他莫昔芬诱导C6胶质瘤细胞凋亡及其作用机制  

Efficiency and Mechanism of Tamoxifen in C6 Glioma Cell Apoptosis

作　　者：田芬 武慧姣[2] 黄锦桃 李朝红[2] 谢富康[2] TIAN Fen;WU Hui-jiao;HUANG Jin-tao;LI Chao-hong;XIE Fu-kang(Reproductive Medicine Center,Xiangya Hospital,Central South University,Changsha 410008,China;Department of Histology and Embryology,Zhongshan School of Medicine,Sun Yat-Sen University,Guangzhou 510080,China)

机构地区：[1]中南大学湘雅医院生殖医学中心,湖南长沙410008 [2]中山大学中山医学院组织学与胚胎学教研室,广东广州510080

出　　处：《中山大学学报（医学版）》2018年第6期811-817,843,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：美国中华医学基金会项目(#98-677);国家自然科学基金青年基金(81701437);广东自然科学基金(2017A030313574)

摘　　要：【目的】探讨他莫昔芬(TAM)对C6胶质瘤细胞凋亡的影响及其可能的作用机制。【方法】(1) TAM经不同浓度、不同时间处理C6胶质瘤细胞后,用台盼蓝染色法检测细胞死亡现象,DAPI组化染色观察凋亡细胞核形态,DNA阶梯法和FACS检测细胞凋亡,Western blot法分析PKCα磷酸化水平。(2)PKCα特异性抑制剂Go6976与TAM联合处理C6胶质瘤细胞后,DNA阶梯法和FACS检测C6胶质瘤细胞凋亡。【结果】(1)TAM能诱导C6胶质瘤细胞死亡,且与TAM浓度、TAM处理时间呈正相关;TAM组可见典型的细胞凋亡的形态学变化(如核固缩)和明显的DNA梯度条带;C6胶质瘤细胞凋亡率和PKCα磷酸化水平具有TAM浓度依赖性和时间依赖性。(2)Go6976与TAM联合处理C6胶质瘤细胞后,可进一步促进TAM诱导的C6胶质瘤细胞凋亡。【结论】TAM能诱导C6胶质瘤细胞凋亡,且PKCα参与介导TAM诱导的C6胶质瘤细胞凋亡。该研究可为临床神经胶质瘤的治疗提供实验依据。【Objective】To investigate the effects of tamoxifen(TAM)on the apoptosis of C6 glioma cells and to explore the possible molecular mechanism.【Methods】C6 glioma cells were treated with different concentrations of TAM at different time points.Cell death was measured by trypan blue staining.Cell apoptosis was evaluated by DAPI staining,DNA ladder and FACS.Phosphorylation of PKCαwas detected by Western blot.After C6 glioma cells were pretreated with or without 10μmol/L Go6976(inhibitor of PKCα)for 1 h followed by different concentrations of TAM(or 0.3%DMSO)exposure for 24 h,we performed agarose gel electrophoresis and FACS to analyze cell apoptosis.【Results】TAM markedly induced cell death of C6 glioma cells,which was positive correlation with the concentrations and treatment time of TAM.The TAM-mediated cell death was mainly apoptosis.Cells treated with TAM displayed typical morphological change of apoptosis such as cell shrinkage,and DNA ladder bands.TAM increased the percentage of apoptotic cells and phosphorylation of PKCαin dose-and time-dependent manners.In addition,Go6976,a specific inhibitor of PKCα,enhanced TAM-induced C6 glioma cells apoptosis.【Conclusion】TAM promoted C6 glioma cell apoptosis.PKCαmediated TAM-induced C6 glioma cell apoptosis.This study would provide useful knowledge for prevention and treatment of human glioma.

关 键 词：C6胶质瘤细胞 细胞凋亡 他莫昔芬 PKCΑ 

分 类 号：R73[医药卫生—肿瘤]