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氯化镧经Caspase-3途径诱导子宫颈癌细胞凋亡
郭菲1*,赵萍2,缪丽芳3,陆丹3,俞玲芳3,汪泱1*
0
(1. 南昌大学第一附属医院烧伤研究所, 南昌 330006; 2. 江西省科学院人事处,南昌 330029; 3. 南昌大学研究生院医学部,南昌 330006)
摘要:
目的探讨氯化镧体外诱导人子宫颈癌细胞凋亡的作用和分子机制,为探索稀土化合物的药用价值提供理论依据。 方法采用形态学观察、四甲基偶氮唑盐(MTT)比色法测定氯化镧对HeLa细胞的抑制率;流式细胞术分析细胞周期和细胞凋亡率,蛋白质印迹技术分析Caspase-3蛋白表达的改变,并以分光光度法测定其活性。结果形态学分析和MTT法显示氯化镧能够抑制HeLa细胞生长并诱导其凋亡(P<0.05),其作用呈明显的量效关系。细胞周期实验结果说明,氯化镧可令细胞大多阻滞于G1期,并呈一定的浓度依赖性;氯化镧诱导细胞凋亡亦有剂量依赖性,在5 μmol/L剂量下,细胞开始凋亡,随着药物浓度的增加,凋亡率不断增高, 各浓度与对照组相比差异都具有统计学意义(P<0.01)。蛋白质印迹实验显示氯化镧亦可上调活化的Caspase-3蛋白表达水平及活性且呈浓度依赖性。 结论一定浓度的氯化镧可抑制人子宫颈癌HeLa细胞的增殖并通过激活Caspase途径诱导HeLa细胞凋亡。
关键词:    HeLa细胞  细胞凋亡  Caspase-3
DOI:10.3724/SP.J.1008.2011.0541
投稿时间:2011-02-23修订日期:2011-04-02
基金项目:国家自然科学基金 (30960405), 江西省自然科学基金 (2007GZY1132).
Lanthanum chloride induces apoptosis in HeLa cells via Caspase-3 pathway
GUO Fei1*,ZHAO Ping2,MIAO Li-fang3,LU Dan3,YU Ling-fang3,WANG Yang1*
(1. Burns Institute, The First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, China; 2. Department of Human Resources, Jiangxi Academy of Sciences, Nanchang 330006, Jiangxi, China ; 3. Department of Medicine, Postgraduate Department of Nanchang University, Nanchang 330006, Jiangxi, China)
Abstract:
ObjectiveTo explore apoptosis-inducing effect of lanthanum chloride in cervical cancer cells in vitro and the related mechanism, so as to provide a theoretical basis for medical use of rare earth complex. MethodsThe anti-tumor activity of lanthanum chloride against HeLa cells was examined by morphological observation and MTT (3-\[4,5-dimethylthiazol-2-yl\] -2,5-diphenyltetrazolium bromide) assay. Distribution of cell cycle and cell apoptosis were analyzed using flow cytometry (FCM). The expression and activation of Caspase-3 were examined by Western blotting assay and spectrophotometry. ResultsMorphological observation and MTT assay showed that lanthanum chloride significantly induced apoptosis in HeLa cells and inhibited cell growth in a dose-dependent manner(P<0.05). FCM analysis showed that the cell ratio of G0/G1 phase increased and that of S phase decreased significantly with the increase of lanthanum chloride doses. Lanthanum treatment also induced cell apoptosis dose-dependently. The minimum dose of lanthanum chloride to trigger apoptosis was 5 μmol/L, and the apoptosis rate increased in a dose-dependent manner compared with the control group (P<0.01). Western blotting analysis showed that lanthanum chloride up-regulated Caspase-3 expression in a dose-dependent manner. Conclusion Lanthanum chloride can inhibit growth of human cervical cancer HeLa cells and induce cell apoptosis via Caspase pathway.
Key words:  lanthanum  HeLa cells  apoptosis  Caspase-3