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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 effect of DNMT on the expression of cancer-testis antigen in K562 cells and its mechanism. METHODS: K562 cells were transfected with siRNA targeting different members of the DNMT family, respectively. The levels of DNMT and cancer-testis antigen were detected by RT-PCR. Methylation-specific PCR (MSP) Gene promoter methylation status. Results: After siRNA interference, the expression of DNMT1, DNMT3a and DNMT3b in K562 cells were significantly decreased, and the promoter region of CT10 in cancer-testis was demethylated. However, the non-methylated MAGE-A1 No changes have occurred in the promoter region. The expression of MAGE-A1 and SSX-1 was up-regulated in K562 cells of DNMT group, but not in CT10, PRAME and CT9. However, the expression of NY-ESO-1, HCA587 and HCA661 had no effect. CONCLUSIONS: In K562 cells, interference with DNMT can demethylate the promoter region of some cancer-testis antigen genes, resulting in the re-expression or increased expression of the corresponding cancer-testis antigen molecule.