Ovarian cancer multi-drug resistant cell line induced by triptolide and its application

A technology of triptolide and multidrug resistance, which is applied in the field of tumor biology and can solve problems affecting the degree of drug resistance of tumor cells

Active Publication Date: 2018-03-09
HARBIN MEDICAL UNIVERSITY
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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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  • Ovarian cancer multi-drug resistant cell line induced by triptolide and its application
  • Ovarian cancer multi-drug resistant cell line induced by triptolide and its application
  • Ovarian cancer multi-drug resistant cell line induced by triptolide and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Induction and establishment of ovarian cancer multidrug-resistant cell line SKOV3 / TPL

[0021] SKOV3 / TPL multidrug-resistant cell line was established according to the following steps: (1) Human ovarian cancer SKOV3 cell line was recovered with 1640 medium (containing 10% calf serum, penicillin 100U / ml, streptomycin 100U / ml) At 37℃, 5%CO 2 (2) Take logarithmic growth phase SKOV3 cells, change to fresh culture medium, add triptolide (TPL) to make its concentration 10nM, at 37℃, 5%CO 2 Continue to culture under 95% humidity conditions, change the medium at appropriate time, and maintain the drug concentration of 10 nM until the viable cells can grow and pass stably at this concentration; (3) Appropriately increase the concentration of TPL, at 37 °C, 5% CO 2 2. Continue culturing under 95% humidity conditions, change the medium at appropriate time, and maintain the drug concentration until the viable cells can grow and subculture stably at this concentration; (4...

Embodiment 2

[0024] Example 2 Morphological observation and identification of biological characteristics of drug-resistant cell line SKOV3 / TPL

[0025] 1.1 Morphological observation

[0026] (1) Inverted phase contrast microscope to observe the morphology of living cells

[0027] One bottle of SKOV3 and SKOV3 / TPL cells in the logarithmic growth phase was taken. After the medium was changed, the morphology of the living cells was observed and photographed under an inverted phase contrast microscope. It can be seen that the SKOV3 cells are polygonal, the shape is relatively consistent, and the intracellular structure is uniform ( figure 1 ); SKOV3 / TPL cells were larger, with different shapes, enlarged nuclei and cytoplasm, increased multinucleoli, and more dark substances aggregated in the cells, especially near the cell membrane ( figure 2 ).

[0028] (2) HE staining to observe cell morphology

[0029] Take logarithmic growth phase SKOV3 and SKOV3 / TPL cells, digest with 0.25% trypsin t...

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Abstract

The invention relates to a multidrug-resistant ovarian cancer cell line induced and established by triptolide and use thereof. The cell line is named SKOV3 / TPL, and the microorganism preservation number is CGMCC No. 10306. In the parental cells, the concentration of Triptolide (TPL), a monomeric extract of the traditional Chinese medicine Triptolide, was gradually increased from 10nM to 400nM, and the multidrug-resistant ovarian cancer cell line SKOV3 / TPL was established. The multidrug-resistant ovarian cancer cell line SKOV3 / TPL can stably grow, passage and recover in 400nM TPL, and its resistance index (RI) to TPL is 50.95. ‑FU, Ara‑C, GEM, VP‑16, MMC, ADM, MIT, TAX, VCR have cross-resistance. The establishment of the multidrug-resistant ovarian cancer cell line of the present invention provides a drug-resistant tumor cell model for tumor-related research, and the action mechanism of the drug can be studied by comparing its genetic difference with the parental cell.

Description

technical field [0001] The invention relates to a drug-resistant human ovarian cancer cell line and use thereof, in particular to the establishment of multi-drug resistance of ovarian cancer by gradually increasing the action concentration of a chemotherapeutic drug triptolide (TPL) by using human ovarian cancer SKOV3 as a parental cell. The cell line SKOV3 / TPL belongs to the field of tumor biology. Background technique [0002] In the latest worldwide statistics on the incidence of tumors, the incidence of ovarian cancer ranks third among the malignant tumors of female reproductive organs, and the mortality rate ranks first, which seriously endangers women's health and life. Chemotherapy is one of the important means of ovarian cancer treatment and plays an important role in the comprehensive treatment of ovarian cancer, but clinical chemotherapy resistance is the main reason for the failure of ovarian cancer chemotherapy. The development of drug resistance is multifactori...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12Q1/02C12R1/91
Inventor 张雅芳王庚王莉李娜吕晓红
Owner HARBIN MEDICAL UNIVERSITY
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