Application of thiazole-pyrazoline derivatives in preparation of medicine for resisting non-small cell lung cancer
A non-small cell lung cancer, thiazole technology, applied in the application field of thiazole-pyrazoline derivatives in the preparation of anti-non-small cell lung cancer drugs, to achieve the effect of inhibiting the growth of A549 cancer cells and promoting the autophagy of A549 cells
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Embodiment 1
[0041] Example 1: Detection of survival rate by SRB method after CPT treatment of A549 cells.
[0042] (1) Add different concentrations of the compound CPT to treat the cells pre-planted in 96-well plates. (2) After compound treatment for 24 hours and 48 hours, the old culture medium was discarded, 100 μL of trichloroacetic acid was added, and the cells were fixed at 4° C. for 1 hour. (3) Discard the fixative. Wash five times with double-distilled water in gentle motions, and dry at room temperature. (4) Add 50 μL of SRB to each well, shake at room temperature for 10 minutes. (5) Wash 5 times with 1% acetic acid and dry at room temperature. (6) Add 100 μL of 10 mM buffer solution Tris base to each well and place on a shaker for 10 minutes. (7) Select 540nm excitation light to measure OD value.
[0043] The results show that: 5μM, 10μM CPT treatment of A549 cells for 24h and 48h can significantly inhibit the survival of A549 cells, see figure 1 .
Embodiment 2
[0044]Example 2: After A549 cells were treated with CPT, changes in cell morphology and number were detected under an inverted microscope.
[0045] A549 cells were seeded in a 6 cm-diameter petri dish at 37°C in CO 2 After culturing in the incubator for 12 hours, different concentrations of CPT were added for 12 hours, 24 hours and 48 hours, and then placed under an inverted microscope to photograph the cell morphology and observe the number of cells.
[0046] The results showed that: under the above conditions, 5μM and 10μM CPT could change the cell morphology, and some cells showed changes such as narrow, narrow, elongated and rounded, which significantly reduced the number of A549 cells. See figure 2 .
Embodiment 3
[0047] Example 3: After A549 cells were treated with different concentrations of CPT, the level of autophagy was detected by western blot.
[0048] A549 cells were seeded in a 6 cm-diameter petri dish at 37°C in CO 2 After culturing in the incubator for 12 hours, add different concentrations of CPT to treat for 24 hours, lyse the cells, collect the cell lysate, centrifuge at 12000g for 15 minutes at 4°C, take the supernatant, and use western blot to detect LC3B-I and LC3B-II in the cells treated in different ways protein level.
[0049] The results showed that: A549 cells were treated with 1 μM, 5 μM, and 10 μM CPT for 24 hours, which significantly promoted the increase of LC3B-Ⅱ levels and promoted the autophagy of A549 cells. See image 3 .
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