Study Of Effects And Mechanisms Of Evodiamine On Prolifertion And Apoptosis In Human Prostate Cancer DU145Cells In Vitro

Objective: To investigate evodiamine in vitro human prostate cancerDU145cells proliferation and apoptosis, and explore its possible mechanism.Methods:Human prostate cancer DU145cells were cultivated in vitro and thentreated with various concentrations of evodiamine. The effects of theproliferation was investigated by MTT assay. Apoptosis rate, cell cycledistribution were detected by Flow cytometry （FCM）. Using microscopeobserved the change of morphologic of DU145cells. Immunocytochemicalstaining method was used to detect the expression of PCNA in human prostatecancer DU145cells. Western blot analysis the expression of t-ERK p-ERKand Cleaved Caspase-3.The results were analyzed by SPSS13.0, and weestablished the standard of statistic significance as P＜0.05.Results:1Evodiamine can inhibit the proliferation of DU145cells: MTT assaysresults suggested that evodiamine inhibits DU145proliferation （P<0.05）.Western blot analysis and Immunocytochemistry staining suggested thatevodiamine down regulated PCNA and p-ERK. It is no significant changes inexpression level of t-ERK.2Evodiamine on cell cycle of DU145: After DU145cells treated withvarious doses of evodiamine (10^-7M、10^-6M、10^-5M) for24hours, evodiamine(10^-6M、10^-5M) caused the accumulation of cell cycle at G₂/M phase inDU145cells. The increase in G₂/M population by EVO was accompanied by asignificant decrease in both G1and S phase.3Evodiamine can induce apoptosis in DU145cells: Apoptosis cellsincreased significantly after DU145cells treated with evodiamine (10^-7M、10^-6 M、10-5M) for72hours. This phenomenon showed concentration dependent.By one-way ANOVA, Apoptosis percentage （AP） were significantly differentamong concentration groups （P<0.05）. Western blot analysis suggested thatafter treated with various doses of evodiamine for24hours, the correspondentexpression of Cleaved Caspase-3were significantly increased in DU145cells.The increasing expression of Cleaved-caspase3existed significant differenceamong concentration groups （P<0.05）.Conclusions:1Evodiamine inhibits the proliferation of human prostate cancer DU145cells, this phenomenon may be achieved by inhibiting the phosphorylationlevel of ERK.2Evodiamine can induce apoptosis on DU145cells. The activation ofCaspase-3and up-regulation of Cleaved Caspase-3is one of the mechanismsof DU145cell apoptosis induced by evodiamine can be speculated.