摘要
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 effects of berbamine on cell proliferation and apoptosis of PC-3 in vitro and the possible mechanisms involved. Methods The A values of PC-3 in each group with various concentrations （ 10,20,30,40 mg/L） of berbamine at various time intervals （ 24,48,72 h ） were detected with the MTT. The apoptosis rate and cell cycle of the PC-3 cells treated with berbamine were assessed by flow cytometry analysis. Transmission electron microscope was employed for the cell apoptosis. Immunohistochemical technique was used to determine the expression of bax, bcl-2 and survivin proteins after treatment with berbamine. Results After PC-3 cells were treated with 10,20,30 and 40 mg/L berbamine for 48 h, the inhibition ratio was （ 13.60 ± 1.49 ） %, （ 31.79 ± 2.31 ） %, （ 54.16 ± 3.09） % and （ 63.72 ± 2.46 ） % respectively （ compared with the controls, P 〈 0.05 ）. Cell proliferation of prostate cancer cell PC-3 was markedly inhibited by berbamine in a time-and-dose dependent manner. Af- ter PC-3 cells were incubated with 40 mg/L berbamine for 24,48 and 72 h,the cell apoptotic rate was （31.44 ±3.27）% ,（50.32 ±4.03）% and （63.46 ±3.75）% respectively （compared with the controls, P 〈0.05）. With the increase of berbamine concentration,the ratio of G0/G1 and S cells was increased and the ratio of G2/M was decreased. Under the transmission electron microscopy ,the apoptosis body was seen. Immunohistochemical technique showed the expression of bax in the PC-3 cells treated with 40 mg/L of berbamine was significantly increased and survivin significandy decreased （ P 〈 0.05 ）. There was no significant changes in the expression of bcl-2 （ P 〉 0.05 ）. Conclusion Berbamine can inhibit the proliferation of PC-3 cells by inducing apoptosis and affecting cell cycle in vitro in a time-and-dose-dependent manner. Berbamine induces apoptosis by down-regulating the expression of survivin protein and up-regulating the expression of bax protein.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第8期957-959,共3页
Chinese Journal of Experimental Surgery
关键词
小檗胺
前列腺癌
脱噬作用
Berbamine
Prostatic carcinoma
Apoptosis
