A breast cancer multidrug-resistant cell line induced by epirubicin and its construction method and application
A multi-drug resistance, epirubicin technology, applied in the direction of tumor/cancer cells, microorganism-based methods, animal cells, etc., can solve the disintegration of genomic DNA, easy to produce drug resistance, and affect the drug resistance of tumor cells. degree, etc.
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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 concent...
Embodiment 2
[0052] Embodiment 2 Carry out morphological observation and biological characteristic identification to the MDA-MB-231 / EPI cell strain that application method of the present invention establishes:
[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...
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