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目的探讨苦参碱抑制雄激素非依赖性前列腺癌细胞株PC-3增殖、诱导细胞凋亡的可能机制,为临床雄激素非依赖性前列腺癌的治疗提供一定的实验基础和理论依据。方法应用活性Caspase-3检测、Western blot印迹分析不同浓度苦参碱作用PC-3细胞后其Caspase-3表达的差异,并通过荧光免疫细胞化学染色、Western blot印迹分析研究不同浓度苦参碱对PC-3细胞Bax/Bcl-2表达的影响。结果Caspase-3检测显示不同浓度苦参碱均可上调Caspase-3表达。随着苦参碱浓度的增加,Caspase-3表达的荧光强度升高,Caspase-3/β-actin积分密度值亦升高,呈现剂量依赖性关系(P<0.01)。Bax、Bcl-2蛋白表达分析显示不同浓度苦参碱作用PC-3细胞后, Bax蛋白的表达增加,Bcl-2蛋白表达下降。Bax表达的荧光强度及Bax/β-actin积分密度值随着苦参碱浓度的增加而升高,呈现剂量依赖性关系(P<0.01);Bcl-2表达的荧光强度及Bcl-2/β-actin积分密度值随着苦参碱浓度的增加而下降,亦呈现剂量依赖性关系(P<0.01)。结论苦参碱抑制PC-3细胞增殖并诱导细胞凋亡可能涉及到了线粒体-细胞色素C/Caspase-9/Caspase-3途径。
Objective To investigate the possible mechanism of matrine in inhibiting proliferation and inducing apoptosis of androgen-independent prostate cancer cell line PC-3 and to provide some experimental and theoretical evidences for the treatment of androgen-independent prostate cancer. Methods The expression of Caspase-3 in PC-3 cells treated with different concentrations of matrine was detected by Western blot analysis with active Caspase-3 assay. Fluorescent immunocytochemistry and Western blot analysis were used to study the effects of different concentrations of matrine PC-3 cells Bax / Bcl-2 expression. Results Caspase-3 test showed that different concentrations of matrine can up-regulate Caspase-3 expression. With the increase of matrine concentration, the fluorescence intensity of Caspase-3 expression increased, and the integral density value of Caspase-3 / β-actin also increased in a dose-dependent manner (P <0.01). Bax and Bcl-2 protein expression analysis showed that the expression of Bax protein and Bcl-2 protein decreased after treated with different concentrations of matrine for PC-3 cells. The fluorescence intensity of Bax and the integral density of Bax / β-actin increased with the increase of matrine concentration in a dose-dependent manner (P <0.01). The fluorescence intensity of Bcl-2 and Bcl-2 / β The -actin integral density value decreased with the increase of matrine concentration, and also showed a dose-dependent relationship (P <0.01). Conclusion Matrine inhibits the proliferation of PC-3 cells and induces apoptosis possibly involving the mitochondrial-cytochrome C / Caspase-9 / Caspase-3 pathway.