787.13來自染色體9上RP11-16L21克隆之人類DNA序列，包含KIAA1962 蛋白類似鋅指蛋白HIT-10基因變化之3'端、白三烯B412-羥基去氫？(MGC34943)之LTB4DH基 因、KIAA0563相關基因(LOC286331)之基因、鳥嘌呤核酸結合蛋白(G蛋白)之GNG10基因、γ 10、新穎基因、染色體9開放讀框84之C90rf84基因之3'端及六個CpG島。 >ENA|AL135787|AL135787.13 Human DNA sequence from RP11-16L21 clone on chromosome 9, including KIAA1962 The protein is similar to the 3' end of the zinc finger protein HIT-10 gene change, and the leukotriene B412-hydroxy dehydrogenation? (MGC34943) LTB4DH base Gene, KIAA0563 related gene (LOC286331) gene, guanine nucleic acid binding protein (G protein) GNG10 gene, γ 10. Novel gene, chromosome 3 open reading frame 84 of the 3' end of the C90rf84 gene and six CpG islands.

>ENA|BG540685|BG540685.1 602570561F1 NIH_MGC_77人類互補型去氧核糖核酸克隆影像： 4694894 5'，訊息核糖核酸序列。 >ENA|BG540685|BG540685.1 602570561F1 NIH_MGC_77 Human Complementary Deoxyribonucleic Acid Clone Image: 4694894 5', message ribonucleic acid sequence.

>ENA|AW960748|AW960748.1 EST372819 MAGE重排序列，MAGF人類互補型去氧核糖核酸，訊 息核糖核酸序列。 >ENA|AW960748|AW960748.1 EST372819 MAGE Rearrangement Sequence, MAGF Human Complementary Deoxyribonucleic Acid, The ribonucleic acid sequence.

>ENA|AI583530|AI583530.1 ts12c08.x1 NCI_CGAP_Pan1人類互補型去氧核糖核酸克隆影 像：2228366 3'，訊息核糖核酸序列。 >ENA|AI583530|AI583530.1 ts12c08.x1 NCI_CGAP_Pan1 Human Complementary Deoxyribonucleic Acid Like: 2228366 3', message ribonucleic acid sequence.

>ENA|BF792126|BF792126.1 602252573F1 NIH_MGC_84人類互補型去氧核糖核酸克隆影像： 4344858 5'，訊息核糖核酸序列。 >ENA|BF792126|BF792126.1 602252573F1 NIH_MGC_84 Human Complementary Deoxyribonucleic Acid Clone Image: 4344858 5', message ribonucleic acid sequence.

>ENA|AW014647|AW014647.1 UI-H-BI0p-abd-b-12-0-UI.s1 NCI_CGAP_Sub2人類互補型去氧 核糖核酸克隆影像：2711375 3'，訊息核糖核酸序列。 >ENA|AW014647|AW014647.1 UI-H-BI0p-abd-b-12-0-UI.s1 NCI_CGAP_Sub2 Human Complementary Deoxygenation Ribonucleic acid cloning image: 2711375 3', message ribonucleic acid sequence.

>ENA|AW444502|AW444502.1 UI-H-BI3-akb-g-03-0-UI.s1 NCI_CGAP_Sub5人類互補型去氧核 糖核酸克隆影像：2733869 3'，訊息核糖核酸序列。 >ENA|AW444502|AW444502.1 UI-H-BI3-akb-g-03-0-UI.s1 NCI_CGAP_Sub5 Human Complementary Deoxygen Glyconucleotide clone image: 2733869 3', message ribonucleic acid sequence.

>ENA|BE218803|BE218803.1 hv44h08.x1 NCI_CGAP_Lu24人類互補型去氧核糖核酸克隆影 像：3176319 3'類似SW：RTC0_人類P17081 GTP結合蛋白TC10，訊息核糖核酸序列。 >ENA|BE218803|BE218803.1 hv44h08.x1 NCI_CGAP_Lu24 Human Complementary Deoxyribonucleic Acid Like: 3176319 3' Similar SW: RTC0_ human P17081 GTP binding protein TC10, message ribonucleic acid sequence.

>ENA|BE218803|BE218803.1 hv44h08.x1 NCI_CGAP_Lu24人類互補型去氧核糖核酸克隆影 像：3176319 3'類似SW：RTC0_人類P17081 GTP結合蛋白TC10，訊息核糖核酸序列。 >ENA|BE218803|BE218803.1 hv44h08.x1 NCI_CGAP_Lu24 Human Complementary Deoxyribonucleic Acid Like: 3176319 3' Similar SW: RTC0_ human P17081 GTP binding protein TC10, message ribonucleic acid sequence.

>ENA|AI628689|AI628689.1 ty43a06.x1 NCI_CGAP_Ut2人類互補型去氧核糖核酸克隆影像： 2281810 3'類似包含Alu反覆元素；包含元素MER22反覆元素，訊息核糖核酸序列。 >ENA|AI628689|AI628689.1 ty43a06.x1 NCI_CGAP_Ut2 Human Complementary Deoxyribonucleic Acid Clone Image: 2281810 3' similarly contains Alu repetitive elements; contains element MER22 repetitive elements, message ribonucleic acid sequence.

>ENA|W72516|W72516.1 zd64g05.s1 Soares_fetal_heart_NbHH19W人類互補型去氧核糖核酸 克隆影像：345464 3'，訊息核糖核酸序列。 >ENA|W72516|W72516.1 zd64g05.s1 Soares_fetal_heart_NbHH19W Human Complementary Deoxyribonucleic Acid Clone image: 345464 3', message ribonucleic acid sequence.

>refseq|NM_001387|NM_001387人類二氫嘧啶？樣3(DPYSL3)，轉錄變體2，訊息核糖核酸。 >refseq|NM_001387|NM_001387 Human dihydropyrimidine? Sample 3 (DPYSL3), transcript variant 2, message ribonucleic acid.

>refseq|NM_000943|NM_000943人類脯胺基異構？C(cyclophilin C)(PPIC)，訊息核糖核 酸。 >refseq|NM_000943|NM_000943 Human amidinoisomer? C (cyclophilin C) (PPIC), message ribonucleoside acid.

>refseq|NM_002841|NM_002841人類蛋白質酪胺酸去磷酸？，受體型，G(PTPRG)，訊息核糖 核酸。 >refseq|NM_002841|NM_002841 Human protein tyrosine dephosphorylation? , receptor type, G (PTPRG), message ribose Nucleic acid.

>refseq|NM_013409|NM_013409人類濾泡抑素(FST)，轉錄變體FST344，訊息核糖核酸。 >refseq|NM_013409|NM_013409 Human follicle inhibitor (FST), transcript variant FST344, message ribonucleic acid.

>refseq|NM_006207|NM_006207人類血小板衍生生長因子受體樣(PDGFRL)，訊息核糖核酸。 >refseq|NM_006207|NM_006207 Human platelet-derived growth factor receptor-like (PDGFRL), message ribonucleic acid.

>refseq|NM_004791|NM_004791人類***素，β樣1(具EGF樣重複區域)(ITGBL1)，訊息核 糖核酸。 >refseq|NM_004791|NM_004791 human insert, beta-like 1 (with EGF-like repeat region) (ITGBL1), message core Sugar nucleic acid.

>refseq|NM_004186|NM_004186人類sema區域，免疫球蛋白區域(Ig)，短鹼性區域，分泌 型，(semaphorin)3F(SEMA3F)，訊息核糖核酸。 >refseq|NM_004186|NM_004186 human sema region, immunoglobulin region (Ig), short alkaline region, secretory Type, (semaphorin) 3F (SEMA3F), message ribonucleic acid.

>ENA|U38276|U38276.1人類腦信號蛋白III家族同源訊息核糖核酸，完整編碼序列。 >ENA|U38276|U38276.1 Human brain signaling protein III family homologous message ribonucleic acid, complete coding sequence.

>ENA|AF055585|AF055585.1人類神經胞外裂隙蛋白Slit2訊息核糖核酸，完整編碼序列。 >ENA|AF055585|AF055585.1 Human neuronal extracellular fissure protein Slit2 message ribonucleic acid, complete coding sequence.

>ENA|AF056085|AF056085.1人類GABA-B受體訊息核糖核酸，完整編碼序列。 >ENA|AF056085|AF056085.1 Human GABA-B receptor message ribonucleic acid, complete coding sequence.

>ENA|AF095784|AF095784.1人類GABA-B受體R2(GABBR2)訊息核糖核酸，完整編碼序列。 >ENA|AF095784|AF095784.1 Human GABA-B receptor R2 (GABBR2) message ribonucleic acid, complete coding sequence.

>ENA|AL359052|AL359052.1人類訊息核糖核酸全長***互補型去氧核糖核酸克隆EUROIMAGE 1968422。 >ENA|AL359052|AL359052.1 Human message ribonucleic acid full-length insertion complementary DNA clone EUROIMAGE 1968422.

>ENA|AF095723|AF095723.nu112002年2月15日停止公開檢視。 >ENA|AF095723|AF095723.nu11 The public inspection was stopped on February 15, 2002.

N N

>refseq|NM_012193|NM_012193人類卷曲家族受體4(FZD4)，訊息核糖核酸。 >refseq|NM_012193|NM_012193 Human Curl Family Receptor 4 (FZD4), message ribonucleic acid.

>refseq|NM_021939|NM_021939人類FK506結合蛋白10，65 kDa(FKBP10)，訊息核糖核酸。 >refseq|NM_021939|NM_021939 Human FK506 binding protein 10,65 kDa (FKBP10), message ribonucleic acid.

>refseq|NM_012259|NM_012259與YRPW模體2(HEY2)相關人類髮狀/***增強子，訊息核糖核 酸。 >refseq|NM_012259|NM_012259 is associated with YRPW motif 2 (HEY2) human hair/split enhancer, message ribonucleo acid.

>refseq|NM_022454|NM_022454人類SRY(性別覺定區Y)-框17(SOX17)，訊息核糖核酸。 >refseq|NM_022454|NM_022454 Human SRY (Gender Sensing Zone Y) - Box 17 (SOX17), Message RNA.

>ENA|AK023795|AK023795.1人類互補型去氧核糖核酸FLJ13733 fis，克隆PLACE3000147， 高度類似於具第一類凝血？敏感蛋白ADAMTS1模體(ADAMTS1)人類金屬蛋白？訊息核糖核酸。 >ENA|AK023795|AK023795.1 Human Complementary Deoxyribonucleic Acid FLJ13733 fis, clone PLACE3000147, Highly similar to the first type of coagulation? Sensitive protein ADAMTS1 motif (ADAMTS1) human metalloprotein? Message RNA.

>ENA|AF060152|AF060152.1人類METH1蛋白(METH1)訊息核糖核酸，完整編碼序列。 >ENA|AF060152|AF060152.1 Human METH1 protein (METH1) message ribonucleic acid, complete coding sequence.

>ENA|AF232238|AF232238.1人類HES相關阻遏蛋白1HERP1訊息核糖核酸，完整編碼序列。 >ENA|AF232238|AF232238.1 Human HES-associated repressor protein 1 HERP1 message ribonucleic acid, complete coding sequence.

>ENA|AI049973|AI049973.1 an38g03.x1 Gessler Wilms腫瘤人類互補型去氧核糖核酸克隆 影像：1700980 3'，訊息核糖核酸序列。 >ENA|AI049973|AI049973.1 an38g03.x1 Gessler Wilms tumor human complementary DNA clone Image: 1700980 3', message ribonucleic acid sequence.

>ENA|BG260087|BG260087.1 602371673F1 NIH_MGC_93人類互補型去氧核糖核酸克隆影像： 4479642 5'，訊息核糖核酸序列。 >ENA|BG260087|BG260087.1 602371673F1 NIH_MGC_93 Human Complementary Deoxyribonucleic Acid Clone Image: 4479642 5', message ribonucleic acid sequence.

>ENA|BF438173|BF438173.1 7q68a03.x1 NCI_CGAP_Lu24人類互補型去氧核糖核酸克隆影 像：3703205 3'，訊息核糖核酸序列。 >ENA|BF438173|BF438173.1 7q68a03.x1 NCI_CGAP_Lu24 Human Complementary Deoxyribonucleic Acid Like: 3703205 3', message ribonucleic acid sequence.

>ENA|H16409|H16409.1 ym22h04.s1 Soares嬰兒大腦1NIB人類互補型去氧核糖核酸克隆影 像：49055 3'，訊息核糖核酸序列。 >ENA|H16409|H16409.1 ym22h04.s1 Soares baby brain 1NIB human complementary DNA clone Like: 49055 3', message ribonucleic acid sequence.

>ENA|BF338870|BF338870.1 602036126F1 NCI_CGAP_Brn64人類互補型去氧核糖核酸克隆影 像：4183999 5'，訊息核糖核酸序列。 >ENA|BF338870|BF338870.1 602036126F1 NCI_CGAP_Brn64 Human Complementary Deoxyribonucleic Acid Like: 4183999 5', message ribonucleic acid sequence.

>ENA|AI742043|AI742043.1 wg38c09.x1 Soares_NSF_F8_9W_OT_PA_P_S1人類互補型去氧核 糖核酸克隆影像：2367376 3'，訊息核糖核酸序列。 >ENA|AI742043|AI742043.1 wg38c09.x1 Soares_NSF_F8_9W_OT_PA_P_S1 Human Complementary Deoxygen Glyconucleotide clone image: 2367376 3', message ribonucleic acid sequence.

>ENA|BE644809|BE644809.1 7e57e09.x1 Soares_NSF_F8_9W_OT_PA_P_S1人類互補型去氧核 糖核酸克隆影像：3286600 3'類似包含元素MER37反覆元素，訊息核糖核酸序列。 >ENA|BE644809|BE644809.1 7e57e09.x1 Soares_NSF_F8_9W_OT_PA_P_S1 Human Complementary Deoxygen Glyconucleotide clone image: 3286600 3' similar to the element MER37 repeat element, message ribonucleic acid sequence.

>ENA|AI963304|AI963304.1 wt61d01.x1 NCI_CGAP_Pan1人類互補型去氧核糖核酸克隆影 像：2511937 3'類似gb：J03464 PROCOLLAGEN A2(I)鏈前驅(人類)，訊息核糖核酸序列。 >ENA|AI963304|AI963304.1 wt61d01.x1 NCI_CGAP_Pan1 Human Complementary Deoxyribonucleic Acid Like: 2511937 3' Similar to gb: J03464 PROCOLLAGEN A2 (I) chain precursor (human), message ribonucleic acid sequence.

>ENA|AW006182|AW006182.1 wz93c05.x1 NCI_CGAP_Brn25人類互補型去氧核糖核酸克隆影 像：2566376 3'，訊息核糖核酸序列。 >ENA|AW006182|AW006182.1 wz93c05.x1 NCI_CGAP_Brn25 Human Complementary Deoxyribonucleic Acid Like: 2566376 3', message ribonucleic acid sequence.

>ENA|AB037820|AB037820.1人類訊息核糖核酸for KIAA1399蛋白，部份編碼序列。 >ENA|AB037820|AB037820.1 Human Message RNA for KIAA1399 Protein, Partially Encoded Sequence.

>ENA|AI912122|AI912122.1 wd63a12.x1 NCI_CGAP_Lu24人類互補型去氧核糖核酸克隆影 像：2336254 3'，訊息核糖核酸序列。 >ENA|AI912122|AI912122.1 wd63a12.x1 NCI_CGAP_Lu24 Human Complementary Deoxyribonucleic Acid Like: 2336254 3', message ribonucleic acid sequence.

>ENA|AI376433|AI376433.1 tc36a08.x1 Soares_total_fetus_Nb2HF8_9w人類互補型去氧核 糖核酸克隆影像：2066678 3'，訊息核糖核酸序列。 >ENA|AI376433|AI376433.1 tc36a08.x1 Soares_total_fetus_Nb2HF8_9w Human Complementary Deoxygen Glyconucleotide clone image: 2066678 3', message ribonucleic acid sequence.

>ENA|AW960748|AW960748.1 EST372819 MAGE重排序列，MAGF人類互補型去氧核糖核酸，訊 息核糖核酸序列。 >ENA|AW960748|AW960748.1 EST372819 MAGE Rearrangement Sequence, MAGF Human Complementary Deoxyribonucleic Acid, The ribonucleic acid sequence.

>ENA|AW058459|AW058459.1 wx21b12.x1 NCI_CGAP_Kid11人類互補型去氧核糖核酸克隆影 像：2544287 3'，訊息核糖核酸序列。 >ENA|AW058459|AW058459.1 wx21b12.x1 NCI_CGAP_Kid11 Human Complementary Deoxyribonucleic Acid Like: 2544287 3', message ribonucleic acid sequence.

>ENA|AA904430|AA904430.1 ok07f12.s1 Soares_NFL_T_GBC_S1人類互補型去氧核糖核酸克 隆影像：1507151 3'類似TR：000808 Q00808 B轉導蛋白樣蛋白，訊息核糖核酸序列。 >ENA|AA904430|AA904430.1 ok07f12.s1 Soares_NFL_T_GBC_S1 Human Complementary Deoxyribonucleic Acid Image: 1507151 3' Similar to TR: 000808 Q00808 B transduction protein-like protein, message ribonucleic acid sequence.

>ENA|AF493879|AF493879.1人類鳥嘌呤核酸結合蛋白γ12(GNG12)訊息核糖核酸，完整編 碼序列。 >ENA|AF493879|AF493879.1 Human guanine nucleic acid binding protein γ12 (GNG12) message ribonucleic acid, complete editing Code sequence.

>ENA|AI753143|AI753143.1 cr05h01.x1人類骨髓基質細胞人類互補型去氧核糖核酸克隆 HBMSC_cr05h01 3'，訊息核糖核酸序列。 >ENA|AI753143|AI753143.1 cr05h01.x1 Human bone marrow stromal cells human complementary DNA clone HBMSC_cr05h01 3', message ribonucleic acid sequence.

>refseq|NM_015577|NM_015577人類視黃酸誘導14(RAI14)，轉錄變體1，訊息核糖核酸。 >refseq|NM_015577|NM_015577 Human retinoic acid induced 14 (RAI14), transcript variant 1, message ribonucleic acid.

>refseq|NM_005045|NM_005045人類搖晃素(RELN)，轉錄變體1，訊息核糖核酸。 >refseq|NM_005045|NM_005045 Human Shake White (RELN), Transcriptional Variant 1, Message RNA.

>ENA|AL037401|AL037401.1人類訊息核糖核酸；EST DKFZp564K1671_s1(來自 DKFZp564K1671克隆) >ENA|AL037401|AL037401.1 Human Message RNA; EST DKFZp564K1671_s1 (from DKFZp564K1671 clone)

>ENA|BC004241|BC004241.1人類層黏蛋白，α4，訊息核糖核酸(互補型去氧核糖核酸克隆 影像：3623597)，完整編碼序列。 >ENA|BC004241|BC004241.1 Human laminin, α4, message ribonucleic acid (complementary DNA clone) Image: 3623597), complete coding sequence.

>ENA|U77706|U77706.1人類層黏蛋白α4鏈(LAMA4*-1)訊息核糖核酸，完整編碼序列。 >ENA|U77706|U77706.1 Human laminin α4 chain (LAMA4*-1) message ribonucleic acid, complete coding sequence.

>ENA|U56725|U56725.1人類熱休克蛋白訊息核糖核酸，完整編碼序列。 >ENA|U56725|U56725.1 Human heat shock protein message ribonucleic acid, complete coding sequence.

>refseq|NM_018214|NM_018214人類富含白胺酸重複序列1(LRRC1)，訊息核糖核酸。 >refseq|NM_018214|NM_018214 Human is rich in leucine repeat 1 (LRRC1), message ribonucleic acid.

>refseq|NM_005092|NM_005092人類腫瘤壞死因子(配體)總科，成員18(TNFSF18)，訊息核 糖核酸。 >refseq|NM_005092|NM_005092 Human Tumor Necrosis Factor (Ligangology), Family Member 18 (TNFSF18), Message Nucleus Sugar nucleic acid.

>ENA|AI279819|AI279819.1 qm26h04.x1 NCI_CGAP_Lu5人類互補型去氧核糖核酸克隆影像： 1882999 3'，訊息核糖核酸序列。 >ENA|AI279819|AI279819.1 qm26h04.x1 NCI_CGAP_Lu5 Human Complementary Deoxyribonucleic Acid Clone Image: 1882999 3', message ribonucleic acid sequence.

>ENA|BE857360|BE857360.1 7g29b12.x1 NCI_CGAP_Brn23人類互補型去氧核糖核酸克隆影 像：3307871 3'，訊息核糖核酸序列。 >ENA|BE857360|BE857360.1 7g29b12.x1 NCI_CGAP_Brn23 Human Complementary Deoxyribonucleic Acid Like: 3307871 3', message ribonucleic acid sequence.

>ENA|AI801973|AI801973.1 tx29d05.x1 NCI_CGAP_Lu24人類互補型去氧核糖核酸克隆影 像：2270985 3'，訊息核糖核酸序列。 >ENA|AI801973|AI801973.1 tx29d05.x1 NCI_CGAP_Lu24 Human Complementary Deoxyribonucleic Acid Like: 2270985 3', message ribonucleic acid sequence.

>ENA|AW451115|AW451115.1 UI-H-BI3-alg-g-01-0-UI.s1 NCI_CGAP_Sub5人類互補型去氧核 糖核酸克隆影像：2736936 3'，訊息核糖核酸序列。 >ENA|AW451115|AW451115.1 UI-H-BI3-alg-g-01-0-UI.s1 NCI_CGAP_Sub5 Human Complementary Deoxygen Glyconucleotide clone image: 2736936 3', message ribonucleic acid sequence.

>ENA|AI472310|AI472310.1 tj87b01.x1 Soares_NSF_F8_9W_OT_PA_P_S1人類互補型去氧核 糖核酸克隆影像：2148457 3'，訊息核糖核酸序列。 >ENA|AI472310|AI472310.1 tj87b01.x1 Soares_NSF_F8_9W_OT_PA_P_S1 Human Complementary Deoxygen Glyconucleotide clone image: 2148457 3', message ribonucleic acid sequence.

>gi|18089116|gb|BC020718.1|人類補體因子I，訊息核糖核酸(互補型去氧核糖核酸克隆 MGC：22501影像：4716122)，完整編碼序列 >gi|18089116|gb|BC020718.1|Human complement factor I, message ribonucleic acid (complementary DNA clone) MGC: 22501 image: 4716122), complete coding sequence

>ENA|BF967657|BF967657.1 602287358T1 NIH_MGC_96人類互補型去氧核糖核酸克隆影像： 4374495 3'，訊息核糖核酸序列。 >ENA|BF967657|BF967657.1 602287358T1 NIH_MGC_96 Human Complementary Deoxyribonucleic Acid Clone Image: 4374495 3', message ribonucleic acid sequence.

>refseq|NM_006474|NM_006474人類腎小球足突細胞黏蛋白(PDPN)，轉錄變體1，訊息核糖核 酸。 >refseq|NM_006474|NM_006474 Human glomerular podocyte cell mucin (PDPN), transcript variant 1, message ribonucleo acid.

>refseq|NM_003619|NM_003619人類蛋白？，絲胺酸，12(神經胰蛋白？、馬達蛋白) (PRSS12)，訊息核糖核酸。 >refseq|NM_003619|NM_003619Human protein? , serine, 12 (neurotrypin?, motor protein) (PRSS12), message ribonucleic acid.

>refseq|NM_003149|NM_003149人類SH3及富含半胱氨酸區域(STAC)，訊息核糖核酸。 >refseq|NM_003149|NM_003149 Human SH3 and cysteine-rich region (STAC), message ribonucleic acid.

>ENA|AW007532|AW007532.1 ws52h07.x1 NCI_CGAP_Brn25人類互補型去氧核糖核酸克隆影 像：2500861 3'，訊息核糖核酸序列。 >ENA|AW007532|AW007532.1 ws52h07.x1 NCI_CGAP_Brn25 Human Complementary Deoxyribonucleic Acid Like: 2500861 3', message ribonucleic acid sequence.

>ENA|AI810767|AI810767.1 tu04d02.x1 NCI_CGAP_Pr28人類互補型去氧核糖核酸克隆影 像：2250051 3'，訊息核糖核酸序列。 >ENA|AI810767|AI810767.1 tu04d02.x1 NCI_CGAP_Pr28 Human Complementary Deoxyribonucleic Acid Like: 2250051 3', message ribonucleic acid sequence.

>ENA|AF154054|AF154054.1人類DRM(DRM)訊息核糖核酸，完整編碼序列。 >ENA|AF154054|AF154054.1 Human DRM (DRM) message ribonucleic acid, complete coding sequence.

>refseq|NM_013372|NM_013372人類gremlin 1(GREM1)，轉錄變體1，訊息核糖核酸。 >refseq|NM_013372|NM_013372 Human gremlin 1 (GREM1), transcript variant 1, message ribonucleic acid.

>ENA|AU154455|AU154455.1人類互補型去氧核糖核酸克隆：NT2RP4001145，3'端，導入視黃 酸(RA)兩週後表達於NT2神經前驅細胞中。 >ENA|AU154455|AU154455.1 Human Complementary Deoxyribonucleic Acid Clone: NT2RP4001145, 3' End, Introducing Retinal Acid (RA) was expressed in NT2 neural precursor cells two weeks later.

>ENA|AW590196|AW590196.1 hg41a02.x1 NCI_CGAP_GC6人類互補型去氧核糖核酸克隆影像： 2948138 3'，訊息核糖核酸序列。 >ENA|AW590196|AW590196.1 hg41a02.x1 NCI_CGAP_GC6 Human Complementary Deoxyribonucleic Acid Clone Image: 2948138 3', message ribonucleic acid sequence.

>ENA|BG494007|BG494007.1 602542289F1 NIH_MGC_59人類互補型去氧核糖核酸克隆影像： 4673182 5'，訊息核糖核酸序列。 >ENA|BG494007|BG494007.1 602542289F1 NIH_MGC_59 Human Complementary Deoxyribonucleic Acid Clone Image: 4673182 5', message ribonucleic acid sequence.

>ENA|AI540210|AI540210.1 te55e07.x1 Soares_NFL_T_GBC_S1人類互補型去氧核糖核酸克 隆影像：2090628 3'類似TR：Q63571 Q63571鼠生長與轉形依靠，訊息核糖核酸序列。 >ENA|AI540210|AI540210.1 te55e07.x1 Soares_NFL_T_GBC_S1 Human Complementary Deoxyribonucleic Acid Image: 2090628 3' Similar to TR: Q63571 Q63571 Rat growth and transformation depends on the message ribonucleic acid sequence.

>ENA|BF589515|BF589515.1 naa05c06.x1 NCI_CGAP_Pr28人類互補型去氧核糖核酸克隆影 像：3254002 3'，訊息核糖核酸序列。 >ENA|BF589515|BF589515.1 naa05c06.x1 NCI_CGAP_Pr28 Human Complementary Deoxyribonucleic Acid Like: 3254002 3', message ribonucleic acid sequence.

>gi|11444320|gb|BF432206.1|BF432206 nab81e11.x1 Soares_NSF_F8_9W_OT_PA_P_S1人類 互補型去氧核糖核酸克隆影像：3274101 3'，訊息核糖核酸序列 >gi|11444320|gb|BF432206.1|BF432206 nab81e11.x1 Soares_NSF_F8_9W_OT_PA_P_S1 human Complementary DNA cloning image: 3274101 3', message ribonucleic acid sequence

>gi|283806665|ref|NM_153448.3|人類ESX同源框1(ESX1)，訊息核糖核酸 >gi|283806665|ref|NM_153448.3|Human ESX homeobox 1 (ESX1), message ribonucleic acid

Claims (20)

一種評估醫療狀況的方法，包含識別CEC中基因為基礎的標記之不同表現(相對於一個正常群體中之相同基因的表現)。 A method of assessing a medical condition comprising identifying different manifestations of a gene-based marker in a CEC (relative to the performance of the same gene in a normal population). 如申請專利範圍第1項的方法，其中該標記在PBMC中表現並無不同。 The method of claim 1, wherein the label does not differ in performance in the PBMC. 如申請專利範圍第1項的方法，其中在該調控基因中的表現至少有2倍差異。 The method of claim 1, wherein the performance in the regulatory gene is at least 2-fold different. 如申請專利範圍第1項的方法，其中表示不同調控的p-值低於0.05。 The method of claim 1, wherein the p-value of the different regulation is less than 0.05. 如申請專利範圍第1項的方法，其中該標記係從表2所選出。 The method of claim 1, wherein the label is selected from Table 2. 一種識別一CEC來源組織的方法，包含識別CEC中之組織特異基因為基礎的標記不同表現(相對於在一個正常群體中之相同基因的表現)。 A method of identifying a CEC-derived tissue comprising identifying different expressions of a marker based on a tissue-specific gene in CEC (relative to the performance of the same gene in a normal population). 如申請專利範圍第6項的方法，其中該標記在PBMC中表現並無不同。 The method of claim 6, wherein the label does not differ in performance in the PBMC. 如申請專利範圍第1項的方法，其中在該調控基因中的表現至少有2倍差異。 The method of claim 1, wherein the performance in the regulatory gene is at least 2-fold different. 如申請專利範圍第1項方法，其中表示不同調控的p-值低於0.05。 For example, the method of claim 1 of the patent scope, wherein the p-value of different regulation is less than 0.05. 如申請專利範圍第1項的方法，其中該標記係從表3所選出。 The method of claim 1, wherein the label is selected from Table 3. 如申請專利範圍第4項的方法，其中在該調控基因中的表現至少有2倍差異。 The method of claim 4, wherein the expression in the regulatory gene is at least 2-fold different. 如申請專利範圍第4項的方法，其中表示不同調控的p-值低於0.05。 The method of claim 4, wherein the p-value of the different regulation is less than 0.05. 一種診斷套件，包含適於單離CEC的試劑和適於檢測表2之標記的表現的試劑。 A diagnostic kit comprising reagents suitable for single CEC separation and reagents suitable for detecting the performance of the markers of Table 2. 如申請專利範圍第13項的診斷套件，其中適於檢測標記的該試劑如表3所列者。 A diagnostic kit according to claim 13 wherein the reagent suitable for detecting the label is as listed in Table 3. 如申請專利範圍第13項的套件，進一步包含適於進行一微陣列分析的試劑。 The kit of claim 13 further comprises reagents suitable for performing a microarray analysis. 如申請專利範圍第13項的套件，進一步包含適於放大該標記的試劑。 The kit of claim 13 further comprising a reagent suitable for amplifying the label. 如申請專利範圍第13項的套件，進一步包含含有可執行指南的一成分，其中該指南利用一模式與表現檢測關聯。 The kit of claim 13 further includes a component containing an executable guide, wherein the guide utilizes a pattern to correlate with performance detection. 如申請專利範圍第13項的套件，其中適於單離CEC的該試劑包括免疫磁性試劑。 A kit according to claim 13 wherein the reagent suitable for isolated CEC comprises an immunomagnetic reagent. 一種評估醫療狀況的方法，包含單離CEC、放大從該CEC取得的基因表現標記、檢測該標記的該表現，和利用一預測或一預後與該標記的該表現關聯；其中單離和放大係使用如申請專利範圍第13項的該套件執行。 A method of assessing a medical condition, comprising arranging a single CEC, amplifying a gene expression marker obtained from the CEC, detecting the performance of the marker, and correlating the performance with the marker using a prediction or a prognosis; wherein the singularization and amplification system It is executed using the kit as in claim 13 of the patent application. 一種評估醫療狀況的方法，包含單離CEC、放大從該CEC取得的基因表現標記、檢測該標記的該表現，和利用一預測或一預後與該標記的該表現關聯；其中單離和放大係使用如申請專利範圍第14項的該診斷套件執行。 A method of assessing a medical condition, comprising arranging a single CEC, amplifying a gene expression marker obtained from the CEC, detecting the performance of the marker, and correlating the performance with the marker using a prediction or a prognosis; wherein the singularization and amplification system This diagnostic kit is used as described in claim 14 of the scope of the patent application.
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