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Application method of circular RNA CircSERPINE2 and detection and treatment preparation

An application method and preparation technology, applied in the field of tumor molecular biology, can solve the problems of poor treatment effect of ovarian tumors, no opportunity for surgery, and rapid tumor spread.

Active Publication Date: 2021-07-20
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Platinum and paclitaxel are less effective in treating recurrent ovarian tumors, and recurrent tumors spread rapidly, resulting in no opportunity for surgery and no treatment options for patients

Method used

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  • Application method of circular RNA CircSERPINE2 and detection and treatment preparation
  • Application method of circular RNA CircSERPINE2 and detection and treatment preparation
  • Application method of circular RNA CircSERPINE2 and detection and treatment preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 CircSERPINE2 detection kit in ovarian cancer cells A2780 and OVCAR3 and their cisplatin- and carboplatin-resistant cells

[0062] 1. Isopropanol 100ml

[0063] 2. Trizol reagent 100ml

[0064] 3. Chloroform 50ml

[0065] 4.1 μM random reverse transcription primer 50 μl

[0066] 5. Enzyme-free water 2ml

[0067] 6. 10mMdNTP100μl

[0068] 7. 200U / μl RNA reverse transcriptase 50μl

[0069] 8.5× reverse transcription buffer 1ml

[0070] 9.40U / μl RNA inhibitor 500μl

[0071] 10. Premix Ex Taq 50μl

[0072] 11. 10 μM lncRNAENST00000529841 specific primer 50 μl

[0073] Specific primers for fluorescent quantitative PCR:

[0074] CircSERPINE2 forward primer: 5'-GGGTCTGTGGAAATCACGGT-3'

[0075] CircSERPINE2 reverse primer: 5'-CAAGGACGACCACACCGGAA-3'

[0076] 12. 50 μl each of 10 μM internal reference and control primers

[0077] Specific primers for internal reference β-actin:

[0078] 5'-TCACCAACTGGGACGACATG-3'

[0079] 5'-GTCACCGGAGTCCATCCGAT-3';

[00...

Embodiment 2

[0086] Detection of CircSERPINE2 in Ovarian Cancer Cells and Drug-resistant Ovarian Cancer Cells

[0087] (1) Collect ovarian cancer cells A2780 and OVCAR3 and their induced cisplatin- and carboplatin-resistant cells, and extract total RNA: deplete the medium in the culture plate after induction, add 1ml Trizol, and place horizontally at room temperature for 5 minutes, Make the lysate evenly distributed on 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 200 μl chloroform, cover the centrifuge tube cap tightly, shake up and down for 15 seconds, and let it stand at room temperature. Place for 3 minutes, centrifuge at 12,000g, 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 transfer the light-colored supernatant to another centrifuge tube, ...

Embodiment 3

[0118] Example 3 Real-time fluorescent quantitative PCR analysis of differential expression of CircSERPINE2 in different ovarian cancer cell lines and drug-resistant tissues

[0119] The IOSE80, A2780, OVCAR3, SKOV3 and HO8910 used were purchased from the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences. The above several types of cells were cultured to a cell density of 80%, and then the method of extracting RNA, reverse transcription and QPCR to detect the expression level of CircSERPINE2 was the same as in Examples 1 and 2. The detection results showed that CircSERPINE2 was significantly highly expressed in the drug-resistant cell lines OVCAR3 and SKOV3, and was low-expressed in human normal ovarian epithelial cells IOSE80 and sensitive cell line A2780. Among them, HO8910 is a cell line related to tumor invasion and metastasis. For the results, see image 3 .

[0120] The present invention collected 4 cases of platinum-resistant (R-1, 2, 3, 4) and 3...

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Abstract

The invention discloses an application method of a circular RNA CircSERPINE2 and a detection and treatment preparation. The invention relates to a detection reagent for detecting ovarian cancer platinum chemotherapy tolerance, and in particular relates 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. Researches prove that the CircSERPINE2 is highly expressed in the platinum-resistant ovarian cancer tissue, and the CircSERPINE2 is a marker of the platinum resistance of the ovarian cancer, so that the CircSERPINE2 expression 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 CircSERPINE2 expression has profound clinical significance and generalization performance.

Description

technical field [0001] The invention relates to the field of tumor molecular biology, in particular to an application method, detection and treatment preparation of circular RNA CircSERPINE2. Background technique [0002] In 2018, approximately 22,240 new cases of ovarian cancer were diagnosed in the United States, with approximately 14,070 deaths from ovarian cancer, according to the most recent ovarian cancer statistics. The incidence of ovarian cancer accounts for only 2.5% of all gynecological malignancies, but its mortality rate accounts for 5% of all gynecological cancer mortality. The main treatments for ovarian cancer are surgery and chemotherapy. Conventional chemotherapy is a combination of platinum and paclitaxel as first-line chemotherapy, 15%-25% of patients are resistant to the main treatment, and the remaining 70-85% of patients are sensitive to the first chemotherapy. After initial treatment, tumors shrink significantly in size, but most tumors recur within...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/686C12N15/113
CPCC12Q1/6886C12Q1/686C12Q2600/106C12Q2600/158C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 彭淑平帅词俊莫雨晴钟雁城刘映贺甜甜邹姿
Owner CENT SOUTH UNIV
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