CN114958856B - Application of long-chain non-coding RNA CYP1B1-AS1 AS breast cancer biomarker and treatment target - Google Patents
Application of long-chain non-coding RNA CYP1B1-AS1 AS breast cancer biomarker and treatment target Download PDFInfo
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- CN114958856B CN114958856B CN202210733639.5A CN202210733639A CN114958856B CN 114958856 B CN114958856 B CN 114958856B CN 202210733639 A CN202210733639 A CN 202210733639A CN 114958856 B CN114958856 B CN 114958856B
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Abstract
The invention discloses long-chain non-coding RNA CYP1B1-AS1, the nucleotide sequence of which is shown AS SEQ ID NO. 1. The invention also discloses a CYP1B1-AS1 lentivirus over-expression vector, a lentivirus stable expression cell strain and application thereof in preparing a breast cancer diagnosis kit, a medicament for preventing or treating breast cancer or a prognosis product. CYP1B1-AS1 can obviously down-regulate expression in breast cancer, and can be used AS a diagnosis marker of the breast cancer. The expression level of CYP1B1-AS1 is related to prognosis of patients, and can be used AS a prognosis marker of breast cancer. Up-regulating CYP1B1-AS1 can inhibit malignant progress of breast cancer cells, and has application value AS a gene therapy target. Upregulation of CYP1B1-AS1 expression can increase sensitivity of breast cancer cells to chemotherapeutic agents. The invention can be used for the preparation of products such as diagnosis and treatment related to breast cancer, improvement of targeted drug sensitivity and the like, and has a certain clinical transformation prospect.
Description
Technical Field
The invention belongs to the technical field of tumor molecular biology, and relates to application of long-chain non-coding RNA CYP1B1-AS1 AS a breast cancer diagnosis and prognosis biomarker and a treatment target.
Background
Breast cancer is the first malignancy in global incidence and is the leading cause of cancer death in women. The incidence rate of breast cancer in China is continuously increased, and the final treatment failure of patients is often caused by lack of timely and effective diagnosis and treatment. Therefore, the development mechanism of breast cancer is deeply explored, a new diagnosis and treatment target is searched, the application value is mined, and the clinical transformation potential is unprecedented.
Recent research results show that long non-coding RNA (lncRNA) with the length of more than 200nt can interact with DNA, RNA or protein, participate in important regulation and control such as chromosome modification, transcription, nuclear transport, translation, post-translational modification and the like, act on various cell signal transduction pathways and participate in the whole process of tumor progression. Deregulation of LncRNA expression is one of the characteristic changes of the human cancer transcriptome, possessing the potential to be a tumor biomarker and therapeutic target molecule.
The invention relies on Jiangsu province tumor hospitals, and the applicant bears a plurality of national and provincial natural science foundation projects related to tumors, and accumulates rich research experience of tumor related molecular markers. In recent years, genetic engineering techniques have rapidly progressed in tumor research, and genetic diagnosis and molecular targeted therapy have shown great advantages in the clinic of various malignant tumors. Therefore, the pathogenesis of the breast cancer is deeply known, the effect of the related molecular markers in the disease development is clear, and a foundation can be laid for the individual and accurate diagnosis and treatment of the breast cancer.
Disclosure of Invention
The invention aims to: the technical problem to be solved by the invention is to provide long-chain non-coding RNA CYP1B1-AS1 beneficial to breast cancer diagnosis and treatment, and the application potential of the lncRNA CYP1B1-AS1 in solving the clinical situation of breast cancer is disclosed.
The invention also solves the technical problem of providing an application of CYP1B1-AS1 lentivirus over-expression vector and lentivirus stable expression cell strain in preparing a breast cancer diagnosis kit, a medicament for preventing or treating breast cancer or a prognosis product.
The invention also solves the technical problem of providing a fluorescent quantitative PCR detection kit.
The technical scheme is as follows: in order to solve the technical problems, the invention provides long-chain non-coding RNA CYP1B1-AS1, and the nucleotide sequence of the long-chain non-coding RNA CYP1B1-AS1 is shown AS SEQ ID NO. 1.
The CYP1B1-AS1 gene is all called: cytochrome P450 family 1 subfamily B member 1-anti RNA 1, located on chromosome 2 sense strand, is 1776nt in length and carries a polyA tail.
Seq ID NO.1
＞NR_027252.1Homo sapiens CYP1B1 antisense RNA 1(CYP1B1-AS1)，long non-coding RNA
The invention also comprises biological functions of CYP1B1-AS1 in breast cancer cells, and discloses application value of the CYP1B1-AS1 serving AS a molecular therapeutic target.
The invention also comprises a CYP1B1-AS1 lentiviral overexpression vector, wherein the CYP1B1-AS1 lentiviral overexpression vector contains the long-chain non-coding RNA CYP1B1-AS1.
The invention also comprises a lentivirus stable expression cell strain, wherein the lentivirus stable expression cell strain is obtained by packaging the CYP1B1-AS1 lentivirus over-expression vector into virus particles to infect host cells.
Wherein the host cell is an MCF7 or MDA-MB-231 cell.
The invention up-regulates the expression of CYP1B1-AS1 in breast cancer cell strains MCF7 and MDA-MB-231 through a lentivirus over-expression vector, and confirms that the up-regulates CYP1B1-AS1 inhibits the malignant progress of breast cancer cells through a cell proliferation experiment, a clone formation experiment, a flow cell cycle detection and the like.
The invention also discloses application of the long-chain non-coding RNA CYP1B1-AS1 serving AS a breast cancer diagnosis marker.
The invention also comprises the application of the long-chain non-coding RNA CYP1B1-AS1, the CYP1B1-AS1 lentivirus over-expression vector and the lentivirus stable expression cell strain in preparing a breast cancer diagnosis kit, a medicament for preventing or treating breast cancer or a prognosis product.
The invention also discloses a fluorescent quantitative PCR detection kit, which comprises a specific primer pair, wherein the primer pair sequences of the specific primer pair are SEQ ID NO.2 and SEQ ID NO.3.
The detection kit also comprises a reverse transcription reagent and a real-time fluorescent quantitative PCR reagent, and a preparation for detecting the CYP1B1-AS1 expression level in breast cancer tissues by using the real-time fluorescent quantitative PCR.
The detection kit further comprises an internal reference GAPDH primer pair, and the sequence of the internal reference GAPDH primer pair is shown as SEQ ID NO.4 and SEQ ID NO. 5.
The CYP1B1-AS1 detection primers used for real-time fluorescence quantitative PCR are AS follows:
CYP1B1-AS1：
SEQ ID NO.2：Forward Primer(5’to3’)
SEQ ID NO.3：Forward Primer(5’to3’)
internal reference GAPDH:
SEQ ID NO.4：Forward Primer(5’to3’)
SEQ ID NO.5：Forward Primer(5’to3’)
the invention also discloses application of the long-chain non-coding RNA CYP1B1-AS1, the CYP1B1-AS1 lentivirus over-expression vector and the lentivirus stable expression cell strain in preparing a product for improving sensitivity of breast cancer cells to chemotherapy.
The CYP1B1-AS1 lentiviral overexpression vector is LV 6-homoCYP 1B1-AS1 recombinant plasmid, and is constructed by Shanghai Ji Ma pharmaceutical technology Co.
The sequences of the primers used for constructing the CYP1B1-AS1 lentiviral overexpression vector plus the cleavage sites NotI and NsiI are AS follows:
SEQ ID NO.6：Forward Primer(5’to3’)
SEQ ID NO.7：Forward Primer(5’to3’)
the invention also comprises a cell strain which is obtained by packaging the lentiviral recombinant vector LV 6-homoCYP 1B1-AS1 and then infecting cells to obtain stable expression.
The invention utilizes the slow virus over-expression vector to up-regulate CYP1B1-AS1, which is beneficial to improving the sensitivity of breast cancer cells to common chemotherapeutic drugs cisplatin.
Wherein the application treats breast cancer cells by adding different concentrations of cisplatin, and then determining the influence of CYP1B1-AS1 on the chemotherapeutic drug sensitivity of the cancer cells by adding CCK-8.
Wherein the concentration of cisplatin is 25 μg/mL,12.5 μg/mL,6.25 μg/mL,3.13 μg/mL,1.56 μg/mL,0.78 μg/mL.
The beneficial effects are that: compared with the prior art, the invention has the following advantages:
1. the invention discovers that CYP1B1-AS1 can obviously down-regulate expression in breast cancer for the first time, and can be used AS a diagnosis marker of the breast cancer.
2. The invention discloses that the expression level of CYP1B1-AS1 is related to prognosis of patients for the first time, and can be used AS a prognosis marker of breast cancer.
3. The invention definitely up-regulates CYP1B1-AS1 for the first time, can inhibit malignant progress of breast cancer cells, and has application value AS a gene therapy target.
4. The invention proves for the first time that up-regulating the expression of CYP1B1-AS1 can improve the sensitivity of breast cancer cells to chemotherapeutics.
In conclusion, the invention can be used for the preparation of products such as diagnosis, treatment, targeted drug sensitivity improvement and the like related to breast cancer, and has a certain clinical transformation prospect.
Drawings
FIG. 1 thermal analysis of detection results of lncRNA chip of breast cancer tissue. And (3) injection: t, tumor tissue, N, paracancerous normal tissue.
FIG. 2TCGA data analyze CYP1B1-AS1 expression in breast cancer.
FIG. 3NCBI database shows structural features of CYP1B1-AS1.
FIG. 4LNCipedia database analyzes CYP1B1-AS1 encoding ability and conservation.
FIG. 5 is a graph showing the evaluation of the value of CYP1B1-AS1 detection in combination with clinical breast cancer tissue samples.
FIG. 6 is a graph showing the relationship between CYP1B1-AS1 expression level and prognosis of breast cancer patients.
FIG. 7 biological function study of CYP1B1-AS1. A. qPCR detects the effect of up-regulating expression of CYP1B1-AS1 in breast cancer cells. B. The CCK-8 assay detects the effect of upregulation of CYP1B1-AS1 on proliferation capacity of breast cancer cells. C. The clonogenic assay detects the effect of upregulation of CYP1B1-AS1 on the clonogenic capacity of breast cancer cells. D. Flow cytometric analysis up-regulates the effects of CYP1B1-AS1 on breast cancer cell cycle. And (3) injection: breast cancer cells that up-regulate CYP1B1-AS1 expression and negative controls thereof are labeled MCF7-exp, MCF7-NC and MDA-231-exp, MDA-231-NC, respectively.
FIG. 8 effects of over-expression of CYP1B1-AS1 on drug sensitivity of breast cancer cells.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
EXAMPLE 1 lncRNA chip detection of differentially expressed lncRNA-CYP1B1-AS1 in breast cancer tissue
1. Experimental materials and methods
The ethical committee approved that 6 breast cancer tissues and paracancerous normal tissues were selected from the biological sample library, and the expression profile of lncRNA and mRNA in the tissues was detected using an Arraystar human lncRNA chip, which was completed by Shanghai health biosystems. The chip was able to detect 40,173 lncRNA and 20,730 protein-encoding transcripts. Through analysis by Genespring GX v12.1 software and through Quantile standardization, the differential expression lncRNAs or differential expression mRNAs with statistical significance between two groups of samples are screened by Fold change more than or equal to 2.0, and P is less than 0.05.
2. Experimental results
In the embodiment, by taking a paracancerous normal tissue AS a control, fold change is more than or equal to 2, P is less than 0.05 AS a screening index, CYP1B1-AS1 is found to be significantly down-regulated in breast cancer tissues, and GEPIA2 analysis of cancer genome map (the Cancer Genome Atlas, TCGA) data shows that: CYP1B1-AS1 down-regulates expression in breast cancer (FIGS. 1, 2). CYP1B1-AS1 is an antisense lncRNA with a polyA tail, located on the sense strand of chromosome 2, 1776nt in length and carrying a polyA tail. CYP1B1-AS1 does not possess the ability to encode proteins (FIGS. 3, 4).
Example 2 application of CYP1B1-AS1 detection in preparation of Breast cancer diagnosis and prognosis products
1. Experimental materials and methods
1.1 Breast cancer tissue sample collection
30 cases of primary breast cancer and paracancerous normal tissue samples were collected, approved by the ethical committee, and immediately preserved with liquid nitrogen until RNA extraction.
1.2RNA extraction, reverse transcription and qPCR reactions
Extraction Using Trizol reagent (Invitrogen)Total RNA in cells, total number of cells of single extracted RNA is not less than 10 6 And OD is used 260/280 Detection of RNA purity, when OD 260/280 Between 1.9 and 2.1, can be used for subsequent detection.
cDNA was synthesized using 1. Mu.g total RNA using PrimeScript RT kit (TaKaRa) and stored at-20℃for later use. The primers for CYP1B1-AS1 are AS follows: an upstream primer: 5'-ACTGGTATGACCACCCGAGA-3'; a downstream primer: 5'-ACTGGCATACTGGTCACTGC-3', tm=60℃. The primers for the internal reference GAPDH were as follows, the upstream primer: 5'-ATGGGTGTGAACCATGAGAA-3'; a downstream primer: 5'-GTGCTAAGCAGTTGGTGGTG-3', tm=60℃. Primers were synthesized by Shanghai Biotechnology.
Real-time fluorescent quantitative PCR detection and melting curve analysis were performed using a LightCycler PCR apparatus from Roche, USA, using TB from TakaraPremix Ex Taq TM II, performing PCR reaction by using a kit, amplifying 40 cycles by using a two-step method, collecting fluorescent signals, analyzing an amplification curve and a melting curve by using self-contained software of an instrument, calculating a cycle threshold (Ct), and using 2 -ΔΔCt The expression level of the relative gene is obtained by calculation.
The specific operation and reaction system are as follows:
1.2.1RNA extraction
(1) Trizol was added to the tissue samples at 50-100 mg/mL. The cells were completely lysed and transferred to a centrifuge tube after being homogenized thoroughly with an electric homogenizer for about 1-2 min.
(2) Into the centrifuge tube containing the lysate, 0.2mL of chloroform (0.2 mL of chloroform was added to 1mL of Trizol) was added, and the mixture was thoroughly mixed by shaking on a shaker for 20s and left at room temperature for 5min.
(3) Centrifugation at 12000rpm for 10min at 4℃and then pipetting the total RNA-containing upper aqueous phase into a new centrifuge tube, about 0.5mL of upper aqueous phase per mL of Trizol. The uptake of DNA and proteins from the organic phase and the middle layer should be avoided.
(4) Adding isopropanol with the same volume as the upper water phase, reversing for mixing for several times, and precipitating at room temperature for 5min. Centrifuge at 12000rpm at 4℃for 15min.
(5) The supernatant was discarded, 1mL of 75% ethanol was added per mL of Trizol, and the mixture was gently inverted and mixed to wash the RNA pellet twice. Centrifuging at 12000rpm and 4 ℃ for 2min, discarding the liquid, and reversely airing at room temperature.
(6) An appropriate amount of DEPC-treated water was added to dissolve the RNA precipitate. Stored at-80 ℃.
1.2.2 reverse transcription:
(1) removal of genomic DNA
Reaction conditions: 42 ℃,2min,4 ℃ and preserving.
(2) Reverse transcription reaction:
the reaction product was added to the following reagents:
the reaction condition is 37 ℃ for 15min;85 ℃,5s;4 ℃, and preserving.
1.2.3PCR amplification:
the reverse transcription cDNA was aspirated at 2. Mu.L and the following reaction system was added:
1.3Kaplan-Meier Plotter analysis of CYP1B1-AS1 expression versus prognosis in breast cancer patients.
1.4 data processing and analysis
Wilcoxon signed rank test was used to compare breast cancer tissue with paracancerous normal tissue samples. Statistical analysis was performed using GraphPad Prism 8.0 software. P values less than 0.05 are considered statistically significant (×p < 0.05, ×p < 0.01, ×p < 0.001).
2. Experimental results
In this example, CYP1B1-AS1 in 30 breast cancer tissues and paracancerous normal tissues was detected by RT-PCR, and CYP1B1-AS1 in the cancer tissues was found to be significantly lower than that in the paracancerous normal tissues, and the subject operating profile (receiver operating characteristic curve, ROC) showed an area under the curve (area under the ROC curve, AUC) of 0.909, [95% confidence interval: 0.839-0.979, P < 0.001], suggesting that CYP1B1-AS1 detection has higher diagnostic value (FIG. 5).
In addition, kaplan-Meier Plotter analysis showed that: the high recurrence-free survival Rate (RFS) and the high total survival rate (OS) of the patients with the breast cancer with the CYP1B1-AS1 expression show that the application value of the patients with the breast cancer is used AS a clinical prognosis factor of the breast cancer (figure 6).
Example 3 biological Functions of CYP1B1-AS1 in breast cancer cells
1. Experimental materials and methods
1.1 cell culture
Breast cancer cell lines MCF7 and MDA-MB-231 were purchased from the Shanghai cell bank of the national academy of sciences and cultured according to ATCC protocol: MCF7 cells were cultured in RPMI 1640 containing 10% fetal bovine serum and incubated at 37℃with 5% CO 2 In an incubator. MDA-MB-231 cells were cultured using L-15 containing 10% FBS, and placed in an air incubator at 37 ℃.293T cells were purchased from Shanghai cell Bank, china academy of sciences and cultured with DMEM containing 10% FBS.
1.2 establishment of lentiviral Stable expression cell lines
A stable expression cell line of CYP1B1-AS1 was established by using a lentiviral overexpression vector (LV 6, shanghai Ji Ma pharmaceutical technologies Co., ltd.): the sequences of the primers for amplifying the target gene homoCYP1B1-AS1 plus the cleavage sites NotI and NsiI are AS follows: an upstream primer: 5'-AGGGTTCCAAGCTTAAGCGGCCGCCAACTATGGTTGCTAACACAAGAATCGGCA-3', downstream primer: 5'-GATCCATCCCTAGGTAGATGCATTTTTTTTTTTTTTTTTTTTTTTTTTATTAAACTG-3', tm=55 ℃; cloning a target gene Homo CYP1B1-AS1 into a lentiviral overexpression vector (LV 6, shanghai Ji Ma pharmaceutical technology Co., ltd.) to obtain an LV6-Homo CYP1B1-AS1 recombinant plasmid (H2470-2), transfecting 293T cells with the lentiviral overexpression vector LV6-Homo CYP1B1-AS1 recombinant plasmid and the lentiviral overexpression vector LV6 (negative control) respectively by using liposome Lipofectamine 3000 (Invitrogen) for virus packaging, collecting virus liquid in cell culture supernatant after 72 hours, respectively infecting MCF7 and MDA-MB-231 cells, adding 10 mug/mL of puromycin for screening after 48 hours, and obtaining MCF7 and MDA-MB-231 cells stably expressing CYP1B1-AS1 and negative controls thereof (marked AS MCF7-exp, MCF 7-MDA-231-exp and MDA-231-NC respectively). The construction and packaging of lentiviral expression vectors was completed by Shanghai Ji Ma pharmaceutical technologies. The effect of over-expression of CYP1B1-AS1 was examined using the RT-PCR method in example 2, and the results are shown in FIG. 7A.
1.3 cell proliferation assay
The proliferation potency of MCF7-exp, MCF7-NC and MDA-231-exp, MDA-231-NC cells was examined using CCK-8. The treated test cells were spread in 96-well plates (3X 10 per well) 3 And 2) were incubated for 2 hours with 10% CCK-8 in the medium at 0h, 24h,48h,72h and 96h, respectively, and absorbance was measured at a wavelength of 450 nm. CCK-8 was purchased from the Japan same Coulter chemical institute.
1.4 cloning experiments
MCF7-exp, MCF7-NC and MDA-231-exp, MDA-231-NC cells were seeded in six-well plates containing 2mL of RPMI 1640 (MCF 7) or L-15 (MDA-MB-231) culture solution at a gradient density of 300, 500, 800 cells, respectively, and gently rotated to disperse the cells uniformly. Culturing in a cell culture incubator for 1-2 weeks. When macroscopic clones appeared in the dishes, the supernatant was discarded and washed 2 times with PBS. Cells were fixed by adding 5mL of 4% paraformaldehyde for 15min. Removing the fixing solution, adding a proper amount of crystal violet staining solution for dyeing for 20min, then slowly washing the staining solution with flowing water, drying, and observing the cell clone formation condition under a microscope.
1.5 cell cycle detection
Cell cycle analysis was performed using propidium iodide staining: MCF7-exp, MCF7-NC and MDA-231-exp, MDA-231-NC cells were digested with trypsin, washed with PBS, and fixed with 70% ethanol at-20 ℃. After centrifugation, the cells were incubated in a hypotonic solution containing 50. Mu.g/mL propidium iodide, 0.1% sodium citrate, 0.1% Triton X-100 and 20. Mu.g/mL for 30min at 37 ℃. The cells were then analyzed in linear mode using a flow cytometer.
1.6 data processing and analysis
t-test and analysis of variance (analysis of variance, ANOVA) were used for the inter-group comparison. Statistical analysis was performed using GraphPad Prism 8.0 software. P values less than 0.05 are considered statistically significant (×p < 0.05, ×p < 0.01).
2. Experimental results
The lentiviral overexpression vector in this example can stably up-regulate CYP1B1-AS1 expression in breast cancer cells MCF7 and MDA-MB-231. Cell proliferation experiments, clonogenic experiments and flow-through cell cycle assays showed that upregulation of CYP1B1-AS1 expression was effective in inhibiting cell proliferation (FIG. 7).
Example 5 application of CYP1B1-AS1 in preparation of products for increasing sensitivity of breast cancer cells to chemotherapy
1. Experimental materials and methods
1.1 drug sensitivity test
MCF7-exp, MCF7-NC and MDA-231-exp, MDA-231-NC cells were used at 5X 10 per well 3 Cisplatin (dissolved in physiological saline) was added to the cells at various concentrations (25. Mu.g/mL, 12.5. Mu.g/mL, 6.25. Mu.g/mL, 3.13. Mu.g/mL, 1.56. Mu.g/mL, 0.78. Mu.g/mL) after incubation for 24 h. An equal volume of solvent was added as a control group with drug concentration of 0. After 48h of treatment, the culture broth was replaced with 100. Mu.L of a reaction broth containing 10% CCK-8 (10% CCK-8 was prepared in RPMI 1640 or L-15 broth, respectively, as-prepared), absorbance was measured at a wavelength of 450nm after incubation for 3h, and a dose-response curve was drawn.
1.2 data processing and analysis
ANOVA was used for group-to-group comparisons and statistical analysis was performed using GraphPad Prism8 software. P values less than 0.05 are considered statistically significant (×p < 0.05, ×p < 0.01).
2. Experimental results
In this example we treated breast cancer cells with different concentrations of cisplatin and the activity of the cells was measured by adding CCK-8, which indicated that up-regulation of CYP1B1-AS1 expression was beneficial in increasing the sensitivity of breast cancer cells to chemotherapeutic drugs (figure 8).
Sequence listing
<110> Jiangsu province tumor Hospital
<120> application of long-chain non-coding RNA CYP1B1-AS1 AS breast cancer biomarker and treatment target
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ttgacttgag cggcaaagga cccccatagt caggctgtga tttgaggagc atcaagaggc 1500
cagagtttcc aagttgggac ccagaaggtg gggttggcag ggggagggta ggagcagctg 1560
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Claims (5)
- The application of the CYP1B1-AS1 lentiviral overexpression vector in preparing a medicament for treating breast cancer is characterized in that the CYP1B1-AS1 lentiviral overexpression vector contains long-chain non-coding RNA CYP1B1-AS1, and the nucleotide sequence of the long-chain non-coding RNA CYP1B1-AS1 is shown AS SEQ ID NO. 1.
- 2. The application of a lentivirus stable expression cell strain in preparing a medicament for treating breast cancer is that the lentivirus stable expression cell strain is obtained by packaging a CYP1B1-AS1 lentivirus over-expression vector into virus particle infected cells, wherein the CYP1B1-AS1 lentivirus over-expression vector contains long-chain non-coding RNA CYP1B1-AS1, and the nucleotide sequence of the long-chain non-coding RNA CYP1B1-AS1 is shown AS SEQ ID NO. 1.
- 3. The application of long-chain non-coding RNA CYP1B1-AS1 in preparing a product for improving the sensitivity of breast cancer cells to a chemotherapeutic drug is characterized in that the nucleotide sequence of the long-chain non-coding RNA CYP1B1-AS1 is shown AS SEQ ID NO.1, and the chemotherapeutic drug is cisplatin.
- The application of CYP1B1-AS1 lentiviral overexpression vector in preparation of a product for improving sensitivity of breast cancer cells to chemotherapeutics is characterized in that the CYP1B1-AS1 lentiviral overexpression vector contains long-chain non-coding RNA CYP1B1-AS1, the nucleotide sequence of the long-chain non-coding RNA CYP1B1-AS1 is shown AS SEQ ID NO.1, and the chemotherapeutics is cisplatin.
- 5. The application of a lentiviral stable expression cell strain in preparing a product for improving sensitivity of breast cancer cells to a chemotherapeutic drug is that the lentiviral stable expression cell strain is obtained by packaging a CYP1B1-AS1 lentiviral over-expression vector into virus particle infected cells, wherein the CYP1B1-AS1 lentiviral over-expression vector contains long-chain non-coding RNA CYP1B1-AS1, the nucleotide sequence of the long-chain non-coding RNA CYP1B1-AS1 is shown AS SEQ ID NO.1, and the chemotherapeutic drug is cisplatin.
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| CN114627970A (en) * | 2022-03-15 | 2022-06-14 | 中南大学湘雅医院 | Prognosis model of scorching-related lncRNA of colon adenocarcinoma and construction method and application thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114627970A (en) * | 2022-03-15 | 2022-06-14 | 中南大学湘雅医院 | Prognosis model of scorching-related lncRNA of colon adenocarcinoma and construction method and application thereof |
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| Homo sapiens CYP1B1 antisense RNA 1 (CYP1B1-AS1), long non-coding RNA;GenBank Accession：NR_027252.1;《GenBank》 * |
| Landscape of tumor suppressor long noncoding RNAs in breast cancer;Boran Pang等;《Journal of Experimental & Clinical Cancer Research》;第38卷;文章编号:79 * |
| LncRNA CARMN overexpression promotes prognosis and chemosensitivity of triple negative breast cancer via acting as miR143-3p host gene and inhibiting DNA replication;Xiaonan Sheng等;《Journal of Experimental & Clinical Cancer Research》;第40卷;文章编号: 205 * |
| LncRNAs and miRNAs participate in determination of sensitivity of cancer cells to cisplatin;Taheri M等;《Experimental and Molecular Pathology》;第123卷;文章编号:104602 * |
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