Human ovarian cancer multidrug resistant cell line
A technology for multidrug resistance and ovarian cancer, applied in the field of cell engineering
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Embodiment 1
[0020] Example 1 Induction and establishment of human ovarian cancer multidrug-resistant cell line HO-8910 / VP16
[0021] The human ovarian cancer multidrug-resistant cell line HO-8910 / VP16 was established by gradually increasing the concentration of VP16 and intermittently induced in vitro. The specific steps are as follows:
[0022] (1) Culture medium for human ovarian cancer cell line HO-8910 (RPMI-1640 medium containing 10% calf serum), 5% CO2, 37°C, culture to 70%-90% adherence rate, Remove the supernatant, add VP161000ng / ml impact culture for 1 hour, remove VP16100ng / ml culture solution, wash the blank RPMI-1640 culture solution twice, replace the proliferation medium and culture for 72-96h to expand the surviving cells. When the surviving cells expanded to 70%-90% adherence rate, repeat the shock culture with VP16100ng / ml culture solution for 1 hour and 9 times, that is, complete 10 shock cultures with fluorouracil and 1000ng / ml paclitaxel culture solution for 1 hour, an...
Embodiment 2
[0025] Morphological observation of embodiment 2 drug-resistant strains
[0026] The multidrug-resistant human ovarian cancer cell line HO-8910 / VP16 in the logarithmic growth phase was taken, and the cell morphology was observed under an inverted microscope. The results were as follows: Figure 6 Shown: the cell shape becomes larger, the nucleus shrinks, and the cells are easy to accumulate into clusters.
Embodiment 3
[0027] Example 3 Cell Growth Curve Determination
[0028] Cells were digested, counted, and prepared into a cell suspension with a concentration of 5×103 / mL, and 100 μL of cell suspension was added to each well of a 96-well cell culture plate (500 cells per well); the 6-well cell culture plate was placed at 37°C , cultured in a 5% CO2 incubator for 2d, 4d, 6d, 8d, and 10d respectively; stain the 96-well plate with MTT, λ=490nm, and measure the OD value; add 20μL MTT (5mg / mL) to each well, and continue in the incubator Incubate for 4 hours; discard the medium, add 150 μL DMSO to each well to dissolve, shake gently for 10 minutes; λ=490nm, read the OD value of each well with a microplate reader, take time as the abscissa, and OD value as the ordinate Coordinates to plot cell growth curves. For the cell growth curves of HO-8910 and HO-8910 / VP16, see figure 1 .
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