Establishment method and application of paraquat-resistant cell line of human lung cancer
A technology of paraquat and cell lines, applied in the field of cell engineering, to achieve the effect of improving the success rate of induction and obviously resisting the toxicity of PQ
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Embodiment 1
[0028] Example 1 Induction and establishment of paraquat-resistant cell lines of human lung cancer
[0029] The human lung cancer paraquat-resistant cell line A549 / PQ was established according to the following steps: (1) A549 cells were routinely cultured in 1640 medium containing 10% fetal bovine serum at 37°C and 5% CO 2 . Cells in the logarithmic growth phase were used for experiments each time. (2) Establish cell drug-resistant strains by using PQ induction method with increasing concentration and periodic exposure: the concentration of PQ in the medium first starts from 100 μmol / L, and after adding the drug for 24 hours, change the medium to remove the drug, and wait for the cells to re-grow to the logarithmic phase Afterwards, the same concentration of PQ was added again for stimulation, and this was repeated 3 times. Then extend the exposure time to 48h (100μmol / L exposure for 48h), change the medium to remove the drug, and after the cells re-grow to the logarithmic p...
Embodiment 2
[0031] Example 2 Morphological observation of human lung cancer paraquat-resistant cell line A549 / PQ and human lung cancer A549 cells
[0032] The human lung cancer cell lines A549 and A549 / PQ cells in the logarithmic growth phase were routinely digested and passaged, and the cell morphology was observed with a Leica optical microscope 24 hours after the cells were completely adhered to the wall, and the photos were taken with a Nikon camera. The results are as follows: figure 1 shown.
Embodiment 3
[0033] Example 3 Cell Counting Method Drawing Human Lung Cancer Paraquat-resistant Cell Line A549 / PQ and Human Lung Cancer Cell Line A549 Cell Growth Curve and Determination of Doubling Time
[0034] Take 50,000 parental A549 cells in mid-logarithmic growth and good growth conditions, and human lung cancer paraquat-resistant cell line A549 / PQ cells to make cell suspensions and inoculate them into culture flasks, 21 flasks of each cell. After 24 hours, the cell counting was started, and the counting was continued for seven days, with three flasks of cells per day, and the cell growth curve was drawn according to the counting results. Then calculate the doubling time (DT) according to the formula, DT=t×lg2 / (lgNt-lgN0), t is the cell culture time, N0 is the average number of cells measured after inoculation 24h, and Nt is the average number of cells after culturing th value, the result is as figure 2 and 3 shown.
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