Cervical cancer multidrug resistance cell strain established by cisplatin 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
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  • 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 dru...

Method used

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  • Cervical cancer multidrug resistance cell strain established by cisplatin induction
  • Cervical cancer multidrug resistance cell strain established by cisplatin induction
  • Cervical cancer multidrug resistance cell strain established by cisplatin induction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] SiHa / cDDP 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 the SiHa cells in the logarithmic growth phase, change the fresh culture medium, add cDDP, make the effect concentration be 0.1 μ g / 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) Properly increase the concentration of cDDP, at 37 ° C, 5% CO 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 i...

Embodiment 2

[0012] Morphological observation and identification of biological characteristics of the established SiHa / cDDP cell line:

[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 / cDDP cells in the logarithmic growth phase, observe the morphology of living cells under an inverted phase-contrast microscope after changing the medium, and take pictures. It can be seen that SiHa cells are spindle-shaped, with regular size and shape (as shown in Figure 1); SiHa / cDDP cells appear more slender, sometimes even linear (as shown in Figure 2).

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

[0017] Take SiHa and SiHa / cDDP 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 cover glass, put CO 2 Routine culture in an incubator. After the cel...

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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 cis-platinum (cDDP) is gradually increased from 0.1 mu g/ml to 0.2 mu g/ml; and drug resistant SiHa/ cDDP cell strain is established. The SiHa/cDDP cell can stably grow, transfer and reanimate in 2 mu g/ml cDDP, makes the cDDP generate drug resistant index (RI) of 16.131, and performs intersect drug on etoposide, taxol, vincristine, methotrexate, doxorubicin and mitomycin C except the cDDP. 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 multidrug-resistant cervical cancer cell line SiHa / cDDP by gradually increasing the concentration of the chemotherapeutic drug cisplatin (cDDP). 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 dif...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/09
Inventor 王和刘珊玲陈新莲
Owner THE WEST CHINA SECOND UNIV HOSPITAL OF SICHUAN
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