Effect Of Betulinic Acid And Dictamnine On Bone Metastasis Of Prostate Cancer

一、Effect of Betulinic Acid on bone metastasis of prostate cancer.ObjectiveTo evaluate the effect and mechanism of betulinic acid(BetA)on the proliferation,migration,invasion and epithelial-mesenchymal transformation(EMT)of PC-3 cells induced by aerobic glycolysis in prostate cancer(PCa).To explore the therapeutic effect and significance of betulinic acid on prostate cancer.MethodsCultivate human prostate cancer cells(PC-3cells),using PC-3 betulinic acid cells,through the comparison between the experimental group and control group of determination of cell survival and cell proliferation,glycolysis metabolism,cell migration and invasion test,cell sphere formation and colony formation test to understand betulinic acid effects on prostate cancer(PC-3)cells.Real-time PCR was used to obtain and measure the mRNA levels of several major rate-limiting enzymes in glycolysis,such as hexokinase 2(HK2)and phosphofructokinase 1(PFK1),and Westernblot was used to analyze and compare the levels of these target proteins between the experimental group and the control group.The expression levels of target proteins such as N-cadherin,fibronectin,vimentin and ZEB1 during EMT were determined by the same method.ResultsBetulinic acid can inhibit the survival and proliferation of PC-3 cells,and reduce the expression of cyclin A and cyclin-dependent kinase.Betulinic acid also decreased glucose uptake and consumption,intracellular lactic acid level,extracellular acidification rate,and the expression and activity of glycolytic rate-limiting enzymes hexokinase 2(HK2)and phosphofructokinase 1(PFK1),which are involved in hypoxia-inducible factor-1α(HIF-1α)and transcriptional coactivator associated protein(YAP).In addition,betulinic acid effectively inhibited tumor metastasis by inhibiting the migration,invasion,spherogenesis and colony formation of PCA cells in vitro.Betulinic acid inhibited the EMT process in PC-3cells,which was confirmed by the changes in the expression of EMT-related markers.The inhibitory effect of low dose BetA on EMT of PCA cells was dose-dependent.ConclusionBetulinic acid(BetA)can inhibit PCa cell proliferation,metastasis,and epithelium-mesenchymal transformation(EMT),and treatment with betulinic acid may be a potentially effective treatment option.This justifies the potential of betulinic acid as a practical chemoprophylaxis for prostate cancer patients.二、Effect of Dictamnine on bone metastasis of prostate cancer.Objective To evaluate the effect and mechanism of DIC on PC-3 cell proliferation,metastasis and epithelial-mesenchymal transformation.To explore the therapeutic effect and significance of dictamnine on prostate cancer.Methods Human prostate cancer cells(PC-3 cells)were cultured and PC-3 cells were treated with dictamnine.The effects of dictamnine on PC-3 cells were studied by comparing the cell proliferation,cell migration and invasion test and cell morphological changes between the experimental group and the control group.The target protein levels during the process of epithelial-mesenchymal transformation were measured by Westernblot.(e.g.Snail,E-Cadherin,β-Catenin,etc.)Results The ability of PC-3 cells proliferation,migration and invasion were all decreased after the treatment of dictamnine.The mRNA and protein expressions of E-cadherin in PC-3 cells were increased,and the mRNA and protein expressions of Vimentin,Snailin and β-catenin in the nucleus were decreased after the treatment of dictamnine(P < 0.01).In addition,the mRNA and protein expressions of Snail in PC-3 cells were increased and the mRNA and protein expressions of E-cadherin were decreased after the combined treatment with Licl(P < 0.01).Conclusion Dictamnine can inhibit the proliferation,migration and invasion of PC-3 cells,and inhibit the epithelial-mesenchymal transformation of PC-3 cells.The mechanism may be that dictamnine inhibits epithelial-mesenchymal transformation of PC-3 cells by inhibiting Wnt/β-catenin/Snail signaling pathway.