PI3K/AKT/Bcl-2信号通路介导人前列腺癌PC-3细胞凋亡及苦参碱的干预效果分析

PI3K/AKT/Bcl-2 signaling pathway mediates human prostate cancer PC-3 cell apoptosis and analysis on matrine intervention effect

目的探讨苦参碱(Mat)诱导人前列腺癌PC-3细胞凋亡的机制及其对磷脂酸肌醇3-激酶/丝氨酸苏氨酸激酶/B细胞淋巴瘤-2蛋白(PI3K/AKT/Bcl-2)信号通路的影响。方法采用不同质量-体积浓度(0.1、0.2、0.3、0.4、0.5 g/L)Mat处理PC-3细胞24、48、72 h,通过细胞增殖抑制试验检测PC-3细胞的增殖情况;采用流式细胞仪分析Mat对PC-3细胞周期分布的影响及凋亡率,运用蛋白质印迹法(Western blotting)检测用药前后细胞中PI3K、AKT、pAKT、Bax和Bcl-2蛋白的表达水平。结果经同一浓度Mat作用,48、72 h PC-3细胞增殖抑制率与24 h比较,差异均有统计学意义(P<0.05);处理48 h后,S期、G_0/G_1期0.2~0.5 g/L Mat组与对照组和0.1 g/L Mat组比较,差异均有统计学意义(P<0.05)。0.2~0.5 g/L Mat组细胞凋亡率明显高于对照组和0.1 g/L Mat组,差异均有统计学意义(P<0.05)。经0.1、0.2、0.3、0.4、0.5 g/L Mat处理48 h后,PC-3细胞PI3K、AKT、p-AKT、Bcl-2蛋白表达降低,Bax蛋白表达升高,差异均有统计学意义(P<0.05)。结论一定质量浓度范围Mat(0.2~0.5 g/L)能够抑制人前列腺癌PC-3细胞的增殖,Mat通过影响PI3K/AKT/Bcl-2信号通路诱导人前列腺癌PC-3细胞凋亡。

Objective To investigate the mechanism of matrine（Mat） inducing human prostate cancer PC-3 cells apoptosis and its effect on PI3K/AKT/Bcl-2 signaling pathway. Methods PC-3 cells were processed by different concentration of Mat（0.1,0.2,0.3,0.4,0.5 g/L） for 24,48,72 h. The PC-3 cell proliferation was detected by methyl thiazolyl tetrazolium.The effect of Mat on PC-3 cells cycle distribution and apoptosis rate were detected by flow cytometry. The expression levels of PI3 K,AKT,p-AKT,Bax and Bcl-2 protein before and after medication were analyzed by Western blot. Results By the action with the same concentration of Mat,the PC-3 cellular apoptosis rate had statistical difference between 48,72 h with 24 h（P〈0.05）;after 48 h of Mat treatment,the difference between the 0.2-0.5 g/L Mat group with control group and 0.1g/L Mat group at the stage S and G_0/G_1 had statistical significance（P〈0.05）.The apoptosis rate in the 0.2~0.5 g/L Mat group was significantly higher than that in the control group and 0.1 g/L Mat group,the differences were statistically significant（P〈0.05）.After 48 h of 0.1, 0.2, 0.3, 0.4, 0.5 g/L Mat treatment,the expression of PI3 K,AKT,p-AKT and Bcl-2 protein in PC-3 cells were decreased,while Bax protein expression was increased,the differences were statistically significant（P〈0.05）. Conclusion In a certain mass-volume concentration range（0.2 ~0.5 g/L）,Mat can inhibit human prostate cancer PC-3 cells proliferation,Mat induces human prostate cancer PC-3 cells apoptosis by influencing PI3K/AKT/Bcl-2 signal pathway.