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Breast cancer multidrug-resistant cell strain constructed by virtue of epirubicin induction as well as construction method and application thereof

A multi-drug resistance, epirubicin technology, applied in tumor/cancer cells, microorganism-based methods, animal cells, etc. solution, etc.

Inactive Publication Date: 2014-11-12
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

2. It has inhibitory effect on the activity of topoisomerase Ⅱ (topoisomerase type Ⅱ), which leads to the disintegration of genomic DNA
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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  • Breast cancer multidrug-resistant cell strain constructed by virtue of epirubicin induction as well as construction method and application thereof
  • Breast cancer multidrug-resistant cell strain constructed by virtue of epirubicin induction as well as construction method and application thereof
  • Breast cancer multidrug-resistant cell strain constructed by virtue of epirubicin induction as well as construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Induction and establishment of human breast cancer multidrug-resistant cell line MDA-MB-231 / EPI

[0045] The invention adopts the in vitro induction method of gradually increasing the epirubicin concentration and intermittent action to establish the human breast cancer multidrug-resistant cell line MDA-MB-231 / EPI. Specific steps are as follows:

[0046] 1. Human breast cancer MDA-MB-231 cell line was incubated with DMEM medium (containing 10% fetal bovine serum, penicillin 100 U / ml, streptomycin 100 μg / ml) at 37°C, 5% CO 2 , Cultivated under 95% humidity conditions;

[0047] 2. When cultured to 70%-90% adherence rate, discard the supernatant, add epirubicin to make the effect concentration 0.05-0.07mg / l, at 37℃, 5% C0 2 , Continue culturing under 95% humidity conditions, replace with fresh culture medium after 24 hours, and continue culturing. After about 6-8 weeks, the cells grow stably and are passaged twice;

[0048] 3. Appropriately increase the conce...

Embodiment 2

[0052] Embodiment 2 Carry out morphological observation and biological characteristic identification to the MDA-MB-231 / EPI cell strain that the method of the present invention is established:

[0053] 1. Morphological observation

[0054] 1.1 Observe the morphology of living cells with an inverted phase-contrast microscope: take a bottle of MDA-MB-231 and MDA-MB-231 / EPI 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 MDA-MB-231 cells are polygonal, the shape is relatively consistent, and the intracellular structure is uniform (such as figure 1 shown); the size and shape of MDA-MB-231 / EPI cells changed, the cells were more slender, and there were more dark substances in the cells, especially near the cell membrane (as shown in Figure 2).

[0055] 1.2 Observation of cell morphology by HE staining method: After the cells were co...

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Abstract

The invention provides a breast cancer multidrug-resistant cell strain constructed by virtue of epirubicin induction, wherein the breast cancer multidrug-resistant cell strain has a collection number of CCTCC C201439. The invention further provides a method for constructing the breast cancer multidrug-resistant cell strain. According to the invention, an MDA-MB-231 / EPI drug-resistant strain is successfully established, wherein a drug-resistant index is 26.27 times, and obvious cross resistance is produced for taxol, etoposide and cis-platinum. A drug-resistant tumor cell model is provided for the following related researches so as to research drug-resistant tumor cell morphology and biological characteristics, research a tumor multidrug resistance mechanism, analyze sensitivity to chemotherapeutic drugs as well as screen chemotherapeutic drugs, develop tumor drug resistant reverse drugs and research and develop a more effective tumor therapeutic method, and the like.

Description

technical field [0001] The invention relates to a breast cancer multidrug-resistant cell line induced and established by epirubicin. Background technique [0002] Breast cancer is one of the most common malignant tumors in women. According to statistics, there are about 1.2 million cases of breast cancer in the world every year, and about 500,000 people die of breast cancer. Breast cancer is a systemic disease, and comprehensive treatment is very critical, among which chemotherapy is one of the important means that cannot be replaced. However, multidrug resistance often leads to treatment failure and late tumor recurrence and metastasis, seriously threatening the survival of patients. The drug resistance of tumor cells during chemotherapy often manifests as multidrug resistance, that is, the phenomenon of cross-resistance to multiple chemotherapeutic drugs with different structures and mechanisms of action at the same time, which is the main reason for the failure of chemot...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C12R1/91
Inventor 李敏张丽菡田莉张春雨
Owner LANZHOU UNIVERSITY
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