PI3K/AKT信号通路阻断药联合小檗碱对前列腺癌DU145细胞生长的抑制作用

目的 探讨采用磷脂酰肌醇3-激酶(PI3K)抑制药——LY294002抑制磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/AKT)信号通路联合小檗碱对前列腺癌DU145细胞生长的抑制作用.方法 采用MTT法检测细胞增殖情况;采用流式细胞仪检测细胞凋亡率和细胞周期分布情况;采用Western blot检测细胞中PI3K、p-AKT、cyclin D1、bcl-2、BAX蛋白表达情况.结果 随着小檗碱作用浓度的增加,DU145细胞的增殖率逐渐降低;不同浓度小檗碱与LY294002联合处理对DU145细胞增殖的抑制作用均较相同浓度小檗碱单独处理增强(P﹤0.05).与空白对照组相比,小檗碱组DU145细胞的凋亡率增高(P﹤0.05),G0/G1期细胞所占比例增高(P﹤0.05),PI3K、p-AKT、cyclin D1、bcl-2蛋白表达水平均下调(P﹤0.05),BAX蛋白表达水平上调(P﹤0.05).与小檗碱组和空白对照组相比,小檗碱+LY294002组DU145细胞的凋亡率增高(P﹤0.05),G0/G1期细胞所占比例增高(P﹤0.05),PI3K、p-AKT、cyclin D1、bcl-2蛋白表达水平均下调(P﹤0.05),BAX蛋白表达水平上调(P﹤0.05).结论 小檗碱能够抑制DU145细胞增殖,并促进细胞凋亡和细胞周期阻滞,采用LY294002阻断PI3K/AKT信号通路能够明显增强小檗碱对前列腺癌DU145细胞生长的抑制作用.

Inhibitory effect of PI3K/AKT signaling pathway blocker combined with berberine on the growth of prostate cancer DU145 cells

Objective To investigate the inhibitory effect of PI3K/AKT signaling pathway blocker, LY294002, in combination with berberine on the growth of prostate cancer DU145 cells. Method MTT assay was utilized to detect the cell proliferation;flow cytometry was used to detect cell apoptosis and cell cycle distribution;the protein expression of PI3K, p-AKT, cyclin D1, bcl-2 and BAX in cells were detected by Western blot. Result As the concentration of berber-ine increased, the proliferation rate of DU145 cells was decreased; different concentration of berberine combined with LY294002 demonstrated enhanced inhibitory effect as compared with berberine treatment alone (P<0.05). Compared with the blank control group, DU145 cells treated with berberine showed higher apoptosis rate (P<0.05), higher proportion of cells in G0/G1 phase, but reduced expression of PI3K, p-AKT, cyclin D1, and bcl-2 protein (P<0.05), though increased BAX protein expression (P<0.05). Comparing with berberine treatment group and blank control group, cells treated with berberine+LY294002 had higher apoptosis rate (P<0.05), higher proportion of cells in G0/G1 phase (P<0.05), and the pro-tein expression of PI3K, p-AKT, cyclin D1 and bcl-2 were reduced (P<0.05), but the protein expression of BAX was up-regulated (P<0.05). Conclusion Berberine can suppress DU145 cell proliferation, promote cell apoptosis and cell cycle arrest, blocking PI3K/AKT signaling pathway with LY294002 can significantly enhance the inhibitory effect of berberine on the growth of prostate cancer DU145 cells.