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Cervical cancer multidrug resistance cell strain established by etoposide induction

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

Inactive Publication Date: 2009-08-12
THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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  • Summary
  • 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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  • Cervical cancer multidrug resistance cell strain established by etoposide induction
  • Cervical cancer multidrug resistance cell strain established by etoposide induction
  • Cervical cancer multidrug resistance cell strain established by etoposide induction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] SiHa / VP16 multi-drug resistant cell line is established according to the following steps: (1) Human cervical cancer SiHa cell line, after recovery, use DMEM culture medium (containing 10% calf serum, penicillin 100U / ml, streptomycin 100ug / ml) At 37°C, 5% CO 2 , 95% humidity conditions; (2) take SiHa cells in the logarithmic growth phase, change fresh culture medium, add VP16, make the effect concentration be 0.1μg / ml, in 37 ℃, 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 2 μg / ml VP16 are obt...

Embodiment 2

[0012] The SiHa / 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 SiHa and SiHa / VP16 cells in the logarithmic growth phase, and observe the morphology of living cells under an inverted phase-contrast microscope after changing the medium and take pictures. It can be seen that the three-dimensional sense of SiHa / VP16 cells is enhanced, sometimes showing a short and fat spindle shape, but generally speaking, it is not much different from SiHa cells. (As shown in Figure 1 and 2)

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

[0017] Take SiHa and SiHa / VP16 cells in the logarithmic growth phase, digest with 0.25% trypsin to make a single cell suspension, and adjust the density to about 1×10 5 / ml, inoculated on a petri dish covered with a ...

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Abstract

The invention relates to a multidrug resistant cell strain for cervical carcinoma, which is characterized in that a human cervical carcinoma SiHa 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 4 mu g / ml; and drug resistant SiHa / VP16 cell strain is established. The SiHa / VP16 cell can stably grow, transfer and reanimate in 4 mu g / ml VP16, makes the VP16 generate drug resistant index (RI) of 24. 019, and performs intersect drug on MTX, cDDP, VCR, 5-Fu and DOX except 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 cell line of human cervical cancer. Specifically, it involves using human cervical cancer SiHa as the parent cell, and establishing the cervical cancer multi-drug resistant cell line SiHa / VP16 by gradually increasing the concentration of the chemotherapeutic drug etoposide (VP16). Background technique [0002] Cervical cancer seriously endangers women's health, and its incidence rate ranks second among female malignant tumors. There are about 500,000 new cases of cervical cancer in the world every year, and the number of new cases in my country accounts for about 1 / 3 of the total number of cases in the world. Chemotherapy is an important method for the treatment of cervical cancer, but a common and difficult problem in chemotherapy is the generation of multidrug resistance, that is, the phenomenon of cross-resistance of tumors to multiple chemotherapeutic drugs with di...

Claims

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Application Information

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