5-杂氮-2′-脱氧胞苷对鼻咽癌细胞株细胞周期及14-3-3σ基因甲基化的影响  

Influence of 5-aza-2′-deoxycytidine on cell cycle of nasopharyngeal carcinoma cell lines and methylation of 14-3-3σ gene

作　　者：谭双香[1] 易红[1] 汤参娥[1] 陈主初[1] 肖志强[1] 

机构地区：[1]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙410008

出　　处：《山西医科大学学报》2008年第6期536-540,563,共6页Journal of Shanxi Medical University

基　　金：教育部跨世纪优秀人才培养计划基金资助项目(教育部科技函[2002]48);芙蓉学者特聘教授科学研究基金资助项目(湘教通[2007]362号);湖南省科技重点科研基金资助项目(06SK2004)

摘　　要：目的探讨5-杂氮-2′-脱氧胞苷(5-aza-2dC)对鼻咽癌细胞株CNE1、CNE2、5-8F、6-10B增殖、凋亡和细胞周期分布的影响,并进一步分析5-aza-2dC处理对14-3-3σ基因甲基化状态的影响。方法用不同浓度5-aza-2dC处理鼻咽癌细胞,MTT实验观察细胞生长曲线,用流式细胞仪检测细胞周期分布和凋亡比例,用甲基化特异性PCR(MSP)分析14-3-3σ基因甲基化状态。结果5-aza-2dC对4株细胞均有生长抑制作用,CNE1和6-10B细胞对药物敏感性高于CNE2和5-8F。5-aza-2dC处理使4株细胞凋亡率出现剂量依赖性的增加,CNE1和6-10B细胞对药物敏感性高于CNE2和5-8F。经5-aza-2dC处理后,CNE1、CNE2、6-10B细胞出现不同程度的G0/G1期阻滞,而5-8F表现为G0/G1期和G2/M期双期阻滞。不经5-aza-2dC处理情况下4株细胞14-3-3σ基因均存在不同程度的甲基化,5-aza-2dC处理能够使14-3-3σ基因去甲基化。结论5-aza-2dC具有抗鼻咽癌作用,抑制作用强弱可能与鼻咽癌分化程度和转移潜能有一定关系,其机制可能与使14-3-3σ等抑瘤基因去甲基化有关。Objective To explore the influence of 5-aza-2′-deoxycytidine（5-aza-2dC）on the proliferation,apoptosis and cell cycle of four nasopharyngeal carcinoma cell lines CNE1,CNE2,5-8F and 6-10B,and to investigate the influence of 5-aza-2dC on methylation of 14-3-3σ gene. Methods Nasopharyngeal carcinoma cell lines were treated with different concentrations of 5-aza-2dC.MTT assay was used to detect the cell growth.Flow cytometry was used to determine cell cycle and apoptosis.Methylation specific PCR was used to evaluate hypermethylation status of 14-3-3σ gene. Results Proliferation of CNE1,CNE2,5-8F,6-10B were all inhibited by 5-aza-2dC,and CNE1 and 6-10B were more sensitive than CNE2 and 5-8F.The apoptosis increased dose-dependently after 5-aza-2dC treatment in all four cell lines,and CNE1 and 6-10B were more sensitive than CNE2 and 5-8F.CNE1,CNE2 and 6-10B were blocked in G0/G1 phase,while 5-8F was blocked in G0/G1 phase and G2/M phase after 5-aza-2dC treatment.Methylation of 14-3-3σ gene was found in different degree in four nasopharyngeal carcinoma cell lines before 5-aza-2dC treatment.After 5-aza-2dC treatment,de-methylation of 14-3-3σ gene increased in a dose-dependent manner. Conclusion 5-aza-2dC has the effect of anti-nasopharyngeal carcinoma,and its effect may be related with tumor differentiation and metastasis potency,which could be related to de-methylation of tumor-suppress gene such as 14-3-3σ.

关 键 词：5-杂氮-2′-脱氧胞苷 鼻咽肿瘤 细胞周期 凋亡 甲基化 

分 类 号：R739.6[医药卫生—肿瘤]