文章摘要
梁伟徐兰杨荣秉初明连觅朱蕴兰余培峰徐碧荷王月丹.DNA 甲基转移酶基因干扰对K562 细胞癌- 睾丸抗原表达的影响[J].,2011,11(20):3801-3804
DNA 甲基转移酶基因干扰对K562 细胞癌- 睾丸抗原表达的影响
Effects of Interference of DNA Methyltransferases in Expression ofCancer/Testis Antigens in K562 Cell
  
DOI:
中文关键词: 甲基转移酶  癌- 睾丸抗原  肿瘤  K562 细胞
英文关键词: Methyltransferases  Cancer-testis antigen  Tumor  K562 cell line
基金项目:国家自然科学基金资助项目(30771995)
作者单位
梁伟徐兰杨荣秉初明连觅朱蕴兰余培峰徐碧荷王月丹 北京大学基础医学院免疫学系 
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中文摘要:
      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。
英文摘要:
      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-A1and 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.
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