Application method of long-chain non-coding RNA LOC107987064 and detection and treatment preparation

A RNALOC107987064, long-chain non-coding technology, applied in the field of molecular biology, can solve the problems of insufficient responsiveness of platinum-based chemotherapy regimens and no major breakthroughs, and achieve far-reaching practical significance and promotional effects

Active Publication Date: 2021-08-10
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ovarian cancer cell debulking surgery and platinum-paclitaxel combination chemotherapy are the standard treatment options for patients with advanced ovarian cancer, but more than 25% of patients in the early stage will develop resistance to platinum-based chemotherapy, that is, within 6 months after stopping chemotherapy Relapsed, and less than 30% of partially resistant patients responded to subsequent platinum-based chemotherapy regimens
Research on novel biomarkers associated with platinum resistance in ovarian cancer has been ongoing, but there has been no major breakthrough in the study of a series of molecular changes involving genes, transcription, proteins, and metabolism

Method used

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  • Application method of long-chain non-coding RNA LOC107987064 and detection and treatment preparation
  • Application method of long-chain non-coding RNA LOC107987064 and detection and treatment preparation
  • Application method of long-chain non-coding RNA LOC107987064 and detection and treatment preparation

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Experimental program
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Effect test

Embodiment 1

[0091] Detection Kit for LncRNA LOC107987064 in Tumor Tissues from Human Ovarian Cancer Patients

[0092] 1. Isopropanol 100ml

[0093] 2. Trizol reagent 100ml

[0094] 3. Chloroform 50ml

[0095] 4. 1μM random reverse transcription primer 50μl

[0096] 5. Enzyme-free water 2ml

[0097] 6. 10mM dNTP 100μl

[0098] 7. 200U / μl RNA reverse transcriptase 50μl

[0099] 8. 5× reverse transcription buffer 1ml

[0100] 9. 40U / μl RNase inhibitor 500μl

[0101] 10.Premix Ex Taq 50μl

[0102] 11. 10μM lncRNA LOC107987064 specific primer 50μl

[0103] Specific primers for fluorescent quantitative PCR:

[0104] lncRNA LOC107987064 forward primer: 5'-TCAGTGAGGGCCCCAGTTAG-3'

[0105] lncRNA LOC107987064 reverse primer: 5'-CTTTCAAGCTGGACAGAGCAT-3'

[0106] 12. 50 μl each of 10 μM internal reference and control primers

[0107] Specific primers for internal reference β-actin:

[0108] 5'-CCTATCGAGCATGGAGTGGT-3'

[0109] 5'-CTGAGGCATAGAGGGACAGC-3'

[0110] Specific primers for in...

Embodiment 2

[0115] In the previous research work, the applicant collected 8 tumor tissue samples from platinum-resistant and platinum-sensitive ovarian cancer patients, extracted total RNA from each tumor tissue sample, and performed real-time fluorescent quantitative PCR to analyze the expression of LncRNALOC107987064 after reverse transcription, and found that: The expression of LncRNA LOC107987064 was higher in tumor tissues of platinum-resistant patients than in platinum-sensitive patients (see image 3), and has a statistical difference, which is also consistent with the trend of high-throughput sequencing in the previous period. Accordingly, the applicant proposed to use LncRNA LOC107987064 to prepare a marker detection reagent for platinum resistance in human ovarian cancer. According to the results of repeated detection of LOC107987064 expression in platinum-resistant and platinum-sensitive patient tissues by qPCR experiments, we concluded that when the internal reference Actin cyc...

Embodiment 3

[0117] Detection of LncRNA LOC107987064 in Human Ovarian Cancer Cells and Induced Ovarian Cancer Platinum-Resistant Cells

[0118] (1) Collect human ovarian cancer parental cells and induced platinum-resistant ovarian cancer cells, and extract total RNA: Remove the culture medium in the culture plate, add 1ml Trizol, and place it horizontally at room temperature for 5 minutes, so that the lysate is evenly distributed in the Cell surface, then blow the cells with a pipette gun to make the cells fall off; transfer the cell lysate to a centrifuge tube, add 200μl chloroform, close the cap of the centrifuge tube tightly, shake up and down for 15 seconds, let stand on ice for 5 minutes, 12000g , centrifuge at 4°C for 15 minutes. Take out the centrifuge tube, the sample is divided into three layers: the light-colored supernatant aqueous phase, the middle white layer and the pink lower organic phase. Carefully absorb the light-colored supernatant and transfer it to another centrifuge...

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Abstract

The invention discloses an application method of long-chain non-coding RNA LOC107987064, a detection preparation and a treatment preparation. The invention relates to a detection reagent for detecting ovarian cancer platinum chemotherapy tolerance, in particular to a kit for detecting whether ovarian cancer is tolerant to platinum drugs or not by a real-time fluorescence quantitative analysis method and a human ovarian cancer platinum tolerance treatment preparation. The researches prove that the LncRNA LOC107987064 is highly expressed in a platinum-resistant ovarian cancer tissue, and the LOC107987064 is a marker of the platinum resistance of the ovarian cancer, so that the expression of the LOC107987064 is applied to the detection of the platinum chemotherapy resistance of the ovarian cancer and the enhancement of the chemotherapy sensitivity of the human ovarian cancer platinum drugs through inhibition, and the LncRNA LOC107987064 has profound clinical significance and generalization performance.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to an application method, detection and treatment preparation of long-chain non-coding RNA LOC107987064. Background technique [0002] Ovarian cancer is a highly malignant disease. Because of its insidious onset and lack of specific clinical symptoms in the early stage, most of them are already in clinical stage III or IV when they are discovered. Ovarian cancer cell debulking surgery and platinum-paclitaxel combination chemotherapy are the standard treatment options for patients with advanced ovarian cancer, but more than 25% of patients in the early stage will develop resistance to platinum-based chemotherapy, that is, within 6 months after stopping chemotherapy Less than 30% of patients who have relapsed and are partially resistant respond to subsequent platinum-based chemotherapy regimens. Research on novel biomarkers associated with platinum resistance in ovarian cancer has be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/113A61K45/00A61P35/00
CPCC12Q1/6886C12Q1/6851C12N15/113A61K45/00A61P35/00C12Q2600/106C12Q2600/158C12Q2600/178C12N2310/14C12N2310/531C12Q2531/113C12Q2563/107C12Q2561/113
Inventor 彭淑平帅词俊钟雁城刘映贺甜甜莫雨晴邹姿
Owner CENT SOUTH UNIV
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