TIMP3、E-Cadherin基因在食管癌细胞系EC1和EC9706中的甲基化及蛋白表达  

Methylation and protein expression of the TIMP-3 and E-cadherin genes in human esophageal carcinoma cell lines EC1 and EC9706

作　　者：赵璇[1] 李沛[1] 马俊芬[1] 赵继敏[1] 杨洪艳[1] 董子明[1] 

机构地区：[1]郑州大学基础医学院病理生理教研室,河南省郑州市450052

出　　处：《世界华人消化杂志》2010年第4期379-383,共5页World Chinese Journal of Digestology

基　　金：河南省高校科技创新人才支持计划基金资助项目;No.2009HAST1T001;郑州大学优秀研究生培育基金资助项目;No.A121~~

摘　　要：目的:观察食管癌细胞系EC1和EC9706中TIMP3、E-Cadherin基因甲基化的水平及蛋白表达,以及去甲基化药物5-Aza-CdR对其的影响.方法:应用甲基化特异性PCR(MSP)和免疫细胞化学的方法,分别检测体外培养的食管癌细胞系EC1和EC9706中TIMP3和E-Cadherin基因甲基化水平及蛋白表达.用5μmol/L5-Aza-CdR处理两种细胞系后,用同样的方法检测其甲基化水平及蛋白表达,观察药物的影响.结果:TIMP3基因在食管癌细胞系EC1和EC9706中均表现为非甲基化,蛋白呈现弱表达;而E-Cadherin基因在EC1中发生甲基化,E C9706中发生半甲基化,蛋白表达均缺失.5-Aza-CdR作用后的两种细胞系中,TIMP3基因仍为非甲基化,而蛋白表达似有所曾强;但E-Cadherin基因甲基化得到了逆转,蛋白表达由原来的不表达,转变为强表达.结论:5-Aza-CdR能够有效逆转E-Cadherin基因甲基化,并使其蛋白恢复表达;TIMP3基因的失活似与甲基化机制无关,5-Aza-CdR对其蛋白表达的恢复作用微弱.AIM： To investigate the methylation and protein expression of the tissue inhibitor of metalloproteinase-3 （TIMP-3） and E-cadherin genes in human esophageal carcinoma cell lines EC1 and EC9706 untreated or treated with the demethylating agent 5-Aza-CdR. METHODS： Methylation-specific PCR （MSP） and imrnunocytochemistry were used to detect the methylation and protein expression of TIMP-3 and E-cadherin genes in EC1 and EC9706 cells untreated or treated with 5 μmol/L of 5-Aza-CdR. RESULTS： The TIMP-3 gene was not methylated, and the TIMP-3 protein was weakly expressed in both EC1 and EC9706 cells. The E-cadherin gene was hypermethylated in EC1 cells but semi-methylated in EC9706 cells. The E-cadherin protein expression was undetectable in both cell types. After treatment with 5-Aza-CdR, the TIMP-3 gene remained non-methylated, and the expression of the TIMP-3 protein was slightly upregulated in the two cell types. In contrast, E-cadherin gene methylation was reversed, and the protein expression was strongly upregulated in both cell types. CONCLUSION： E-cadherin gene methylation occurs in both EC1 and EC9706 cells, and 5-Aza- CdR can effectively reverse such methylation. TIMP-3 gene inactivation seems unrelated to methylation as 5-Aza-CdR can only slightly upregulate the expression of TIMP-3 protein.

关 键 词：食管癌 甲基化 5-AZA-CDR TIMP3 E—Cadherin 

分 类 号：R735.1[医药卫生—肿瘤]