摘要
目的:探讨抑制甲基转移酶(DNMT)对K562细胞中癌-睾丸抗原表达的影响及其机制。方法:分别采用针对DNMT家族不同成员的siRNA转染K562细胞,,采用RT-PCR检测细胞中DNMT及癌-睾丸抗原的水平表达,并采用甲基化特异PCR(MSP)检测部分癌-睾丸抗原基因启动子的甲基化状态。结果:经siRNA干扰后,K562细胞中DNMT1、DNMT3a和DNMT3b的表达量均明显降低,癌-睾丸抗原CT10的启动子区序列发生了去甲基化,但处于非甲基化状态的MAGE-A1启动子区没有发生任何改变。干扰DNMT组的K562细胞,再表达癌-睾丸抗原CT10、PRAME和CT9,而MAGE-A1、SSX-1的表达上调,但是NY-ESO-1、HCA587和HCA661的表达状况均没有任何影响。结论:在K562细胞中,干扰DNMT可使部分癌-睾丸抗原基因的启动子区发生去甲基化,从而导致相应的癌-睾丸抗原分子的再表达或表达增加。
Objective： To investigate the effects and the underlying mechanisms of inhibiting three kinds of DNA methyltransferases （DNMT1, DNMT3a and DNMT3b） on the expression of cancer/testis antigen （CTA） in K562 cell. Methods： Transient transfection of K562 cell with siRNA was targeted against different members of DNMT family. RT-PCR was 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： The expressions of DNMT1, DNMT3a and DNMT3b declined significantly after siRNA interference in K562 cells. Demethylation occurred in the promoter region of CT10, while the promoter region of MAGE-A1 was in a state of unmethylation. CT10, PRAME and CT9, re-expressed in all transfected cells group; The expressions of MAGE-Aland ssx-1 were up-regulated in transfected cells group; There was not expression of NY-ESO-1 HCA587 and HCA661 in all cells. Conclusions： In K562 cells, DNMT can up-regulate the expression of CTA by demethylated the promoter.
出处
《现代生物医学进展》
CAS
2011年第20期3801-3804,3808,共5页
Progress in Modern Biomedicine
基金
国家自然科学基金资助项目(30771995)
