Method for establishing paclitaxel (PTX)-resistant ovarian cancer cell model

A technology of ovarian cancer cells and paclitaxel, applied in tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problem of low drug resistance index

Inactive Publication Date: 2015-03-04
JINSHAN HOSPITAL FUDAN UNIV
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Problems solved by technology

[0006] All of the above have established paclitaxel-resistant ovarian cancer cell models through the method of step-by-step induction, but the drug resistance index is still low. If the c

Method used

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Embodiment 1

[0026] 1. Materials

[0027] OVCAR-3 cells and SK-OV-3 cells were purchased from ATCC, USA.

[0028] Paclitaxel (PTX) was purchased from Taiji Group Sichuan Taiji Pharmaceutical Co., Ltd.: 30mg / 5ml / branch.

[0029] Paclitaxel working solution was prepared with cell culture medium (RPMI-1640+10% FBS, purchased from Gibco, USA).

[0030] 2. Steps of building a plant

[0031] 1) OVCAR-3 cells and SK-OV-3 cells were cultured with RPMI-1640+10% FBS+100 U / ml penicillin+0.1 mg / ml streptomycin.

[0032] 2) The two cell lines were intervened with PTX at an initial concentration of 0.01 μM, and the drug-free medium (RPMI-1640 + 10% FBS + 100 U / ml penicillin + 0.1 mg / ml streptomycin) was replaced after 24 hours of PTX administration.

[0033] 3) After the cells grow stably, the shape is plump, there are no floating dead cells, and the passage density is 30%, it can grow to 90% in 3 to 5 days, then start normal passage (P) and increase the concentration of PTX.

[0034] OVCAR-3 mother...

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Abstract

The invention relates to a method for establishing a paclitaxel (PTX)-resistant ovarian cancer cell model. The method comprises the following specific steps: intervening an OVCAR-3 cell and an SK-OV-3 cell by virtue of PTX, replacing a PTX-free culture medium, intervening the OVCAR-3 cell and the SK-OV-3 cell until growth of cells returns to be normal, namely, under the conditions that the cells stably grow, morphologies are full, no floating dead cells are produced and the passage density is 30%, enabling the cells to grow 90% in 3-5 days, starting to carry out normal passage and improving the concentration of PTX, wherein the intervention time for each time is 24 hours; and finally obtaining the PTX-resistant ovarian cancer cell model having the preservation numbers of CCTCC C2014203 and CCTCC C2014204. According to the method, by closely observing growth states of cells and selecting the intervention moment, the established cell model has higher level of drug-resistance coefficients and good resistance stability and can be used for the research on ovarian cancer drug-resistant mechanism, ovarian cancer drug-resistant reversion, prevention of chemotherapy resistance and the improvement on chemosensitivity.

Description

【Technical field】 [0001] The present invention relates to the establishment of a cell model, in particular to the establishment of a paclitaxel-resistant ovarian cancer cell model. 【Background technique】 [0002] Ovarian cancer is the first malignant tumor of the reproductive tract that causes female death. The treatment is mainly paclitaxel combined with platinum-based chemotherapy on the basis of tumor cytoreductive surgery. Chemotherapy works very poorly. Ordinary ovarian cancer cell lines have limitations in studying the drug resistance mechanism of ovarian cancer, while ovarian cancer drug resistant cell lines can better study the drug resistance mechanism from the perspective of molecular biology, which will help to further find the treatment of ovarian cancer after drug resistance method. [0003] Chinese journal "Tumor" 2006, No. 8, published a paper "Establishment of ovarian cancer paclitaxel-resistant cell line OC3 / TAX300 and gene expression profiling analysis", ...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C12R1/91
Inventor 许国雄张凌云
Owner JINSHAN HOSPITAL FUDAN UNIV
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