Oophoroma multidrug resistance cell strain established by etoposide induction

A multi-drug resistance and cell line technology, applied in the field of tumor biology, can solve the problems of easy drug resistance and affecting the degree of drug resistance of tumor cells

Inactive Publication Date: 2009-08-12
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies at home and abroad have compared these two methods, and it is believed that different induction methods may obtain drug-resistant cell lines with different drug resistance mechanisms, and may also affect the degree of drug resistance of tumor cells. Continuous induction is more likely to produce drug resistance than shock induction

Method used

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  • Oophoroma multidrug resistance cell strain established by etoposide induction
  • Oophoroma multidrug resistance cell strain established by etoposide induction
  • Oophoroma multidrug resistance cell strain established by etoposide induction

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] SKOV3 / VP16 multidrug-resistant cell line is established according to the following steps: (1) human ovarian cancer SKOV3 cell line, after recovery, use DMEM culture fluid (containing 10% calf serum, penicillin 100U / ml, streptomycin 100ug / ml) At 37°C, 5% CO 2 , 95% humidity conditions; (2) Take the SKOV3 cells in the logarithmic growth phase, change the fresh culture medium, add VP16 to make the concentration of 0.1 μg / ml, at 37 ° C, 5% CO 2 , continue to cultivate under 95% humidity conditions, change the medium in time, and maintain the drug concentration of 0.1 μg / ml until the surviving cells can grow stably and pass on at this concentration; (3) Appropriately increase the concentration of VP16. 2 , Continue culturing under 95% humidity conditions, change the medium in time, and maintain the drug concentration until the surviving cells can grow stably and pass on at this concentration; (4) Repeat step (3) until the stable growth, passage and recovery in 3 μg / ml VP16 a...

Embodiment 2

[0012] The SKOV3 / VP16 cell strain that this method is established carries out morphological observation and biological characteristic identification:

[0013] 1. Morphological observation

[0014] (1) Inverted phase-contrast microscope to observe the morphology of living cells

[0015] Take a bottle of SKOV3 and SKOV3 / VP16 cells in the logarithmic growth phase, and observe the morphology of the living cells under an inverted phase-contrast microscope after changing the medium and take pictures. It can be seen that the SKOV3 cells are polygonal, the shape is more consistent, and the intracellular structure is uniform (as shown in Figure 1); the size and shape of the SKOV3 / VP16 cells are different, and there are many dark substances in the cells, especially near the cell membrane (such as Figure 2).

[0016] (2) HE staining to observe cell morphology

[0017] Take SKOV3 and SKOV3 / VP16 cells in the logarithmic growth phase, digest with 0.25% trypsin to make a single cell suspe...

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Abstract

The invention relates to a multidrug resistant cell strain for oophoroma, which is characterized in that a human oophoroma SKOV3 cell strain is selected as a parental cell; the action concentration of chemotherapeutics of etoposide (VP16) is gradually increased from 0.1 mu g/ml to 3 mu g/ml; and drug resistant SKOV3/VP16 cell strain is established. The SKOV3/VP16 cell can stably grow, transfer and reanimate in 3 mu g/ml VP16, makes the VP16 generate drug resistant index (RI) of 21.548, and performs intersect drug on MTX, DOX, DDP, VCR, 5-Fu and MMC except the VP16. The invention aims to provide a drug resistant tumor cell model for relative study on the tumor.

Description

technical field [0001] The invention belongs to the field of tumor biology and relates to a drug-resistant human ovarian cancer cell line. Specifically, it involves using human ovarian cancer SKOV3 as a parental cell, and establishing the multidrug-resistant ovarian cancer cell line SKOV3 / VP16 by gradually increasing the concentration of the chemotherapeutic drug etoposide (VP16). Background technique [0002] Among the malignant tumors of the female reproductive system, the incidence of ovarian cancer ranks third, and because many cases are already advanced when they are first diagnosed, its mortality rate ranks first. Surgery and postoperative chemotherapy are the main treatment methods for ovarian cancer. Compared with surgery alone, postoperative chemotherapy for ovarian cancer can significantly prolong the survival of patients, so the value of chemotherapy in the treatment of ovarian cancer has been fully affirmed. Because ovarian cancer is resistant to chemotherapeut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/09
Inventor 陈新莲王和刘珊玲
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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