摘要
目的:观察去甲基化药物5-杂氮-2’-脱氧胞苷(5-Aza-CdR)和5-氟尿嘧啶(5-FU)对食管癌细胞系EC1和EC9706(均存在E-Cadherin甲基化改变和TIMP3基因未发生甲基化改变)中2种基因甲基化状态及蛋白表达的影响。方法:用5-Aza-CdR和5-FU分别作用于EC1和EC9706细胞,应用甲基化特异性PCR检测2种药物处理前后细胞中E-Cadherin基因和TIMP3基因的甲基化状态,应用免疫细胞化学方法检测TIMP3和E-Cadherin蛋白的表达。结果:5-Aza-CdR或5-FU处理后两种食管癌细胞系中TIMP3基因仍为非甲基化;5-Aza-CdR作用后,两种食管癌细胞系中E-Cadherin基因的甲基化状态发生了逆转,而5-FU作用后,E-Cadherin基因的甲基化状态无改变。EC1和EC9706经5-Aza-CdR处理后TIMP3蛋白表达无改变,而经5-FU处理后表达增强(P<0.05),EC1和EC9706经两种药物处理后E-Cadherin蛋白表达均增强(F=116.835、119.628、191.700和210.532,P<0.001)。结论:5-Aza-CdR能够有效逆转EC1和EC9706细胞中E-Cadherin基因甲基化状态,并使其蛋白恢复表达;对TIMP3基因的甲基化状态和蛋白表达影响不大。
Aim：To observe the effects of demethylating drug 5-Aza-2’-deoxycytidine （5-Aza-CdR） and 5-FU on E-Cadherin（with methylation） and TIMP3 （without methylation） gene methylation status and their protein expressions in EC1 and EC9706 cells.Methods：5-Aza-CdR and 5-FU were used respectively to treat EC1 and EC9706 cells.The methylation-specific PCR and immunocytochemical method were used to detect TIMP3 and E-Cadherin gene methylation and protein expression levels,respectively.Results：In EC1 and EC9706 cell lines,after being treated with 5-Aza-CdR or 5-FU,TIMP3 genes remained non-methylation,while the E-Cadherin genes in EC1 with hypermethylation,in EC9706 with semi-methylation had been reversed by 5-Aza-CdR,but there were no changes in methylation status of 2 genes after being treated with 5-FU.However,with 2 drugs treated respectively,E-Cadherin protein expression changed from the weak expression or no expression into strong expression,and TIMP3 had no change with 5-Aza-CdR treated and had strong expression with 5-FU treated（F=191.700,210.532,116.835 and 119.68,P0.001）.Conclusion：The methylation of E-Cadherin in EC1 and EC9706 cells may be the reason causes the inactivation of E-Cadherin gene,while 5-Aza-CdR can reverse the methylation and strengthen the protein expression of E-Cadherin.The inactivation of TIMP3 gene is not associated with the mythylation,and 5-FU can strengthen its protein expression.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2010年第5期719-723,共5页
Journal of Zhengzhou University(Medical Sciences)
基金
河南省高校科技创新人才支持计划基金资助项目2009HAST1T001
郑州大学优秀研究生培育基金资助项目A121
