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Micro RNA related to BafilomycinA1 liver cancer and ovarian cancer inhibition cell line

A technology of ovarian cancer cells and liver cancer cell lines, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problem of no similar inventions being found, and achieve the effect of eliminating genetic differences

Inactive Publication Date: 2015-05-20
JIANGSU UNIV
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  • Claims
  • Application Information

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Problems solved by technology

Through domestic and foreign literature search, no similar inventions were found

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  • Micro RNA related to BafilomycinA1 liver cancer and ovarian cancer inhibition cell line
  • Micro RNA related to BafilomycinA1 liver cancer and ovarian cancer inhibition cell line
  • Micro RNA related to BafilomycinA1 liver cancer and ovarian cancer inhibition cell line

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Experimental program
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Effect test

specific Embodiment approach        Embodiment 1

[0023] Inhibitory effect of BafilomycinA1 on liver cancer cell line BEL-7402 and ovarian cancer cell line HO-8910

[0024] (1) Detection of BafilomycinA1 on liver cancer cell line BEL-7402 and ovarian cancer cell line HO-8910 by WST-1 kit

[0025] The liver cancer cell line BEL-7402 and the ovarian cancer cell line HO-8910 were purchased from the Cell Bank of the Chinese Academy of Sciences and stored in a medium containing 10% fetal bovine serum (product of Hangzhou Sijiqing Bioengineering Materials Co., Ltd.), 100 U / mL penicillin and 100 RPMI-1640 of μg / mL streptomycin (Life technologies, GIBCO TM product) at 37 °C, 5% CO 2 cultivated under conditions. Use the WST-1 Cell Proliferation Kit (Shanghai Biyuntian Biotechnology Co., Ltd.) to detect the cell proliferation inhibition test of Bafilomycin A1 on the above two types of cells: Add 200 μL to each well of a 96-well culture plate at a concentration of 5×10 4 / mL cell suspension, cultured for 20h to allow the cells to ad...

Embodiment 2

[0036] High-throughput analysis and qPCR verification of microRNAs related to the inhibition of BafilomycinA1 on liver cancer cell line BEL-7402 and ovarian cancer cell line HO-8910.

[0037] After 400nM Bafilomycin A1 acted on BEL-7402 and HO-8910 for 48h, the total cellular RNA of the drug-dosed group and the control group were treated with Trizol (Life technologies, Invitrogen TM Product) extraction, measure purity and quality with NanoDrop ND-1000 spectrophotometer, accord with OD 260 / OD 280 > 1.6, OD 260 / OD 230 > 1 and RIN value (RNA Integrity Number) > 5, carry out reverse transcription, labeling, hybridization, elution and signal processing, the operation process is strictly in accordance with the instructions, the membrane is Affymetrix GeneChip Human Gene 1.0 ST Array, and a Group microRNA, as shown in Table 1.

[0038] The operation steps of microRNA qPCR verification of BEL-7402 and HO-8910 co-sensitivity to Bafilomycin A1 are as follows: reverse transcript...

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Abstract

The invention relates to a group of micro RNA related to a BafilomycinA1 liver cancer and ovarian cancer inhibition cell line, belonging to the technical field of medical biology. The preparation method comprises the following steps: respectively acting BafilomycinA1 on a liver cancer cell line BEL-7402 and an ovarian cancer cell line HO-8910, and detecting the in-vitro growth inhibition, apoptosis promoting and invasion inhibition effects of BafilomycinA1 on two cells by performing soft agar cloning formation assay, electron microscopy observation, apoptosis related enzyme detection and in-vitro invasion assay; respectively acting 400nM BafilomycinA1 on the liver cancer cell line BEL-7402 and the ovarian cancer cell line HO-8910 for 48 hours, collecting dosing cells and control cells, extracting the RNA, analyzing the high-flux micro RNA between the drug group and the control group by using Affymetrix GeneChip Human Gene Array micro RNA array, and verifying the differences of the micro RNA by using qPCR, thereby obtaining co-sensitive micro RNA of the two cells. According to the group of micro RNA sensitive to the BafilomycinA1, on one hand, the micro RNA has the cancer inhibition effectiveness corresponding to the BafilomycinA1; on the other hand, the micro RNA has obvious change in two tumor cells, and a basis is provided for targets designed by taking the micro RNA as a potential broad spectrum anti-cancer drug.

Description

technical field [0001] The invention relates to a group of microRNA related to BafilomycinA1 inhibiting liver cancer and ovarian cancer cell lines, and belongs to the field of medical biotechnology. Background technique [0002] BafilomycinA1 is a specific inhibitor of hydrogen ion pump V-ATPase (Marshansky V, et al. [0003] Eukaryotic V-ATPase: novel structural findings and functional insights. Biochimica et biophysica acta 1837: 857-879, 2014.). The hydrogen ion pump of tumor cells is often overactive, which means that it can promote the excess acid produced by the Wurgerg effect to discharge cells, maintain the weak alkalinity of cells, and avoid apoptosis induced by acidosis; promote acidification of tumor extracellular matrix and increase protease activity , thereby enhancing the invasiveness of tumor cells; related to the generation of drug resistance of tumor cells; affecting the transmission of cell signals. As a specific inhibitor of V-ATPase, BafilomycinA1 can i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113
Inventor 卢小东
Owner JIANGSU UNIV
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