CDH13基因在食管癌细胞株EC1和EC109中的甲基化研究

摘要

目的:探讨食管癌细胞中CDH13基因启动子区甲基化状态,以及5-杂氮-2'-脱氧胞苷(5-Aza-CdR)作用对CDH13基因甲基化状态及其表达的影响.方法:5-Aza-CdR处理前后分别采用甲基化特异性PCR(MSP)检测食管癌细胞EC1和EC109中CDH13基因启动子区甲基化状态.westem blot检测CDH13蛋白表达水平的变化.结果:CDH13基因在EC1中呈半甲基化,在EC109中呈完全甲基化.5-Aza-CdR可逆转食管癌细胞中cDH13基因甲基化,恢复其蛋白的表达.结论:cDH13基因异常甲基化可能是其在食管癌中失活的重要方式,应用5-Aza-CdR可逆转甲基化状态.并恢复CDH13表达.%Objective To investigate the methylation status of CDH13 promoter region in human esophageal carcinoma cell lines EC1 and EC109, and the effect of 5-azacytidine-2'-deoxycytidines (5-Aza-CdR) on the methylation status and expression of CDH13. Methods The expression level of CDH13 was measured by Western blot and the methylation status of CDH13 was analyzed by methylation-specific PCR ( MSP) , separately before and after treatment with 5-Aza-CdR. Results The CDH13 gene was hypermethylated in EC109 cells but semi-methylated in EC1 cells. After treatment with 5-Aza-CdR, CDH13 gene methylation was reversed. and the gene expression was strongly upregulated in both cell types. Conclusions The abnormal methylation in promoter region is an important mechanism of CDH13 gene inactivation in human esophageal carcinoma cells. 5-Aza-CdR can effectiveIy reverse such methylation and increase the expression of CDH13.