解放军医学杂志2009，Vol.34Issue(12)：1434-1438,5.
DNA甲基转移酶基因干扰对HepG2细胞癌-睾丸抗原再表达的影响
Effects of interference of DNA methyltransferases in re-expression of cancer/testis antigens in hepatocellular carcinoma cells
摘要
Abstract
Objective To explore the effects and the underlying mechanisms of inhibiting three kinds of DNA methyltransferases (DNMT1, DNMT3a and DNMT3b) on the re-expression of cancer/testis antigen (CTA) in hepatic cells. Methods Transient transfection of HepG2 cells with siRNA was targeted against DNMT1, DNMT3a and DNMT3b (DNMT1+3a+3b), DNMT1 and DNMT3a (DNMT1 +3a), DNMT1 and DNMT3b (DNMTl + 3b), DNMT3a and DNMT3b (DNMT3a+3b), respectively. The other batch of cells was treated with 5-aza-deoxycytidine (5-aza-dC) as positive control. Real-time PCR and Western blotting were used to detect the expressions of DNMTs and CTAsm, and the promoter methylation of partial CTA genes was detected with methylation specific PCR (MSP). Results Expressions of DNMT1, DNMT3a and DNMT3b declined significantly after siRNA interference in HepG2 cells. Two CTAs, CT10 and SSX1, re-expressed in the cells transfected by DNMT1 + 3a and DNMT1 + b. MAGE1 and MAGE3 were equally expressed in all cells despite been transfected or not. Demethylation occurred in the promoter region of CT10, while the promoter region of MAGE1 was in a state of unmethylation. Conclusions In HepG2 cells, CTA promoter can be demethylated via interference of DNMTs, which may lead to the re-expression of CTA that was originally unexpressed due to methylation.关键词
甲基转移酶类/癌-睾丸抗原/癌/肝细胞Key words
methyltransferases/ cancer-testis antigen/ carcinoma/ hepatocellular分类
医药卫生引用本文复制引用
李宏涛,梁后杰,李文梅,肖文华..DNA甲基转移酶基因干扰对HepG2细胞癌-睾丸抗原再表达的影响[J].解放军医学杂志,2009,34(12):1434-1438,5.基金项目
国家自然科学基金资助项目(30572144) （30572144）
