KR102618065B1 - Composition for Diagnosing Pancreatic Cancer and Method for Diagnosing Pancreatic Cancer Using the same - Google Patents
Composition for Diagnosing Pancreatic Cancer and Method for Diagnosing Pancreatic Cancer Using the same Download PDFInfo
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- KR102618065B1 KR102618065B1 KR1020210177105A KR20210177105A KR102618065B1 KR 102618065 B1 KR102618065 B1 KR 102618065B1 KR 1020210177105 A KR1020210177105 A KR 1020210177105A KR 20210177105 A KR20210177105 A KR 20210177105A KR 102618065 B1 KR102618065 B1 KR 102618065B1
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- pancreatic cancer
- scg5
- protein
- expression level
- present
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Classifications
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- G—PHYSICS
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
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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
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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
- C12Q2600/00—Oligonucleotides characterized by their use
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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
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Abstract
본 발명은 혈액 내 SCG5 발현 수준 검출을 통한 췌장암 진단용 조성물에 관한 것이다. 본 발명의 췌장암 진단용 조성물 또는 진단 키트를 활용하면 혈액 검사를 통해 매우 신속하고 간편하게 췌장암을 조기 진단할 수 있다. The present invention relates to a composition for diagnosing pancreatic cancer through detection of SCG5 expression level in blood. Using the composition or diagnostic kit for diagnosing pancreatic cancer of the present invention, pancreatic cancer can be diagnosed very quickly and easily through a blood test.
Description
본 발명은 혈액 내 SCG5 발현 수준 검출을 통한 췌장암 진단용 조성물에 관한 것이다. The present invention relates to a composition for diagnosing pancreatic cancer through detection of SCG5 expression level in blood.
췌장암(pancreatic cancer)이란 췌장 세포에서 기원하는 암을 의미하고, 최근 들어 그 발병률이 꾸준히 증가하고 있으며, 특히 한국에서 암 발생 순위 8위(국가암정보센터, 2018년 현황), 사망률 5위(국가암정보센터, 2019년 현황)를 차지하고 있다. 췌장암에는 여러 가지 종류가 있는데 췌관 세포에서 발생한 췌관 선암종이 90% 정도를 차지하고 있어 일반적으로 췌장암이라고 하면 췌관 선암종을 의미한다. 그 외에 낭종성암(낭선암), 내분비종양 등이 있다. 췌장암 환자 중 약 5~10%는 유전 소인을 가지고 있는데, 췌장암 환자에서 췌장암의 가족력이 있는 경우는 약 7.8% 정도로 일반인에서의 췌장암 발생률 0.6%에 비해 빈도가 높다. 췌장암은 5년 생존율이 5% 이하로 예후가 매우 나쁜 암이다. 그 이유는 대부분 암이 진행된 후에 발견되기 때문에 발견 당시 수술 절제가 가능한 경우가 20% 이내이고, 육안으로 보기에 완전히 절제되었다 하더라도 미세 전이에 의해 생존율 향상이 적으며, 항암제 및 방사선 치료에 대한 반응이 낮기 때문이다. 따라서, 생존율을 향상시킬 수 있는 가장 중요한 방법은 증상이 없거나 비특이적일 때 조기 발견하여 수술하는 것이다. Pancreatic cancer refers to cancer that originates from pancreatic cells, and its incidence rate has been steadily increasing in recent years. In particular, in Korea, it ranks 8th in cancer incidence (National Cancer Information Center, 2018 status) and 5th in mortality (national cancer information center). Cancer Information Center, 2019 status). There are several types of pancreatic cancer, and pancreatic adenocarcinoma, which arises from pancreatic duct cells, accounts for about 90%, so pancreatic cancer generally refers to pancreatic ductal adenocarcinoma. In addition, there are cystic cancer (cystadenocarcinoma) and endocrine tumors. Approximately 5-10% of pancreatic cancer patients have a genetic predisposition, and approximately 7.8% of pancreatic cancer patients have a family history of pancreatic cancer, which is higher than the 0.6% incidence of pancreatic cancer in the general population. Pancreatic cancer is a cancer with a very poor prognosis, with a 5-year survival rate of less than 5%. This is because most cancers are discovered after they have progressed, so surgical resection is possible in less than 20% of cases at the time of discovery, and even if the cancer is completely resected with the naked eye, the improvement in survival rate is small due to micrometastases, and the response to anticancer drugs and radiation therapy is low. Because it is low. Therefore, the most important way to improve survival rate is to detect early and perform surgery when there are no symptoms or when symptoms are non-specific.
이러한 췌장암의 진단을 위해 사용하는 검사에는 복부 초음파, 복부 컴퓨터 단층촬영(CT), 자기공명영상(MRI), 내시경역행담췌관조영술(ERCP), 내시경 초음파(EUS), 양전자 방출 단층촬영(PET scan), 혈청 종양 표지자(CA19-9), 조직검사 등이 있는데, 이들은 비용이 많이 들고 통증을 수반하며 병원에 입원해서 시행해야 하고, 또 임상에서는 실제 증상이 있거나 췌장암 진단이 가능한 환자들에게 실시하는 게 보통이다. Tests used to diagnose pancreatic cancer include abdominal ultrasound, abdominal computed tomography (CT), magnetic resonance imaging (MRI), endoscopic retrograde cholangiopancreatography (ERCP), endoscopic ultrasound (EUS), and positron emission tomography (PET scan). ), serum tumor marker (CA19-9), and biopsy, but these are expensive, painful, and must be performed by hospitalization in a hospital. In addition, in clinical practice, they are performed on patients who have actual symptoms or can be diagnosed with pancreatic cancer. It's normal.
기존 췌장암 진단에 이용하고 있는 탄수화물 항원(carbohydrate antigen) 19-9(CA19-9) 검사법은 암세포에 의해 분비되는 CA19-9의 양을 측정해 암의 경과를 추적하는 방법으로서 70 내지 80%의 민감도와 80 내지 90%의 특이도를 나타내고 진단 정확도(AUC)는 77%이다. The carbohydrate antigen 19-9 (CA19-9) test, which is currently used to diagnose pancreatic cancer, is a method of tracking the progress of cancer by measuring the amount of CA19-9 secreted by cancer cells and has a sensitivity of 70 to 80%. It has a specificity of 80 to 90% and a diagnostic accuracy (AUC) of 77%.
다만, CA19-9법은 췌장암뿐만 아니라 담도를 포함한 소화기계 암에서 모두 상승할 수 있고 담관염과 담도 폐색이 있는 경우에도 상승할 수 있다. 또한, 결장직장암, 폐암, 간질환 등과 같은 다른 암과 다른 질환에서도 증가할 수 있다. 특히 조기암에서는 정상 수치를 나타내는 경우가 많아 조기 진단에 사용하기 어려운 문제점이 있다. However, CA19-9 can be elevated not only in pancreatic cancer but also in cancers of the digestive system including the biliary tract, and can also be elevated in cases of cholangitis and biliary obstruction. It may also increase in other cancers and other diseases, such as colorectal cancer, lung cancer, and liver disease. In particular, early cancer often shows normal values, making it difficult to use for early diagnosis.
본 발명자들은 증상이 없는 췌장암 초기 환자들이 혈액검사에 의한 선별검사를 통해 췌장암을 조기에 진단할 수 있는 진단 방법을 규명하기 위해 예의 연구 노력하였다. 그 결과 혈액 내 SCG5 수준 검출을 통해 췌장암의 조기 진단이 가능함을 규명함으로써, 본 발명을 완성하게 되었다. The present inventors have made extensive research efforts to identify a diagnostic method by which early stage pancreatic cancer patients without symptoms can be diagnosed with pancreatic cancer through blood test screening. As a result, the present invention was completed by demonstrating that early diagnosis of pancreatic cancer is possible through detection of SCG5 levels in the blood.
따라서, 본 발명의 목적은 췌장암 진단용 조성물을 제공하는 것이다. Therefore, an object of the present invention is to provide a composition for diagnosing pancreatic cancer.
본 발명의 다른 목적은 췌장암 진단 키트를 제공하는 것이다.Another object of the present invention is to provide a pancreatic cancer diagnostic kit.
본 발명의 다른 목적은 췌장암 진단 방법을 제공하는 것이다. Another object of the present invention is to provide a method for diagnosing pancreatic cancer.
본 발명의 다른 목적은 췌장암 치료제 스크리닝 방법을 제공하는 것이다.Another object of the present invention is to provide a screening method for pancreatic cancer treatment.
본 발명의 일 양태에 따르면, 본 발명은 SCG5 단백질 또는 상기 SCG5 단백질을 암호화하는 유전자의 mRNA 발현 수준을 검출하는 제제를 포함하는 췌장암 진단용 조성물을 제공한다.According to one aspect of the present invention, the present invention provides a composition for diagnosing pancreatic cancer comprising an agent for detecting the mRNA expression level of SCG5 protein or the gene encoding the SCG5 protein.
본 발명자들은 증상이 없는 췌장암 초기 환자들이 혈액검사에 의한 선별검사를 통해 췌장암을 조기에 진단할 수 있는 진단 방법을 규명하기 위해 예의 연구 노력하였다. 그 결과 혈액 내 SCG5 수준 검출을 통해 췌장암의 조기 진단이 가능함을 규명하였다. The present inventors have made extensive research efforts to identify a diagnostic method by which early stage pancreatic cancer patients without symptoms can be diagnosed with pancreatic cancer through blood test screening. As a result, it was found that early diagnosis of pancreatic cancer is possible through detection of SCG5 levels in the blood.
본 발명의 조성물을 이용하면, 정상군과 췌장암의 환자군을 구별하여 검출 또는 진단할 수 있고, 나아가, 다른 종류의 암과 구별하여 췌장암을 선별적으로 검출 또는 진단할 수 있다. 본 발명에서 용어 "진단"은 특정 질병 또는 질환에 대한 대상(subject)의 감수성(susceptibility)을 판정하는 것 또는 대상이 특정 질병 또는 질환을 현재 가지고 있는지 여부를 판정하는 것을 의미한다. Using the composition of the present invention, pancreatic cancer can be detected or diagnosed by distinguishing between a normal group and a pancreatic cancer patient group, and furthermore, pancreatic cancer can be selectively detected or diagnosed by distinguishing it from other types of cancer. In the present invention, the term “diagnosis” means determining a subject's susceptibility to a specific disease or condition, or determining whether the subject currently has a specific disease or condition.
본 발명의 췌장암 진단용 조성물은 SCG5 단백질 또는 상기 SCG5 단백질을 암호화하는 유전자의 mRNA의 발현 수준을 측정하는 제제를 포함할 수 있다.The composition for diagnosing pancreatic cancer of the present invention may include an agent for measuring the expression level of SCG5 protein or mRNA of the gene encoding the SCG5 protein.
본 발명의 SCG5 단백질 또는 이를 암호화하는 유전자의 mRNA는 대상(subject)으로부터 분리된 생물학적 시료, 보다 구체적으로 예를 들면, 혈액, 혈청 또는 혈장에 존재하는 것일 수 있다.The SCG5 protein of the present invention or the mRNA of the gene encoding it may be present in a biological sample isolated from a subject, more specifically, for example, blood, serum or plasma.
본 발명의 일 구현예에 있어서, 본 발명에서의 SCG5 단백질 수준은 인체로부터 분리된 혈액 시료 내의 SCG5 단백질 수준을 의미한다.In one embodiment of the present invention, the SCG5 protein level in the present invention refers to the SCG5 protein level in a blood sample isolated from the human body.
종래 알려진 췌장암의 진단을 위해 사용하는 검사로서, 복부 초음파, 복부 컴퓨터 단층촬영(CT), 자기공명영상(MRI), 내시경역행담췌관조영술(ERCP), 내시경 초음파(EUS), 양전자 방출 단층촬영(PET scan), 혈청 종양 표지자(CA19-9), 조직검사 등을 예로 들 수 있고, 이들은 비용이 많이 들고 통증을 수반하며 병원에 입원해서 시행해야 하고, 또 임상에서는 실제 증상이 있거나 췌장암 진단이 가능한 환자들에게 실시하는 게 보통인 한계가 있었다. Conventionally known tests used to diagnose pancreatic cancer include abdominal ultrasound, abdominal computed tomography (CT), magnetic resonance imaging (MRI), endoscopic retrograde cholangiopancreatography (ERCP), endoscopic ultrasound (EUS), and positron emission tomography ( Examples include PET scan, serum tumor marker (CA19-9), and biopsy. These are expensive, painful, and must be performed by hospitalization at a hospital. In addition, in clinical practice, pancreatic cancer can be diagnosed when there are actual symptoms or when it is possible to diagnose pancreatic cancer. There were limitations to how it was usually performed on patients.
이에 반해 본 발명의 일 실시예에 따른 췌장암 진단용 조성물을 이용하는 경우, 대상으로부터 분리된 혈액, 혈청 또는 혈장에 있어서, SCG5 단백질 또는 상기 단백질을 암호화하는 유전자의 mRNA의 발현 수준을 측정함으로써 신속하고 간편하면서도 정확하게 췌장암을 조기 진단할 수 있다는 장점이 있다.On the other hand, when using the composition for diagnosing pancreatic cancer according to an embodiment of the present invention, the expression level of the SCG5 protein or the mRNA of the gene encoding the protein is measured in blood, serum, or plasma isolated from the subject, which is quick and convenient. It has the advantage of accurately diagnosing pancreatic cancer at an early stage.
본 발명의 일 구현예에 있어서, 본 발명의 SCG5 단백질 수준을 검출하는 제제는 SCG5 단백질에 특이적으로 결합하는 항체, 앱타머, 리간드, PNA(peptide nucleic acid) 및 올리고펩타이드로 구성된 군으로부터 선택되는 어느 하나 이상이다. In one embodiment of the present invention, the agent for detecting SCG5 protein level of the present invention is selected from the group consisting of antibodies, aptamers, ligands, PNA (peptide nucleic acid), and oligopeptides that specifically bind to SCG5 protein. It's more than one thing.
본 발명의 "단백질 수준을 검출"이란 췌장암을 진단하기 위하여 생물학적 시료인 혈액에서 본 발명에서의 췌장암 진단용 마커인 SCG5 단백질의 존재 여부와 발현 정도를 확인하는 과정을 의미한다. 상기 단백질의 발현 수준 측정 또는 비교 분석 방법으로는 단백질 칩 분석, 면역측정법, 리간드 바인딩 어세이, MALDI-TOF(Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) 분석, SELDI-TOF(Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) 분석, 방사선 면역분석, 방사 면역 확산법, 오우크테로니 면역 확산법, 로케트 면역전기영동, 조직면역 염색, 보체 고정 분석법, 2차원 전기영동 분석, 액상 크로마토그래피-질량분석(liquid chromatography-Mass Spectrometry, LC-MS), LC-MS/MS(liquid chromatography-Mass Spectrometry/ Mass Spectrometry), 웨스턴 블랏팅 및 ELISA(enzyme linked immunosorbentassay) 등이 있으나, 이에 제한되는 것은 아니다.“Detecting the protein level” of the present invention refers to the process of confirming the presence and expression level of SCG5 protein, a marker for pancreatic cancer diagnosis in the present invention, in blood, a biological sample, in order to diagnose pancreatic cancer. Methods for measuring or comparatively analyzing the expression level of the protein include protein chip analysis, immunoassay, ligand binding assay, Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF) analysis, and Surface Enhanced Laser Desorption (SELDI-TOF). /Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion method, Ouchteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoresis analysis, liquid chromatography-mass spectrometry. (liquid chromatography-Mass Spectrometry, LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/Mass Spectrometry), Western blotting, and ELISA (enzyme linked immunosorbent assay), etc., but are not limited thereto.
본 발명의 "항체"는 항원과 특이적으로 결합하여 항원-항체 반응을 일으키는 물질을 의미한다. 본 발명의 목적상, 항체는 SCG5 단백질에 대해 특이적으로 결합하는 항체를 의미한다. 본 발명의 항체는 다클론 항체, 단클론 항체 및 재조합 항체를 모두 포함한다. 상기 항체는 당업계에 널리 공지된 기술을 이용하여 용이하게 제조될 수 있다. 예를 들어, 다클론 항체는 상기 SCG5 단백질 항원을 동물에 주사하고 동물로부터 채혈하여 항체를 포함하는 혈청을 수득하는 과정을 포함하는 당업계에 널리 공지된 방법에 의해 생산될 수 있다. 이러한 다클론 항체는 염소, 토끼, 양, 원숭이, 말, 돼지, 소, 개 등의 임의의 동물로부터 제조될 수 있다. 또한, 단클론 항체는 당업계에 널리 공지된 하이브리도마 방법(hybridoma method; Kohler 및 Milstein (1976) European Journal of Immunology 6:511-519 참조), 또는 파지 항체 라이브러리 기술(Clackson et al, Nature, 352:624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991 참조)을 이용하여 제조될 수 있다. 상기 방법으로 제조된 항체는 겔 전기영동, 투석, 염 침전, 이온교환 크로마토그래피, 친화성 크로마토그래피 등의 방법을 이용하여 분리, 정제될 수 있다. 또한, 본 발명의 항체는 2개의 전장의 경쇄 및 2개의 전장의 중쇄를 갖는 완전한 형태뿐만 아니라, 항체 분자의 기능적인 단편을 포함한다. 항체 분자의 기능적인 단편이란, 적어도 항원 결합 기능을 보유하고 있는 단편을 의미하며, Fab, F(ab'), F(ab')2 및 Fv 등이 있다.“Antibody” of the present invention refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, antibody refers to an antibody that specifically binds to the SCG5 protein. Antibodies of the present invention include polyclonal antibodies, monoclonal antibodies, and recombinant antibodies. The antibody can be easily produced using techniques well known in the art. For example, polyclonal antibodies can be produced by a method well known in the art, which includes injecting the SCG5 protein antigen into an animal and collecting blood from the animal to obtain serum containing the antibody. These polyclonal antibodies can be produced from any animal, such as goats, rabbits, sheep, monkeys, horses, pigs, cows, dogs, etc. In addition, monoclonal antibodies can be prepared using the hybridoma method (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), which is well known in the art, or phage antibody library technology (Clackson et al, Nature, 352 :624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). Antibodies prepared by the above method can be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. Additionally, antibodies of the invention include intact forms with two full-length light chains and two full-length heavy chains, as well as functional fragments of the antibody molecule. A functional fragment of an antibody molecule refers to a fragment that possesses at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2, and Fv.
본 발명의 용어 "앱타머"는 올리고핵산 또는 펩타이드 분자이며, 앱타머의 일반적인 내용은 문헌[Bock LC et al., Nature 355(6360):5646(1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630(2000); Cohen BA, Colas P, Brent R. "An artificial cell-cycle inhibitor isolated from a combinatorial library". Proc Natl Acad Sci USA. 95(24): 142727(1998)]에 상세하게 개시되어 있다.As used herein, the term “aptamer” refers to an oligonucleic acid or peptide molecule, and general information on aptamers is described in Bock LC et al., Nature 355(6360):5646 (1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630(2000); Cohen BA, Colas P, Brent R. “An artificial cell-cycle inhibitor isolated from a combinatorial library”. Proc Natl Acad Sci USA. 95(24): 142727 (1998)].
본 발명의 용어 “리간드”는 SCG5 단백질에 결합하여 복합체를 형성하고 신호를 발생시킬 수 있는 저분자 물질을 의미한다. 포괄적으로 항체를 포함하는 의미로 해석될 수도 있다. The term “ligand” of the present invention refers to a small molecule that can bind to the SCG5 protein to form a complex and generate a signal. It may be interpreted to comprehensively include antibodies.
본 발명의 용어 "PNA(Peptide Nucleic Acid)"는 인공적으로 합성된, DNA 또는 RNA와 비슷한 중합체를 가리키며, 1991년 덴마크 코펜하겐 대학교의 Nielsen, Egholm, Berg와 Buchardt 교수에 의해 처음으로 소개되었다. DNA는 인산-리보스당 골격을 갖는데 반해, PNA는 펩타이드 결합에 의해 연결된 반복된 N-(2-아미노에틸)-글리신 골격을 가지며, 이로 인해 DNA 또는 RNA에 대한 결합력과 안정성이 크게 증가되어 분자 생물학, 진단 분석 및 안티센스 치료법에 사용되고 있다. PNA는 문헌 [P E Nielsen, M Egholm, R H Berg, O Buchardt, "Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide", Science 254(5037):1497-500 (1991)]에 상세하게 개시되어 있다.The term "Peptide Nucleic Acid (PNA)" of the present invention refers to an artificially synthesized polymer similar to DNA or RNA, and was first introduced by Professors Nielsen, Egholm, Berg and Buchardt at the University of Copenhagen, Denmark in 1991. While DNA has a phosphate-ribose sugar backbone, PNA has a repeated N-(2-aminoethyl)-glycine backbone linked by peptide bonds, which greatly increases its binding force and stability to DNA or RNA, making it useful in molecular biology. , is used in diagnostic analysis and antisense therapy. PNA is described in detail in P E Nielsen, M Egholm, R H Berg, O Buchardt, "Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide", Science 254(5037):1497-500 (1991). It has been disclosed.
본 발명의 일 실시예에 있어서, 본 발명의 SCG5 단백질을 암호화하는 유전자의 mRNA 발현 수준을 검출하는 제제는 상기 mRNA에 특이적으로 결합하는 프라이머, 프로브 및 안티센스 뉴클레오티드로 이루어진 군으로부터 선택되는 어느 하나 이상이다. 본 발명에 따른 SCG5 단백질의 정보는 알려져 있으므로, 당업자라면 이를 바탕으로 상기 단백질을 암호화하는 유전자의 mRNA에 특이적으로 결합하는 프라이머, 프로브 또는 안티센스 뉴클레오티드를 용이하게 디자인할 수 있을 것이다.In one embodiment of the present invention, the agent for detecting the mRNA expression level of the gene encoding the SCG5 protein of the present invention contains at least one selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the mRNA. am. Since information on the SCG5 protein according to the present invention is known, a person skilled in the art will be able to easily design primers, probes, or antisense nucleotides that specifically bind to the mRNA of the gene encoding the protein based on this information.
본 발명의 용어 "mRNA 발현 수준을 검출"이란 췌장암을 진단하기 위하여 생물학적 시료에서 SCG5 단백질을 암호화하는 유전자들의 mRNA 존재 여부와 발현 정도를 확인하는 과정으로 mRNA의 양을 측정하는 것을 의미한다. 이를 위한 분석 방법으로는 역전사 중합효소반응(RT-PCR), 경쟁적 역전사 중합효소반응(Competitive RT-PCR), 실시간 역전사 중합효소반응(Real-time RT-PCR), RNase 보호 분석법(RPA; RNase protection assay), 노던 블랏팅(Northern blotting), DNA 칩 등이 있으나 이에 제한되는 것은 아니다.The term "detection of mRNA expression level" in the present invention refers to measuring the amount of mRNA through the process of confirming the presence and expression level of mRNA of genes encoding the SCG5 protein in a biological sample in order to diagnose pancreatic cancer. Analysis methods for this include reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction (Competitive RT-PCR), real-time reverse transcription polymerase reaction (Real-time RT-PCR), and RNase protection assay (RPA). assay), Northern blotting, DNA chip, etc., but are not limited to these.
본 발명의 용어 "프라이머"는 표적 유전자 서열을 인지하는 단편으로서, 정방향 및 역방향의 프라이머 쌍을 포함하나, 바람직하게는, 특이성 및 민감성을 가지는 분석 결과를 제공하는 프라이머 쌍이다. 프라이머의 핵산 서열이 시료 내 존재하는 비-표적 서열과 불일치하는 서열이어서, 상보적인 프라이머 결합 부위를 함유하는 표적 유전자 서열만 증폭하고 비특이적 증폭을 유발하지 않는 프라이머일 때, 높은 특이성이 부여될 수 있다.The term “primer” in the present invention refers to a fragment that recognizes a target gene sequence and includes forward and reverse primer pairs, but is preferably a primer pair that provides analysis results with specificity and sensitivity. High specificity can be granted when the nucleic acid sequence of the primer is a sequence that is inconsistent with the non-target sequence present in the sample, so that the primer amplifies only the target gene sequence containing the complementary primer binding site and does not cause non-specific amplification. .
본 발명의 용어 "프로브"란 시료 내의 검출하고자 하는 표적 물질과 특이적으로 결합할 수 있는 물질을 의미하며, 상기 결합을 통하여 특이적으로 시료 내의 표적 물질의 존재를 확인할 수 있는 물질을 의미한다. 프로브의 종류는 당업계에서 통상적으로 사용되는 물질이면 특별히 제한되지 않으나, 바람직하게는 PNA(peptide nucleic acid), LNA(locked nucleic acid), 펩타이드, 폴리펩타이드, 단백질, RNA 또는 DNA일 수 있으며, 가장 바람직하게는 PNA이다. 보다 구체적으로, 상기 프로브는 바이오 물질로서 생물에서 유래되거나 이와 유사한 것 또는 생체 외에서 제조된 것을 포함하는 것으로, 예를 들어, 효소, 단백질, 항체, 미생물, 동식물 세포 및 기관, 신경세포, DNA 및 RNA일 수 있으며, DNA는 cDNA, 게놈 DNA, 올리고뉴클레오타이드를 포함하며, RNA는 게놈 RNA, mRNA, 올리고뉴클레오타이드를 포함하며, 단백질의 예로는 항체, 항원, 효소, 펩타이드 등을 포함할 수 있다.The term "probe" in the present invention refers to a substance that can specifically bind to a target substance to be detected in a sample, and refers to a substance that can specifically confirm the presence of the target substance in the sample through said binding. The type of probe is not particularly limited as long as it is a material commonly used in the art, but is preferably PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA, or DNA. Preferably it is PNA. More specifically, the probe is a biomaterial that is derived from or similar to living organisms or includes those manufactured in vitro, such as enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, nerve cells, DNA and RNA. DNA may include cDNA, genomic DNA, and oligonucleotides, RNA may include genomic RNA, mRNA, and oligonucleotides, and examples of proteins may include antibodies, antigens, enzymes, peptides, etc.
본 발명의 용어 "LNA(Locked nucleic acids)"는, 2'-O, 4'-C 메틸렌 브릿지를 포함하는 핵산 아날로그를 의미한다 [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502]. LNA 뉴클레오사이드는 DNA와 RNA의 일반적 핵산 염기를 포함하며, 왓슨-크릭 염기 쌍 규칙에 따라 염기 쌍을 형성할 수 있다. 하지만, 메틸렌 브릿지로 인한 분자의 'locking'으로 인해, LNA는 왓슨-크릭 결합에서 이상적 형상을 형성하지 못하게 된다. LNA가 DNA 또는 RNA 올리고뉴클레오티드에 포함되면, LNA는 보다 빠르게 상보적 뉴클레오티드 사슬과 쌍을 이루어 이중 나선의 안정성을 높일 수 있다. The term "LNA (Locked nucleic acids)" of the present invention refers to a nucleic acid analog containing 2'-O, 4'-C methylene bridges [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502 ]. LNA nucleosides contain the common nucleic acid bases of DNA and RNA and can form base pairs according to the Watson-Crick base pairing rules. However, due to the 'locking' of the molecule due to the methylene bridge, LNA does not form the ideal shape in the Watson-Crick bond. When LNA is included in a DNA or RNA oligonucleotide, the LNA can pair with the complementary nucleotide chain more quickly and increase the stability of the double helix.
본 발명의 용어 "안티센스"는 안티센스 올리고머가 왓슨-크릭 염기쌍 형성에 의해 RNA 내의 표적 서열과 혼성화되어, 표적서열 내에서 전형적으로 mRNA와 RNA:올리고머 헤테로이중체의 형성을 허용하는, 뉴클레오티드 염기의 서열 및 서브유닛간 백본을 갖는 올리고머를 의미한다. 올리고머는 표적 서열에 대한 정확한 서열 상보성 또는 근사 상보성을 가질 수 있다. The term "antisense" of the present invention refers to a sequence of nucleotide bases in which an antisense oligomer hybridizes with a target sequence in RNA by Watson-Crick base pairing, typically allowing the formation of an mRNA and RNA:oligomer heteroduplex within the target sequence. and an oligomer having an intersubunit backbone. Oligomers may have exact or approximate sequence complementarity to the target sequence.
본 발명의 다른 일 양태에 따르면, 본 발명은 상술한 본 발명의 다른 일 양태에 따른 췌장암 진단용 조성물을 포함하는 췌장암 진단용 키트를 제공한다. According to another aspect of the present invention, the present invention provides a kit for diagnosing pancreatic cancer, including the composition for diagnosing pancreatic cancer according to another aspect of the present invention described above.
본 발명의 용어 “키트”는 RT-PCR 키트, DNA 칩 키트, ELISA 키트, 단백질 칩 키트, 래피드(rapid) 키트 또는 MRM(Multiple reaction monitoring) 키트일 수 있으나, 이에 제한되는 것은 아니다.The term “kit” in the present invention may be, but is not limited to, an RT-PCR kit, DNA chip kit, ELISA kit, protein chip kit, rapid kit, or MRM (multiple reaction monitoring) kit.
본 발명의 췌장암 진단용 키트는 분석 방법에 적합한 한 종류 또는 그 이상의 다른 구성성분 조성물, 용액 또는 장치를 더 포함할 수 있다.The kit for diagnosing pancreatic cancer of the present invention may further include one or more other component compositions, solutions, or devices suitable for the analysis method.
예를 들면, 상기 진단용 키트는 역전사 중합효소반응을 수행하기 위해 필요한 필수 요소를 더 포함할 수 있다. 역전사 중합효소반응 키트는 마커 단백질을 암호화하는 유전자에 대해 특이적인 프라이머 쌍을 포함한다. 프라이머는 상기 유전자의 핵산서열에 특이적인 서열을 가지는 뉴클레오타이드로서, 약 7 bp 내지 50 bp의 길이, 보다 바람직하게는 약 10 bp 내지 30 bp의 길이를 가질 수 있다. 또한 대조군 유전자의 핵산 서열에 특이적인 프라이머를 포함할 수 있다. 그 외 역전사 중합효소반응 키트는 테스트 튜브 또는 다른 적절한 용기, 반응 완충액(pH 및 마그네슘 농도는 다양), 데옥시뉴클레오타이드(dNTPs), Taq-폴리머라아제 및 역전사효소와 같은 효소, DNase, RNase 억제제 DEPC-수(DEPC-water), 멸균수 등을 포함할 수 있다.For example, the diagnostic kit may further include essential elements required to perform a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit contains a pair of primers specific for the gene encoding the marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene, and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. It may also include primers specific to the nucleic acid sequence of the control gene. Other reverse transcription polymerase reaction kits include test tubes or other suitable containers, reaction buffer (pH and magnesium concentration vary), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, and the RNase inhibitor DEPC. -Can include DEPC-water, sterilized water, etc.
또한, 본 발명의 진단용 키트는 DNA 칩을 수행하기 위해 필요한 필수 요소를 포함할 수 있다. DNA 칩 키트는 유전자 또는 그의 단편에 해당하는 cDNA 또는 올리고뉴클레오티드(oligonucleotide)가 부착되어 있는 기판, 및 형광표지 프로브를 제작하기 위한 시약, 제제, 효소 등을 포함할 수 있다. 또한 기판은 대조군 유전자 또는 그의 단편에 해당하는 cDNA 또는 올리고뉴클레오티드를 포함할 수 있다.Additionally, the diagnostic kit of the present invention may include essential elements required to perform DNA chip testing. A DNA chip kit may include a substrate to which a cDNA or oligonucleotide corresponding to a gene or a fragment thereof is attached, and reagents, agents, enzymes, etc. for producing a fluorescent label probe. The substrate may also include cDNA or oligonucleotides corresponding to control genes or fragments thereof.
또한, 본 발명의 진단용 키트는 ELISA를 수행하기 위해 필요한 필수 요소를 포함할 수 있다. ELISA 키트는 상기 단백질에 대해 특이적인 항체를 포함한다. 항체는 마커 단백질에 대한 특이성 및 친화성이 높고 다른 단백질에 대한 교차 반응성이 거의 없는 항체로, 단클론 항체, 다클론 항체 또는 재조합 항체이다. 또한 ELISA 키트는 대조군 단백질에 특이적인 항체를 포함할 수 있다. 그 외 ELISA 키트는 결합된 항체를 검출할 수 있는 시약, 예를 들면, 표지된 2차 항체, 발색단(chromophores), 효소(예: 항체와 컨주게이트됨) 및 그의 기질 또는 항체와 결합할 수 있는 다른 물질 등을 포함할 수 있다.Additionally, the diagnostic kit of the present invention may include essential elements required to perform ELISA. ELISA kits contain antibodies specific for these proteins. Antibodies are antibodies that have high specificity and affinity for a marker protein and almost no cross-reactivity to other proteins, and may be monoclonal antibodies, polyclonal antibodies, or recombinant antibodies. Additionally, ELISA kits may include antibodies specific for control proteins. Other ELISA kits include reagents that can detect bound antibodies, such as labeled secondary antibodies, chromophores, enzymes (e.g., conjugated with antibodies) and their substrates or those that can bind to antibodies. It may contain other substances, etc.
본 발명의 “췌장암 진단용 키트”는 상술한 본 발명의 다른 일 양태인 “췌장암 진단용 조성물”을 포함하는 바, 중복되는 내용을 원용하며, 본 명세서 기재의 과도한 복잡성을 피하기 위해 중복 기재를 생략하도록 한다. The “kit for diagnosing pancreatic cancer” of the present invention includes the “composition for diagnosing pancreatic cancer,” which is another aspect of the present invention described above. Redundant content is used, and redundant description is omitted to avoid excessive complexity of the description in this specification. .
본 발명의 다른 일 양태에 따르면, 본 발명은 다음 단계를 포함하는 췌장암 진단 방법을 제공한다:According to another aspect of the present invention, the present invention provides a method for diagnosing pancreatic cancer comprising the following steps:
(a) 인체로부터 분리된 혈액 시료 내의 SCG5 단백질 수준 또는 SCG5 단백질을 암호화하는 유전자의 mRNA 발현 수준을 측정하는 단계; 정상 혈액 대조군 시료에 비해 상기 SCG5 단백질 수준 또는 상기 mRNA 발현 수준이 감소된 경우, 췌장암으로 진단함.(a) measuring the SCG5 protein level or the mRNA expression level of the gene encoding the SCG5 protein in a blood sample isolated from the human body; When the SCG5 protein level or the mRNA expression level is decreased compared to a normal blood control sample, pancreatic cancer is diagnosed.
본 발명의 일 구현예에 있어서, 본 발명의 SCG5 단백질 발현 수준을 측정하는 단계는 SCG5 단백질에 특이적으로 결합하는 항체, 앱타머, 리간드, PNA(peptide nucleic acid) 및 올리고펩타이드로 구성된 군으로부터 선택되는 어느 하나 이상인 SCG5 단백질 수준을 검출하는 제제를 이용하여 수행된다.In one embodiment of the present invention, the step of measuring the SCG5 protein expression level of the present invention is selected from the group consisting of antibodies, aptamers, ligands, PNA (peptide nucleic acids), and oligopeptides that specifically bind to the SCG5 protein. This is performed using an agent that detects one or more SCG5 protein levels.
구체적으로 예를 들면, 본 발명의 SCG5 단백질에 특이적으로 결합하는 항체를 이용하여 진단 대상 혈액 시료 및 정상 혈액 대조군 시료의 SCG5 단백질 발현을 측정 및 비교할 수 있다. 상기 항체와 생물학적 시료 내의 SCG5 단백질이 항원-항체 복합체를 형성하도록 하고, 이를 검출하는 방법을 이용한다. Specifically, for example, the SCG5 protein expression of blood samples to be diagnosed and normal blood control samples can be measured and compared using the antibody that specifically binds to the SCG5 protein of the present invention. The antibody and the SCG5 protein in the biological sample form an antigen-antibody complex, and a method is used to detect this.
본 발명의 용어 "항원-항체 복합체"는 생물학적 시료 중의 해당 유전자의 존재 또는 부존재를 확인하기 위한 단백질 항원과 이를 인지하는 항체의 결합물을 의미한다. 상기 항원-항체 복합체의 검출은 당업계에 공지된 방법, 예를 들어 분광학적, 광화학적, 생물화학적, 면역화학적, 전기적, 흡광적, 화학적 방법 및 기타 다양한 방법을 이용하여 검출할 수 있다.The term "antigen-antibody complex" of the present invention refers to a combination of a protein antigen and an antibody that recognizes it to confirm the presence or absence of the corresponding gene in a biological sample. The antigen-antibody complex can be detected using methods known in the art, such as spectroscopic, photochemical, biochemical, immunochemical, electrical, absorbance, chemical, and other methods.
본 발명의 일 구현예에 있어서, 본 발명의 SCG5 단백질을 암호화하는 유전자의 mRNA 발현 수준을 측정하는 단계는 상기 mRNA에 특이적으로 결합하는 프라이머, 프로브 및 안티센스 뉴클레오티드로 이루어진 군으로부터 선택되는 어느 하나 이상인 mRNA 발현 수준을 검출하는 제제를 이용하여 수행된다.In one embodiment of the present invention, the step of measuring the mRNA expression level of the gene encoding the SCG5 protein of the present invention is at least one selected from the group consisting of a primer, a probe, and an antisense nucleotide that specifically binds to the mRNA. This is performed using an agent that detects mRNA expression levels.
구체적으로 예를 들면, 본 발명의 SCG5 단백질을 암호화하는 유전자의 mRNA의 발현 수준의 측정 또는 비교는 역전사효소 중합효소반응, 경쟁적 역전사효소 중합효소반응, 실시간 역전사효소 중합효소반응, RNase 보호 분석법, 노던 블랏팅 또는 DNA 칩 등을 사용할 수 있으나, 이에 제한되는 것은 아니다. 상기 측정 방법들을 통하여 정상 대조군에서의 mRNA 발현량과 질환 발병 여부를 진단하고자 하는 개체의 mRNA 발현량을 확인할 수 있고, 이들 발현량 정도를 비교함으로써 췌장암의 발병 여부를 진단 또는 예측할 수 있다.Specifically, for example, measurement or comparison of the expression level of mRNA of the gene encoding the SCG5 protein of the present invention can be performed using reverse transcriptase polymerase reaction, competitive reverse transcriptase polymerase reaction, real-time reverse transcriptase polymerase reaction, RNase protection assay, Northern Blotting or DNA chips may be used, but are not limited thereto. Through the above measurement methods, the mRNA expression level in the normal control group and the mRNA expression level in the individual for whom the disease is to be diagnosed can be confirmed, and by comparing these expression levels, the occurrence of pancreatic cancer can be diagnosed or predicted.
본 발명에서 정상 혈액 대조군 시료에 비해, 대상(subject)으로부터 분리한 혈액 시료 내의 SCG5 단백질 수준 또는 상기 mRNA 발현 수준이 감소된 경우, 췌장암으로 진단한다. 보다 구체적으로 예를 들면, 분석 대상 혈액 시료에서의 SCG5 단백질 수준이 106.27 ng/ml 미만으로 측정되거나 또는 정상 혈액 대조군 시료에 비해, mRNA 발현 수준이 20% 이상, 더욱 구체적으로 25% 이상, 더욱 더 구체적으로 30% 이상, 더욱 더 구체적으로 35% 이상, 더욱 더 구체적으로 40% 이상 감소된 경우, 췌장암으로 진단한다. 데이터 특성 상 log2로 변환한 값으로 계산시, 구체적으로 예를 들면, 5% 이상, 더 구체적으로 8% 이상, 더욱 더 구체적으로 9% 이상, 더욱 더 구체적으로 10%이상, 더욱 더 구체적으로 11% 이상 감소된 경우, 췌장암으로 진단할 수 있다. In the present invention, pancreatic cancer is diagnosed when the SCG5 protein level or the mRNA expression level in a blood sample isolated from a subject is decreased compared to a normal blood control sample. More specifically, for example, the SCG5 protein level in the blood sample being analyzed is measured to be less than 106.27 ng/ml, or the mRNA expression level is 20% or more, more specifically 25% or more, or even more, compared to a normal blood control sample. If it is specifically reduced by more than 30%, more specifically by more than 35%, and even more specifically by more than 40%, it is diagnosed as pancreatic cancer. Due to the nature of the data, when calculating with log2 converted values, specifically, for example, 5% or more, more specifically 8% or more, even more specifically 9% or more, even more specifically 10% or more, even more specifically 11 If it decreases by more than %, it can be diagnosed as pancreatic cancer.
본 발명의 췌장암 진단 방법은 상술한 본 발명의 다른 일 양태인 췌장암 진단용 조성물 또는 췌장암 진단용 키트를 이용하여 실시하는 진단 방법에 관한 것인 바, 중복되는 내용을 원용하며, 본 명세서 기재의 과도한 복잡성을 피하기 위해 중복 기재를 생략하도록 한다.The method for diagnosing pancreatic cancer of the present invention relates to a diagnostic method performed using the composition for diagnosing pancreatic cancer or the kit for diagnosing pancreatic cancer, which is another aspect of the present invention described above. Redundant content is cited and the excessive complexity of the description in the present specification is avoided. To avoid duplicate entries, omit them.
본 발명의 다른 일 양태에 따르면, 본 발명은 다음 단계를 포함하는 췌장암 치료제의 스크리닝 방법을 제공한다:According to another aspect of the present invention, the present invention provides a screening method for a therapeutic agent for pancreatic cancer comprising the following steps:
(a) 혈액 내 SCG5 단백질 또는 SCG5 mRNA 발현 수준이 증가되어 있는 췌장암 동물 질병 모델에 대해 의약 후보 물질을 투여하는 단계; 및(a) administering a drug candidate to a pancreatic cancer animal disease model in which the expression level of SCG5 protein or SCG5 mRNA in the blood is increased; and
(b) 단계 (a) 이후 상기 동물 모델로부터 혈액을 채취하고, 채취된 혈액 내 SCG5 단백질 또는 SCG5 mRNA 발현 수준을 측정하는 단계; 채취된 혈액 내 SCG5 단백질 또는 SCG5 mRNA 발현 수준이 의약 후보 물질을 투여에 의해 유의한 증가를 보이는 경우, 상기 의약 후보 물질은 췌장암 치료제로 판정한다.(b) collecting blood from the animal model after step (a) and measuring the expression level of SCG5 protein or SCG5 mRNA in the collected blood; If the expression level of SCG5 protein or SCG5 mRNA in the collected blood shows a significant increase due to administration of the drug candidate, the drug candidate is determined to be a treatment for pancreatic cancer.
본 발명의 단계 (a)의 투여는 경구 또는 비경구로 투여 가능하며, 의약 후보 물질의 특성에 따라, 당업자가 적합한 투여 경로를 선택할 수 있다. Step (a) of the present invention can be administered orally or parenterally, and depending on the characteristics of the drug candidate, a person skilled in the art can select an appropriate administration route.
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(a) 본 발명은 췌장암 진단용 조성물을 제공한다. (a) The present invention provides a composition for diagnosing pancreatic cancer.
(b) 본 발명은 췌장암 진단 키트를 제공한다.(b) The present invention provides a pancreatic cancer diagnostic kit.
(c) 본 발명은 췌장암 진단 방법을 제공한다. (c) The present invention provides a method for diagnosing pancreatic cancer.
(d) 본 발명은 췌장암 치료제 스크리닝 방법을 제공한다.(d) The present invention provides a screening method for pancreatic cancer treatment.
(e) 본 발명의 췌장암 진단용 조성물 또는 진단 키트를 활용하면 혈액 검사를 통해 매우 신속하고 간편하게 췌장암을 조기 진단할 수 있다. (e) Using the composition or diagnostic kit for pancreatic cancer of the present invention, pancreatic cancer can be diagnosed very quickly and easily through a blood test.
도 1은 분석 흐름도를 나타낸다.
도 2는 기계학습 선별 진단마커후보(feature) 별 ROC curve 기반 성능평가 결과를 나타낸다.
도 3은 기계학습 선정 6개 진단마커후보(feature)의 ROC curve를 통한 평가 결과를 나타낸다.
도 4는 기계학습 선정 6개 진단마커후보(feature)의 ROC curve를 통한 평가 결과 (정상 vs 췌장암 1기)를 나타낸다.
도 5는 정상 또는 비암 췌장 조직과 췌장암 조직에서 SCG5 유전자 발현 정도의 비교 결과를 나타낸다.
도 6은 정상 또는 비암 췌장 조직과 췌장암 조직에서 SCG5 유전자 발현 정도의 비교 결과를 나타낸다.
도 7은 남성(좌)과 여성(우)의 정상 또는 비암 췌장 조직과 췌장암 조직에서 SCG5 유전자 발현 정도의 비교 결과를 나타낸다.
도 8은 정상 또는 비암 췌장 조직과 췌장암 조직에서 SCG5 유전자 발현과 체질량지수(BMI)와의 상관관계 분석 결과를 나타낸다.
도 9는 조직(tissue) 별 SCG5 발현 수준을 나타낸다. Figure 1 shows the analysis flow diagram.
Figure 2 shows the ROC curve-based performance evaluation results for each machine learning screening diagnostic marker candidate (feature).
Figure 3 shows the evaluation results through ROC curve of six diagnostic marker candidates (features) selected by machine learning.
Figure 4 shows the evaluation results (normal vs. stage 1 pancreatic cancer) of the six diagnostic marker candidates (features) selected by machine learning through ROC curve.
Figure 5 shows the results of comparison of SCG5 gene expression levels in normal or non-cancerous pancreatic tissue and pancreatic cancer tissue.
Figure 6 shows the results of comparison of SCG5 gene expression levels in normal or non-cancerous pancreatic tissue and pancreatic cancer tissue.
Figure 7 shows the comparison results of SCG5 gene expression levels in normal or non-cancerous pancreatic tissue and pancreatic cancer tissue of men (left) and women (right).
Figure 8 shows the results of correlation analysis between SCG5 gene expression and body mass index (BMI) in normal or non-cancerous pancreatic tissue and pancreatic cancer tissue.
Figure 9 shows the SCG5 expression level by tissue.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
실시예Example
실시예 1: 분석 흐름도Example 1: Analysis flow chart
NCBI GEO 데이터베이스에서 제공하는 췌장암 관련 데이터셋 3개, GSE16515, GSE62165, GSE71729를 이용하여 분석을 진행하였다. GSE16515는 29,626개의 마이크로어레이 칩 프로브, 16개의 정상 샘플, 36개의 췌장암 샘플을 포함하고, GSE62165는 48,784개의 마이크로어레이 칩 프로브, 13개의 정상 샘플, 118개의 췌장암 샘플을 포함한다. 그리고 GSE71729는 19,749개의 마이크로어레이 칩 프로브, 46개의 정상 샘플, 145개의 췌장암 샘플을 포함한다. 이 분석의 목적이 혈액분비단백질을 코딩하는 유전자의 발현을 분석함으로 췌장암 진단 혈중 마커를 찾기 위함이었으므로 Human Protein Atlas에서 제공하는 1708개의 혈액분비체유전자로 우선 데이터 전처리를 하고 이후 분석을 진행하였는데, 데이터 전처리 후 얻은 유전자의 개수는 GSE16515에서 2,679개(프로브 중복 포함), GSE62165에서 3,498개(프로브 중복 포함), GSE71729에서 1,449개였다.Analysis was conducted using three pancreatic cancer-related datasets, GSE16515, GSE62165, and GSE71729, provided by the NCBI GEO database. GSE16515 contains 29,626 microarray chip probes, 16 normal samples, and 36 pancreatic cancer samples, and GSE62165 contains 48,784 microarray chip probes, 13 normal samples, and 118 pancreatic cancer samples. And GSE71729 includes 19,749 microarray chip probes, 46 normal samples, and 145 pancreatic cancer samples. Since the purpose of this analysis was to find diagnostic blood markers for pancreatic cancer by analyzing the expression of genes encoding blood secretion proteins, data were first preprocessed with 1708 blood secretion genes provided by the Human Protein Atlas and then analyzed. The number of genes obtained after preprocessing was 2,679 in GSE16515 (including probe duplication), 3,498 in GSE62165 (including probe duplication), and 1,449 in GSE71729.
정상 또는 비암 췌장조직의 전사체와 췌장암 전사체의 발현과 통계적 유의차를 log2 fold-change는 1 이상, -1 이하, p-value는 0.05 이하로 계산하고 Volcano plot으로 표현하여 평가하였다. 이로부터 얻은 유의발현 유전자(Differentially Expressed Genes)는 GSE16515에서 410개(프로브 중복 포함), GSE62165에서 930개(프로브 중복 포함), GSE71729에서 144개이었고, 혈액에서 검출하기 용이해야하므로 유전자 발현 값을 특정값 이상의 발현값을 갖는 유전자로 선별했다. 선별된 유전자는 총 26개였고, 이를 가지고 췌장암 발병유무를 분별할 수 있는 모델을 확립하기 위해 랜덤포레스트 알고리즘 이용하여 머신러닝을 시행하였다. 그 결과, 10개의 feature를 가진 모델이 가장 AUC가 높게 나왔고, 정확도가 가장 높은 것은 6개의 feature를 가진 모델이었다(참조: 도 1).The expression and statistical significant difference between the transcriptome of normal or non-cancerous pancreatic tissue and the pancreatic cancer transcript were evaluated by calculating the log2 fold-change as 1 or more, -1 or less, and p-value less than 0.05 and expressing them in a Volcano plot. The significantly expressed genes obtained from this were 410 in GSE16515 (including probe duplication), 930 in GSE62165 (including probe duplication), and 144 in GSE71729. Since it should be easy to detect in blood, the gene expression value was specified. Genes with expression values above this value were selected. There were a total of 26 selected genes, and machine learning was performed using the random forest algorithm to establish a model that could distinguish the presence or absence of pancreatic cancer. As a result, the model with 10 features had the highest AUC, and the model with 6 features had the highest accuracy (see Figure 1).
상기 분석을 통해 선별된 26개의 유전자를 이용하여 기계학습을 통해 얻은 모델의 평가 결과를 표로 나타내었다(표 1). 진단 모델의 성능 평가를 위해 그린 ROC curve에서 10개의 진단마커후보(feature)를 가진 모델의 AUROC가 0.9027 (90.3%)로 가장 높았고(표 2), 6개의 진단마커후보(feature)를 가진 모델의 정확도(accuracy)가 0.8919 (89.2%)로 가장 높았다(표 3).The evaluation results of the model obtained through machine learning using the 26 genes selected through the above analysis are presented in a table (Table 1). In the ROC curve drawn to evaluate the performance of the diagnostic model, the AUROC of the model with 10 diagnostic marker candidates (features) was the highest at 0.9027 (90.3%) (Table 2), and the model with 6 diagnostic marker candidates (features) had the highest AUROC of 0.9027 (90.3%). Accuracy was the highest at 0.8919 (89.2%) (Table 3).
아래 표 2 및 3의 경우, 기계학습 기반 분석에 의해 10개 또는 6개의 진단마커후보(feature)로 선별된 유전자 리스트를 나타낸다.Tables 2 and 3 below show a list of genes selected as 10 or 6 diagnostic marker candidates (features) by machine learning-based analysis.
실시예 2: 기계학습 선정 진단마커후보(feature) 별 ROC curve 기반 성능평가Example 2: Performance evaluation based on ROC curve for each machine learning selected diagnostic marker candidate (feature)
기계학습을 통해 도출한 진단 모델의 성능평가를 위해 민감도와 특이도를 바탕으로 분류/예측 모델의 유용성 측정 방법인 ROC 곡선(수신자 조작 특성 곡선)을 그렸고, ROC 곡선의 아래 면적(area under the ROC, AUROC)으로 나타내어 비교하였다. 선별한 유전자 26개를 모두 포함한 기계학습 선정 모델의 AUROC 값은 0.891, 선별된 유전자 10개, 8개, 6개, 4개, 2개를 포함한 기계학습 선정 모델은 AUROC 값은 각각 0.903, 0.887, 0.859, 0.865, 0.858이었다(참조: 도 2). 따라서 AUROC 값을 기준으로는 유전자 10개를 포함하는 기계학습 모델이 가장 높은 값을 나타내었다. To evaluate the performance of the diagnostic model derived through machine learning, an ROC curve (receiver operating characteristic curve), a method of measuring the usefulness of the classification/prediction model, was drawn based on sensitivity and specificity, and the area under the ROC curve was drawn. , AUROC) and compared. The AUROC value of the machine learning selection model including all 26 selected genes was 0.891, and the AUROC value of the machine learning selection model including 10, 8, 6, 4, and 2 selected genes was 0.903, 0.887, respectively. They were 0.859, 0.865, and 0.858 (see Figure 2). Therefore, based on the AUROC value, the machine learning model including 10 genes showed the highest value.
실시예 3: 기계학습 선정 6개 진단마커후보(feature)의 ROC curve를 통한 평가 결과Example 3: Evaluation results through ROC curve of six diagnostic marker candidates (features) selected by machine learning
정확도(accuracy)가 가장 높은 진단 모델에 속한 6개의 유전자의 ROC curve를 그려 진단능의 결과를 나타내었다. PRSS3의 AUROC는 0.769, 민감도(Sensitivity)는 80.3%, 특이도(Specificity)는 74.7%, 양성예측도(Positive Predictive Value)는92.7%, 음성예측도(Negative Predictive Value)는 48.7%였다. CTRB2는 AUROC 0.78, 민감도 80.3%, 특이도 74.7%, 양성예측도 92.7%, 음성예측도 48.7%였고, SPARC는 AUROC 0.696, 민감도 88.3%, 특이도 53.3%, 양성예측도 88.3%, 음성예측도 53.3%였다. CLU는 AUROC 0.715, 민감도 83.9%, 특이도 50.7%, 양성예측도 87.2%, 음성예측도 44.2%였고, MMP7은 AUROC 0.663, 민감도 74.2%, 특이도 56.0%, 양성예측도 87.1%, 음성예측도 35.3%였다. 그리고 SCG5는 AUROC 0.690, 민감도 75.3%, 특이도 60.0%, 양성예측도 88.2%, 음성예측도 37.8%였다(참조: 도 3). 요약하면, 10개의 feature(유전자)를 가진 모델이 가장 AUROC가 높게 나왔고, 정확도가 가장 높은 것은 6개의 feature(유전자)를 가진 모델이었다 (참조: 도 1).The ROC curve of the six genes belonging to the diagnostic model with the highest accuracy was drawn to show the results of diagnostic ability. The AUROC of PRSS3 was 0.769, sensitivity was 80.3%, specificity was 74.7%, positive predictive value was 92.7%, and negative predictive value was 48.7%. CTRB2 had AUROC 0.78, sensitivity 80.3%, specificity 74.7%, positive predictive value 92.7%, and negative predictive value 48.7%, and SPARC had AUROC 0.696, sensitivity 88.3%, specificity 53.3%, positive predictive value 88.3%, and negative predictive value. It was 53.3%. CLU had AUROC 0.715, sensitivity 83.9%, specificity 50.7%, positive predictive value 87.2%, and negative predictive value 44.2%, and MMP7 had AUROC 0.663, sensitivity 74.2%, specificity 56.0%, positive predictive value 87.1%, and negative predictive value. It was 35.3%. And SCG5 had AUROC of 0.690, sensitivity of 75.3%, specificity of 60.0%, positive predictive value of 88.2%, and negative predictive value of 37.8% (see Figure 3). In summary, the model with 10 features (genes) had the highest AUROC, and the model with 6 features (genes) had the highest accuracy (see Figure 1).
실시예 4: 기계학습 선정 6개 진단마커후보(feature)의 ROC curve를 통한 평가 결과 (정상 vs 췌장암 1기)Example 4: Evaluation results through ROC curve of 6 diagnostic marker candidates (features) selected by machine learning (normal vs. stage 1 pancreatic cancer)
기계학습을 통해 선정한 진단모델에 속한 6개의 유전자를 가지고 정상 샘플과 췌장암 1기 샘플에서 ROC curve를 그려 진단능의 결과를 나타냈다. PRSS3은 AUROC 0.981, 민감도 100.0%, 특이도 87.5%, 양성예측도 86.7%, 음성예측도 100.0%, CTRB2는 AUROC 0.837, 민감도 100.0%, 특이도 75.0%, 양성예측도 86.7%, 음성예측도 100.0%, SPARC는 AUROC 1.000, 민감도 100.0%, 특이도 100.0%, 양성예측도 100.0%, 음성예측도 100.0%, CLU는 AUROC 0.981, 민감도 100.0%, 특이도 87.5%, 양성예측도 92.9%, 음성예측도 100.0%, MMP7는 AUROC 0.827, 민감도 100.0%, 특이도 75.0%, 양성예측도 86.7%, 음성예측도 100.0%, SCG5는 AUROC 0.788, 민감도 84.6%, 특이도 62.5%, 양성예측도 78.6%, 음성예측도 74.4%로 계산됐다(참조: 도 4). Using 6 genes belonging to the diagnostic model selected through machine learning, ROC curves were drawn in normal samples and stage 1 pancreatic cancer samples to show the results of diagnostic ability. PRSS3 has AUROC 0.981, sensitivity 100.0%, specificity 87.5%, positive predictive value 86.7%, negative predictive value 100.0%, CTRB2 has AUROC 0.837, sensitivity 100.0%, specificity 75.0%, positive predictive value 86.7%, negative predictive value 100.0. %, SPARC has AUROC 1.000, sensitivity 100.0%, specificity 100.0%, positive predictive value 100.0%, negative predictive value 100.0%, CLU has AUROC 0.981, sensitivity 100.0%, specificity 87.5%, positive predictive value 92.9%, negative predictive value. Figure 100.0%, MMP7 has AUROC 0.827, sensitivity 100.0%, specificity 75.0%, positive predictive value 86.7%, negative predictive value 100.0%, SCG5 has AUROC 0.788, sensitivity 84.6%, specificity 62.5%, positive predictive value 78.6%, Negative prediction was also calculated to be 74.4% (see Figure 4).
실시예 5: 정상 또는 비암 췌장 조직과 췌장암 조직에서 SCG5 유전자 발현 정도의 비교Example 5: Comparison of SCG5 gene expression levels in normal or non-cancerous pancreatic tissue and pancreatic cancer tissue
정상 또는 비암 췌장 조직과 초기 췌장암(stage I) 조직에서 SCG5의 유전자 발현 정도를 비교하기 위해 boxplot을 그려 분석을 시행하였다. GSE62165 데이터에서 13개의 정상 또는 비암 조직 샘플, 8개의 초기 췌장암 (stage I) 조직 샘플을 가지고 분석을 진행하였는데, 먼저 해당 데이터의 정규성 여부를 먼저 확인하기 위해 Shapiro-Wilk test(샤피로 윌크 검정)를 시행하였고, p-value = 0.637로서 정규분포임을 확인하였다. 정상 또는 비암 조직 샘플과 초기 췌장암 샘플 간 SCG5 발현 농도 차이를 확인하기 위해 Student’s t-test를 시행하였고, 그 결과 p-value = 0.03으로 정상 또는 비암 조직 샘플과 초기 췌장암 조직 샘플 간 SCG5 발현 농도가 유의미한 차이를 보이는 것을 확인하였다(참조: 도 5).Analysis was performed by drawing a boxplot to compare the gene expression level of SCG5 in normal or non-cancerous pancreatic tissue and early pancreatic cancer (stage I) tissue. In the GSE62165 data, analysis was conducted with 13 normal or non-cancerous tissue samples and 8 early pancreatic cancer (stage I) tissue samples. First, the Shapiro-Wilk test was performed to check whether the data was normal. and p-value = 0.637, confirming normal distribution. Student's t-test was performed to determine the difference in SCG5 expression concentration between normal or non-cancerous tissue samples and early pancreatic cancer samples. As a result, p-value = 0.03, indicating that the SCG5 expression concentration between normal or non-cancerous tissue samples and early pancreatic cancer tissue samples was significant. It was confirmed that there was a difference (see Figure 5).
실시예 6: 췌장암 환자와 비교통제군(비암)의 혈중 SCG5 수준 비교Example 6: Comparison of SCG5 levels in the blood of pancreatic cancer patients and comparison control group (non-cancer)
ELISA를 이용해 췌장암 환자 25인 및 비교통제군(비암) 25인의 혈중 SCG5의 발현농도를 측정하고 췌장암 환자 샘플과 정상 샘플을 비교하였다. ELISA kit (SCG5 ELISA kit, MyBioSource, INC. 미국)를 이용한 혈중 SCG5 분석은 제조사의 지시에 준하여 실시하였다. 췌장암 환자와 비교통제군(비암)의 혈장을 희석하고 96-웰 플레이트에서 1차, 2차 항체 반응 후, 결합된 2차 항체의 검출은 기질용액을 사용하여 발색을 실시하였다. 기질 용액과의 반응을 수행하고 ELISA 판독기(Microplate reader, SpectraMax 190, Molecular Device, 미국)를 이용하여 흡광도 450nm에서 OD 값을 측정하였다. 먼저 해당 데이터의 정규성 여부를 확인하기 위해 shapiro test를 시행하였고, p-value = 0.595로서 정규분포임을 확인하였다. SCG5 발현농도의 그룹간 차이를 확인하기 위해 Student’s t-test를 이용하였고, 그 결과 p-value = 4.9e-06로 정상 샘플과 췌장암 환자 샘플 간 SCG5 발현농도에 유의미한 차이가 있음을 확인하였다. 또한 SCG5의 발현농도를 이용하여 췌장암 환자 샘플과 정상 샘플의 예측 가능성을 확인하기 위해 ROC curve를 계산하였다. 그 결과 민감도(Sensitivity) 92.0%, 특이도(Specificity)는 68.0%를 나타냈고, 진단정확도(AUROC)는 85.9%, 두 그룹을 구분지을 수 있는 최적의 농도값은 106.27 ng/ml로 계산되었다(참조: 도 6).Using ELISA, we measured the expression level of SCG5 in the blood of 25 pancreatic cancer patients and 25 control groups (non-cancer) and compared pancreatic cancer patient samples with normal samples. Blood SCG5 analysis was performed using an ELISA kit (SCG5 ELISA kit, MyBioSource, INC. USA) according to the manufacturer's instructions. Plasma from pancreatic cancer patients and comparative controls (non-cancer) was diluted and reacted with primary and secondary antibodies in a 96-well plate. The bound secondary antibodies were detected using color development using a substrate solution. The reaction with the substrate solution was performed and the OD value was measured at absorbance 450 nm using an ELISA reader (Microplate reader, SpectraMax 190, Molecular Device, USA). First, a shapiro test was performed to check whether the data was normal, and p-value = 0.595, confirming normal distribution. Student's t-test was used to confirm the difference between groups in SCG5 expression level, and the result was p-value = 4.9e-06, confirming that there was a significant difference in SCG5 expression level between normal samples and pancreatic cancer patient samples. In addition, an ROC curve was calculated to confirm the predictability of pancreatic cancer patient samples and normal samples using the expression level of SCG5. As a result, sensitivity was 92.0%, specificity was 68.0%, diagnostic accuracy (AUROC) was 85.9%, and the optimal concentration value for distinguishing between the two groups was calculated to be 106.27 ng/ml ( See also: Figure 6).
실시예 7: 남성과 여성의 정상 또는 비암 췌장 조직과 췌장암 조직에서 SCG5 유전자 발현 정도의 비교Example 7: Comparison of SCG5 gene expression levels in normal or non-cancerous pancreatic tissue and pancreatic cancer tissue of men and women
정상인과 췌장암 환자의 SCG5 발현 분석이 성별 간 어떤 차이가 있는지 확인하기 위해 남성과 여성으로 데이터를 나눠 분석을 진행하였다. 남성(좌), 여성(우) 데이터 모두 정규성을 먼저 확인해 정규분포임을 확인하였다(남성 p-value = 0.959, 여성 p-value = 0.162). 정상인 혈액 샘플과 췌장암 환자 혈액 샘플 내 SCG5의 발현 차이는 Student’s t-test로 계산했는데, 남성 데이터는 p-value=0.0027, 여성 데이터는 p-value=0.00027로 남성, 여성 데이터 모두 유의미한 차이를 보였지만, 여성 데이터에서 정상인 혈액 샘플과 췌장암 환자 혈액 샘플 간 SCG5 발현량 차이가 좀 더 명확함을 확인할 수 있었다(참조: 도 7).To determine whether there were any differences between genders in the SCG5 expression analysis of normal subjects and pancreatic cancer patients, the data was divided into men and women and analyzed. The normality of both male (left) and female (right) data was first checked to confirm that they were normally distributed (male p-value = 0.959, female p-value = 0.162). The difference in expression of SCG5 between normal blood samples and pancreatic cancer patient blood samples was calculated using Student's t-test. Male data showed p-value=0.0027 and female data showed p-value=0.00027, showing significant differences in both male and female data. In the female data, it was confirmed that the difference in SCG5 expression level between normal blood samples and blood samples of pancreatic cancer patients was more clear (see Figure 7).
실시예 8: 정상 또는 비암 췌장 조직과 췌장암 조직에서 SCG5 유전자 발현과 체질량지수(BMI)와의 상관관계 분석Example 8: Correlation analysis between SCG5 gene expression and body mass index (BMI) in normal or non-cancerous pancreatic tissue and pancreatic cancer tissue
정상인과 췌장암 환자의 BMI값이 SCG5의 발현과 어떤 관계가 있는지 확인하기 위해 상관 분석을 진행했다. 정상인 중 남성은 13명, 여성은 12명이고, BMI(kg/m2)의 범위는 19.60 ~ 32.46 이었으며, SCG5의 발현 범위는 28.50 ~ 216.91 (ng/ml)이었다. 췌장암 환자 중 남성은 15명, 여성은 10명이고, BMI((kg/m2)의 범위는 16.68 ~ 26.90이었으며, SCG5의 발현 범위는 7.40 ~ 143.90 (ng/ml)이었다. BMI와 SCG5 사이에는 피어슨 상관 계수는 0.44, 스피어만 상관 계수는 0.39로 양의 상관관계가 있음을 확인했다(각각 p-value = 0.0016, p-value = 0.0059)(참조: 도 8).A correlation analysis was performed to determine the relationship between the BMI values of normal people and pancreatic cancer patients with the expression of SCG5. Among the normal subjects, there were 13 men and 12 women, the range of BMI (kg/m 2 ) was 19.60 to 32.46, and the expression range of SCG5 was 28.50 to 216.91 (ng/ml). Among pancreatic cancer patients, there were 15 men and 10 women. The range of BMI (kg/m 2 ) was 16.68 to 26.90, and the expression range of SCG5 was 7.40 to 143.90 (ng/ml). There was a difference between BMI and SCG5. The Pearson correlation coefficient was 0.44 and the Spearman correlation coefficient was 0.39, confirming a positive correlation (p-value = 0.0016 and p-value = 0.0059, respectively) (see Figure 8).
실시예 9: 조직(tissue) 별 SCG5 발현 수준Example 9: SCG5 expression level by tissue
정상 공여자에서 조직(tissue)별로 RNA-seq 분석을 수행하여 생성된 공공데이터베이스 Genotype-Tissue Expression (GTEx)에서 조직 별 SCG5 유전자 발현 수준을 그래프로 나타내었다. 뇌하수체(Pituitary gland)를 제외하고는 췌장조직이 특이적으로 SCG5발현을 높게 보이는 것을 알 수 있었다. 이는, 췌장암 환자에서 감소되는 SCG5 혈중 양이 췌장암 특이적 진단 역할을 할 수 있음을 확인해주는 결과이다(참조: 도 9).The SCG5 gene expression level for each tissue was graphed in the public database Genotype-Tissue Expression (GTEx), which was created by performing RNA-seq analysis for each tissue in normal donors. It was found that, except for the pituitary gland, pancreatic tissue specifically showed high SCG5 expression. This is a result confirming that the reduced blood amount of SCG5 in pancreatic cancer patients can play a role in pancreatic cancer-specific diagnosis (see Figure 9).
Claims (9)
A composition for diagnosing stage 1 pancreatic cancer, comprising an agent for detecting the mRNA expression level of the SCG5 protein or the gene encoding the SCG5 protein in a blood sample isolated from the human body.
The composition for diagnosing stage 1 pancreatic cancer according to claim 1, wherein the human body is a woman.
The method of claim 1, wherein the agent for detecting the SCG5 protein level is at least one selected from the group consisting of an antibody, aptamer, ligand, PNA (peptide nucleic acid), and oligopeptide that specifically binds to the SCG5 protein. Composition for diagnosing stage 1 pancreatic cancer.
The method of claim 1, wherein the agent for detecting the mRNA expression level of the gene encoding the SCG5 protein is at least one selected from the group consisting of a primer, a probe, and an antisense nucleotide that specifically binds to the mRNA, stage 1 pancreatic cancer. Diagnostic composition.
A kit for diagnosing stage 1 pancreatic cancer, comprising the composition according to any one of claims 1 to 4.
(a) 인체로부터 분리된 혈액 시료 내의 SCG5 단백질 또는 SCG5 단백질을 암호화하는 유전자의 mRNA 발현 수준을 측정하는 단계; 정상 혈액 대조군 시료에 비해 상기 SCG5 단백질 수준 또는 상기 mRNA 발현 수준이 감소된 경우, 제1기 췌장암으로 진단함.
Method for diagnosing stage 1 pancreatic cancer, including the following steps:
(a) measuring the mRNA expression level of the SCG5 protein or the gene encoding the SCG5 protein in a blood sample isolated from the human body; When the SCG5 protein level or the mRNA expression level is decreased compared to a normal blood control sample, stage 1 pancreatic cancer is diagnosed.
The method of claim 6, wherein the step of measuring the SCG5 protein expression level is performed using SCG5, which is at least one selected from the group consisting of an antibody, aptamer, ligand, PNA (peptide nucleic acid), and oligopeptide that specifically binds to the SCG5 protein. A method for diagnosing stage 1 pancreatic cancer, which is performed using an agent that detects protein levels.
The method of claim 6, wherein the step of measuring the mRNA expression level of the gene encoding the SCG5 protein detects the mRNA expression level of at least one selected from the group consisting of a primer, a probe, and an antisense nucleotide that specifically binds to the mRNA. A method for diagnosing stage 1 pancreatic cancer, which is performed using an agent.
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