Application of ursolic acid in preparation of anti-colon cancer drugs
A colon cancer and ursolic acid technology, applied in the field of anti-tumor drugs, can solve the problem that the survival rate of colorectal cancer patients has not been significantly improved, and achieve the effect of promoting the apoptosis of colon cancer cells
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Embodiment 1
[0025] Example 1 : Ursolic acid inhibits the proliferation of colon cancer cells
[0026] Human colon cancer HCT116, HCT8, LoVo, SW620, RKO and other cells in the logarithmic growth phase were selected, digested with 0.25% trypsin containing 0.1% EDTA, and collected the cells. Count manually using a cell counting plate and adjust the cell density to 1×10 4 Cells / well / 100 μL were inoculated in a 96-well culture plate and sealed with D-Hank's solution. Placed at 37, ℃ 5% CO 2 Routinely grow overnight in an incubator. After the cells adhered to the wall on the second day, the supernatant was carefully discarded. Except for the blank control group (using complete medium), each group was added with different concentrations of drugs (the final concentration of ursolic acid single drug group was 5 μM, 10 μM, 15 μM , 20 μM, 25 μM, 30 μM, 40 μM, 60 μM, 80 μM, 100 μM), each group was set up with 6 replicate wells, and D-Hank's solution was used to seal the edges around them. Place...
Embodiment 2
[0030] Example 2 : Ursolic acid inhibits colon cancer cell migration
[0031] Colon cancer cells in the logarithmic phase of growth were selected and counted after routine trypsin digestion and resuspension; the cell density was adjusted to 4×10 using complete medium containing fetal bovine and double antibodies 5 1 cell / well (24-well plate), add 1.5mL cell suspension to each well; let stand in the ultra-clean workbench for 15-30min, avoid shaking the cell culture plate; place at 37, ℃ 5% CO 2 Routinely cultivate overnight in the incubator until the cells adhere to the wall; the next morning, use a 200 μL imported yellow pipette tip with a ruler to complete the scratch in the ultra-clean bench, rinse gently with PBS twice, and add bearberry prepared by serum-free medium Acid drugs, complete the 0h photo under the electron microscope at 100 times; continue to 37, ℃ 5% CO 2 Routinely cultivated in the incubator for 48 hours, and completed 48 hours of photographing; the differ...
Embodiment 3
[0033] Example 3 : Ursolic acid inhibits autophagy and promotes apoptosis in colon cancer cells by down-regulating myeloid differentiation factor 88 in vitro
[0034] Human colon cancer HCT116 and RKO cells in the logarithmic growth phase were selected, and the cells were collected after routine trypsin digestion, and the cell density was adjusted to 25×10 4 Cells / well / 3mL were seeded in 6-well culture plate, 37, 5% CO 2 Incubate overnight in an incubator until the cells adhere to the wall. The next morning, set up control wells without any treatment, discard the complete medium in starvation-induced autophagy wells, replace with D-Hank's solution, and return to 37, 5% CO 2Continue culturing in the incubator for 1, 2, and 4 hours, collect the cells, extract total RNA and / or total protein, and perform q-PCR and / or Western blot detection of autophagy-related genes to verify the conditions for establishing the autophagy model.
[0035] On the basis of the colon cancer cell st...
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