摘要
目的：探讨肿瘤细胞中JWA基因启动子甲基化的状态及其对基因转录活性的影响。方法用报告基因法检测JWA基因启动子甲基化对基因转录活性的影响，用亚硫酸氢盐测序法检测JWA基因启动子CpG岛内的甲基化位点。用甲基化特异性PCR法（methylation specific PCR，MSP）法检测不同肿瘤细胞株（U937、HL60、K562、NB4）以及在白血病患者外周血单个核细胞中JWA基因启动子甲基化的分布。亚硫氢酸盐测序法检测CpG岛内发生甲基化位点。结果用甲基化酶处理报告基因后JWA基因启动子的转录活性明显下降。用亚硫氢酸盐测序法检测肿瘤细胞株中JWA基因启动子CpG岛未发现甲基化的CpG位点。用MSP法检测U937、HL60、K562、NB4肿瘤细胞和白血病患者外周血单个核细胞的DNA样本均未发现肿瘤细胞启动子甲基化。结论JWA基因启动子甲基化可导致肿瘤细胞JWA基因的转录活性下降，但在所检测的肿瘤细胞及白血病细胞中可能无甲基化现象。
Objective To investigate the effect of methylation status of the 5 [扌入] pG islands in JWA gene and its effect on the activity of transcription. Methods The effect of methylation of promoter on activity of transcription were analyzed by reporter vector, and the frequency of DNA methylation of 4 tumor cell lines and mononuclear cells from leukemia patients were detected by methylation specific PCR （MSP）. The PCR products were cloned and sequenced directly to analyzed methylation sites in CpG island of JWA. Results Methylation of promoter of can decrease The transcription activity of JWA gene markedly decreased in vitro following the treatment of reporter genes by. Sss I. No methylation sites of CpG were found in U937, HL60, K562 and NB4 cell lines and in the mononuclear cells of leukemia patients. Conclusion Methylation of JWA gene promoter may lead to decrease the activity of transcription, but it seems to be likely that the methylation in the DNA of the examined tumor cells as well as in the leukemia cells is absent.
出处
《医药世界》
2006年第11期59-62,共4页
Medicine World
