Effects Of Diterpene Alcohols On Kidney Cancer Renca Cell Cycle, Apoptosis And Migration And Invasion Ability

Objective:Based on the previous cell experiments in vitro: the Chinese medicine Semen Euphorbiae Lathyridis has a good pharmacodynamic effect on the inhibition of Renca cell proliferation.The effects of Semen Euphorbiae Lathyridis on the morphology,invasion and metastasis,cell cycle and apoptosis of Renca cell were studied by using modern molecular biology experimental technology.In order to provide experimental basis and theoretical basis for the clinical treatment of renal cancer research and development of new Chinese medicine treatment drugs.Method:1.Cell scratch test: the cells were inoculated into the 6-well plate.After the cells adhered to the wall,they were starved with serum-free medium for 6 hours,scratched with "1" shape with 200 ? l gun head perpendicular to the 6-well plate,and washed twice with PBS buffer.The best effective dose of dauricinol was 9000 ? g / ml in the medium without fetal bovine serum.The concentration of dauricinol was(9000 ? g / ml)in the experimental group,and the control group was added in the medium group.At the beginning of the scratch and 24 hours after the treatment,the width of the scratch was observed and photographed under the inverted microscope,and the effect of diterpenoid alcohol on Renca cells was recorded and photographed under the microscope.2.Transwell chamber method: transwwell chamber method was used to detect the number of cells passing through the membrane of Renca cell after being treated with chrysophane diterpenoid alcohol for 48 hours.Each experiment was repeated three times independently.The results were analyzed by mathematical statistical analysis.3.Flow cytometry(FCM): to detect the changes of proliferation cycle and apoptosis of Renca cell line induced by diterpenoid alcohol.After 24 hours treatment,the single cells were collected from the experimental group and the blank control group,and the apoptosis rate and proliferation cycle were detected by annexin V-FITC method and PI single staining method respectively.Flow job software was used to analyze the results.4.polyacrylamide gel electrophoresis results: compared with the control group,the expression of Pro apoptotic protein Bax and Caspase-3 increased significantly,and the expression of anti apoptotic protein Bcl-2 decreased.Conclusion:1.Chinese herbal medicine Semen Euphorbiae Lathyridis can inhibit the invasion and metastasis of Renca cells,and affect the cell morphology;2.The molecular mechanism of the inhibition of Renca cell proliferation by Semen Euphorbiae Lathyridis is probably related to the inhibition of cell division cycle and the induction of apoptosis;3.The molecular mechanism of apoptosis of Renca cells induced by Semen Euphorbiae Lathyridis is probably related to the expression of Bcl-2,Bax and caspase-3.