Study On The Mechanism Of Kaempferol Extracted From Zhuang Medicine Shannai On Prostate Cancer

Objective: In this study,we investigated the effects and mechanism of action of kaempferol on the proliferation and apoptosis of two different types of prostate cancer cells(androgen-dependent and androgen-independent).To comprehensively evaluate the anti-prostate cancer effect of kaempferol in vivo and in vitro,and to provide the scientific basis for the further development of kaempferol.Methods: 1.To investigate the effect of kaempferol on the cell viability of prostate cancer cells(PC-3,DU145,22Rv1,LNCa P)by MTT assay.2.Plate colony formation assay was used to examine the effect of kaempferol on the proliferation of PC-3,DU145,and 22Rv1 cells.3.Flow cytometry was used to test the effect of kaempferol on the apoptosis of PC-3,DU145,and22Rv1 cells.4.Using cell cycle kit to determine the effect of kaempferol on cell cycle of PC-3 and 22Rv1.5.Real-time fluorescence quantitative Polymerase Chain Reaction(q PCR)was used to detect the effects of kaempferol on the expression of proliferation marker Ki67 in prostate cancer cells(PC-3,22Rv1).6.Immunofluorescence and enzyme-linked immunosorbent assay(Elisa)were used to investigate the effect of kaempferol on the expression of Ki67 protein in prostate cancer cells(PC-3,22Rv1).7.The xenograft tumor model of zebrafish with prostate cancer was established by microinjection of 22Rv1 cells into the yolk sac of zebrafish.After continuous intervention with kaempferol for 6 days,the anti-prostate cancer activity of kaempferol in vivo was assessed by photographic method to calculate the fluorescence area.Results: 1.Kaempferol inhibited the cell viability of four prostate cancer cell lines(PC-3 cells,DU145 cells,22RV1 cells,LNCa P cells).2.The results of plate clone formation assay suggested that kaempferol could significantly inhibit the colony formation of prostate cancer cells PC-3,DU145,and 22Rv1 in at a dose-dependent manner.3.Flow cytometry showed that kaempferol could induce apoptosis of prostate cancer cells PC-3,DU145,and 22Rv1.4.The results of the cell cycle showed that kaempferol could block the PC-3 cell cycle at G2/M phase and the 22Rv1 cell cycle at G1 phase.5.Kaempferol could inhibit the expression of Ki67 m RNA in PC-3 cells and 22Rv1 cells.6.Kaempferol could up-regulate the expression of Ki67 protein in PC-3 cells and 22Rv1 cells.7.Kaempferol showed a good anti-tumor effect in the prostate cancer zebrafish xenograft tumor model.Six days after administration,the fluorescence area of the experimental group was smaller than that of the model group,and the difference was statistically significant(P< 0.05),suggesting that kaempferol can effectively inhibit the growth of tumors in zebrafish.Conclusion: 1.Kaempferol can inhibit the proliferation of prostate cancer cells by inducing apoptosis and blocking cell cycle of prostate cancer cells.2.Kaempferol have different effects on the cell cycle of different types of prostate cancer by regulating the expression of Ki67.3.Kaempferol can effectively inhibit the growth of tumors in zebrafish and has anti-prostate cancer effect in vivo.