喜树碱(CPT)诱导前列腺癌PC-3细胞凋亡及其机制的研究  

Effect of Camptothecin(CPT) on the proliferation and apoptosis of PC-3 cells

作　　者：赵行宇[1] 赵容[1] 田丰雨 侯建成[1] 张万生[1] 张巍[1] ZHAO Xing-yu;ZHAO Rong;TIAN Feng-yu;HOU Jian-cheng;ZHANG Wan-sheng;ZHANG Wei(Department of Biochemistry of Jilin Medical University,Jilin Jilin 132013,China)

机构地区：[1]吉林医药学院生物化学教研室,吉林吉林132013

出　　处：《毒理学杂志》2018年第4期264-267,共4页Journal of Toxicology

基　　金：国家自然科学基金(21102055);吉林省科技厅科技发展项目(20130101157JC);吉林省教育厅"十二五"科学技术研究项目(2012-334);吉林省教育厅"十二五"科学技术研究项目(2014-363);国家大学生创新创业项目(201713743003)

摘　　要：目的测定不同浓度的喜树碱(CPT)对前列腺癌PC-3细胞的促凋亡作用,并进一步探讨其促凋亡机制。方法人前列腺癌PC-3细胞,加入不同浓度的CPT(10~100μmol/L),继续培养24 h。采用形态学观察及MTT方法检测细胞增殖,流式细胞仪测细胞早期凋亡率。Western blot法检测凋亡相关蛋白Bcl-2、Bax、Cleaved Caspase-3和PARP的表达。结果形态学观察及MTT试验显示,不同浓度的CPT可导致PC-3细胞形态改变,并对其生长有明显抑制作用,具有剂量依赖性,其IC50为23.25μmol/L;细胞凋亡检测表明不同浓度的CPT可使早期凋亡比率增加。Western blot结果显示,不同浓度CPT可使PC-3细胞Bcl-2蛋白表达降低,而Bax和Cleaved Caspase-3蛋白,89-k Da PRPP蛋白表达增加且呈剂量依赖性。结论 CPT可以诱导PC-3细胞凋亡,其作用机制可能为激活Caspase依赖途径,并活化其作用底物PARP来实现。Objective To study the effect of different concentration of CPT on the apoptosis of human prostate cancer cells（ PC-3）,and to explore its mechanism. Methods The cultured PC-3 cells in vitro were divided into control group and CPT-treated groups（ 10 ~100 μmol/L）. The viability of PC-3 cells were detected by MTT assay. The morphology changes of PC-3 cells was observed with inverted microscope. The apoptosis of PC-3 cells was analyzed by flow cytometry（ FCM）. Western blot was used to detect the expression of Bcl-2,Bax,Cleaved Caspase-3 and PARP. Result Compared with control group,the viability of PC-3 cells decreased after treatment with different concentrations of CPT for 24 h,and the cell morphology were changed in a dose-dependent manner. FCM assay indicated that the early apoptosis ratios were increased after treatment with different concentrations of CPT for 24 h. The cell apoptotic rate increased obviously in CPT-treated groups than in control group. The expression of Bcl-2 was decreased,otherwise,the expression of Bax,Cleaved Caspase-3,and 89-k Da PRPP were increased when compared to those in control groups. Conclusion CPT can induce PC-3 cell apoptosis,which might be related with stimulating the Caspase pathway,further activation of substrate PARP.

关 键 词：CPT 细胞增殖 细胞凋亡 PC-3细胞 

分 类 号：R737.25[医药卫生—肿瘤]