Andrographolide Inhibits Proliferation Through Induces Apoptotic And Blocking The Cell Cycle In Bladder Cancer Cells

Objective:To explore the inhibitory effect of andrographolide on T24 and 5637 human bladder cancer cells in vitro and its related mechanisms,and provide a theoretical basis for the development of andrographolide as a potential therapeutic agent for clinical application.Methods:Treating T24 and 5637 bladder cancer cells with different concentrations of andrographolide,and using Cell Counting 8(CCK-8)kit to detect the inhibitory effect of andrographolide on proliferation of T24 and 5637 cells;after incubated with andrographolide for 24 H,the morphological changes of apoptotic tumor cells were observed by ordinary light microscope;The cell clony formation experiment was performed by observing the number of bladder tumor cell formation communities under different andrographolide concentrations;Flow cytometry was implemented to detect the apoptosis of T24 and 5637 cells under different concentrations of andrographolide,and analyze the effect of andrographolide on cell cycle;The effect of andrographolide on the levels of the P53,Caspase3,PARP1,Caspase9 apoptotic proteins in T24 and 5637 cells was examined by Western blotting.Results:Andrographolide can effectively inhibit the viability and proliferation of T24 and 5637 bladder cancer cells.There is a dose-effect relationship between the two,and the cell proliferation rate decreases with the increase of drug concentration;after 24 hours of treatment with andrographolide on tumor cells,it was observed by ordinary light microscopy that the cells in the control group grew well,the cell bodies were full,the cytoplasm was rich,and the nucleus was intact,while the drug-added group showed poor cell adhesion and increased dead cells.The viable cells showed obvious cell morphological changes;with the increase of the concentration of andrographolide,the proliferation and clony formation ability of T24 and 5637 bladder cancer cells decreased significantly;the results of flow cytometry showed that T24 and 5637 cells showed an increase in the proportion of early apoptotic cells as the concentration of andrographolide increased compared with the control group.The apoptosis rate was significantly higher than that of the control group.The apoptosis rate of tumor cells was statistically different(P<0.05);P53 protein expression levels were enhanced along with the increasing concentrations of andrographolide,protein expression levels of cleaved-PARP1 were un-regulated,moreover,proteins expression levels of Caspase3 and Caspase9 were increased.Conclusions:Andrographolide can inhibit the proliferation of bladder cancer T24 and 5637 cells in a certain concentration range,and has obvious dose-effect relationship;its inhibition on bladder cancer cells may be through inducing cell cycle arrest and activation of endogenous mitochondrial apoptosis pathway to achieve.