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目的探讨小檗胺体外诱导雄激素非依赖性前列腺癌PC-3细胞凋亡的作用及其机制。方法噻唑蓝(MTT)法检测10、20、30和40 mg/L小檗胺作用于PC-3细胞24、48、72h的吸光度(A)值;流式细胞仪检测小檗胺作用后细胞凋亡率及细胞周期的变化;透射电镜观察细胞凋亡形态;免疫细胞化学法检测小檗胺作用后bcl-2、bax和Survivin表达变化。结果各浓度组小檗胺作用48h,细胞抑制率分别为(13.60±1.49)%、(31.79±2.31)%、(54.16±3.09)%和(63.72±2.46)%,与对照组比较差异均有统计学意义(P<0.05)。小檗胺对PC-3细胞的抑制作用呈时间-浓度依赖性;40 mg/L小檗胺作用24、48、72 h,细胞凋亡率分别为(31.44±3.27)%、(50.32±4.03)%和(63.46±3.75)%,与对照组比较差异有统计学意义(P<0.05)。随小檗胺浓度的增高,凋亡率随之升高,细胞周期呈现G_0/G_1期、S期细胞比例增多,G_2/M期细胞比例减少趋势;电镜扫描可见典型的“凋亡小体”;40 mg/L小檗胺作用72 h,PC-3细胞中bax的表达增加、Survivin表达减少(P<0.05),而bcl-2的表达差异无统计学意义(P>0.05)。结论小檗胺通过诱导细胞凋亡和影响细胞周期的方式抑制PC-3细胞的增殖,并具有时间和浓度依赖性;诱导凋亡的机制可能是通过下调Survivin、上调bax蛋白的表达来实现。
Objective To investigate the effect and mechanism of berbamine inducing apoptosis of androgen-independent prostate cancer PC-3 cells in vitro. Methods The absorbance (A) value of PC12 cells treated with 10, 20, 30 and 40 mg / L berbamine was detected by MTT method. The cell viability was detected by flow cytometry Apoptotic rate and cell cycle. The morphology of apoptotic cells was observed by transmission electron microscopy. The expressions of bcl-2, bax and Survivin were detected by immunocytochemistry after berbamine treatment. Results The inhibitory rates of berbamine in each concentration group for 48h were (13.60 ± 1.49)%, (31.79 ± 2.31)%, (54.16 ± 3.09)% and (63.72 ± 2.46)%, respectively, Statistical significance (P <0.05). The inhibitory effect of berbamine on PC-3 cells was time-dependent. The apoptosis rates of berbamine at (31.44 ± 3.27)% and (40.32 ± 4.03) )% And (63.46 ± 3.75)%, respectively, with statistical significance (P <0.05). With the increase of the concentration of berbamine, the apoptosis rate increased, the cell cycle showed G_0 / G_1 phase, the proportion of S phase cells increased, the proportion of G_2 / M phase cells decreased; electron microscopy showed a typical "apoptotic body The expression of bax in PC-3 cells was increased and the expression of Survivin was decreased (P <0.05), while the expression of bcl-2 was not statistically significant (P> 0.05). Conclusion Berbamine inhibits the proliferation of PC-3 cells by inducing apoptosis and affecting the cell cycle in a time-and-concentration-dependent manner. The mechanism of inducing apoptosis may be through down-regulating Survivin and up-regulating the expression of bax protein.