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Application method of long-chain non-coding RNA LOC730101 and detection and treatment preparation

A long-chain non-coding, application-method technology, applied in DNA/RNA fragments, medical preparations containing active ingredients, recombinant DNA technology, etc. Profound practical significance and promotion effect

Active Publication Date: 2021-06-18
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 LOC730101 and detection and treatment preparation
  • Application method of long-chain non-coding RNA LOC730101 and detection and treatment preparation
  • Application method of long-chain non-coding RNA LOC730101 and detection and treatment preparation

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Embodiment 1

[0052] Detection Kit for LncRNA LOC730101 in Tumor Tissues from Human Ovarian Cancer Patients

[0053] 1. Isopropanol 100ml

[0054] 2. Trizol reagent 100ml

[0055] 3. Chloroform 50ml

[0056] 4. 1μM random reverse transcription primer 50μl

[0057] 5. Enzyme-free water 2ml

[0058] 6. 10mM dNTP 100μl

[0059] 7. 200U / μl RNA reverse transcriptase 50μl

[0060] 8. 5× reverse transcription buffer 1ml

[0061] 9. 40U / μl RNase inhibitor 500μl

[0062] 10.Premix Ex Taq 50μl

[0063] 11. 10μM lncRNA LOC730101 specific primer 50μl

[0064] Specific primers for fluorescent quantitative PCR:

[0065] lncRNA LOC730101 forward primer: 5'-TCTCACCCTACCGTTCGTCT-3'

[0066] lncRNA LOC730101 reverse primer: 5'-AGTGGCTTTGGGAGTTCAGG-3'

[0067] 12. 50 μl each of 10 μM internal reference and control primers

[0068] Specific primers for internal reference β-actin:

[0069] 5'-CCTATCGAGCATGGAGTGGT-3'

[0070] 5'-CTGAGGCATAGAGGGACAGC-3'

[0071] Specific primers for internal refere...

Embodiment 2

[0077] In the previous research work, the applicant collected 8 tumor tissue samples from platinum-resistant and platinum-sensitive patients with ovarian cancer, extracted total RNA from each tumor tissue sample, and performed real-time fluorescent quantitative PCR to analyze the expression of LncRNA LOC730101 after reverse transcription, and found that : The expression of LncRNA LOC730101 is higher in tumor tissues of platinum-sensitive patients than in platinum-resistant patients (see figure 2 ), and there is a statistical difference, which is also consistent with the trend of high-throughput sequencing in the previous period. Accordingly, the applicant proposes to use LncRNALOC730101 to prepare a marker detection reagent sensitive to platinum therapy in human ovarian cancer. According to the results of repeated detection of LOC730101 expression in platinum-resistant and platinum-sensitive patient tissues by qPCR experiments, we concluded that when the internal reference Act...

Embodiment 3

[0079] Detection of LncRNA LOC730101 in Human Ovarian Cancer Cells and Induced Ovarian Cancer Platinum-Resistant Cells

[0080] (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 On the surface of the cells, blow the cells with a pipette gun to make the cells fall off; transfer the cell lysate to a centrifuge tube, add 200 μl of 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...

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Abstract

The invention discloses an application method of long-chain non-coding RNA LOC730101 and a detection and treatment preparation. The invention relates to a detection reagent for detecting the sensitivity of ovarian cancer platinum chemotherapy, in particular to a kit for detecting whether ovarian cancer is sensitive to platinum drugs or not by a real-time fluorescence quantitative analysis method and a human ovarian cancer platinum tolerance treatment preparation. Researches prove that the LncRNA LOC730101 is highly expressed in platinum-sensitive ovarian cancer tissues, and the LOC730101 is an ovarian cancer platinum-sensitive marker, so that the expression of the LOC730101 is applied to the detection of the ovarian cancer platinum chemosensitivity and the enhancement of the chemosensitivity of human ovarian cancer platinum drugs through overexpression, and 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 LOC730101. 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 been ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11A61K31/7088A61P35/00
CPCC12Q1/6886C12Q1/6851A61K31/7088A61P35/00C12Q2600/106C12Q2600/158C12Q2600/166C12Q2600/178C12Q2531/113C12Q2563/107C12Q2545/101
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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