3种白血病细胞株中P73、DAPK、O^6 MGMT和CDH1基因启动子区CpG岛甲基化模式分析  

Analysis of methylation patterns of CpG islands in the promoter region of P73,DAPK,O^6 MGMT and CDH1 genes from three kinds of leukemia cell lines

作　　者：叶松山[1] 侯俊然[1] 邱耕[2] 毛秉豫[1] 卞华[1] 杨雷[1] 

机构地区：[1]南阳理工学院张仲景国医学院,河南南阳473004 [2]中山大学达安基因股份有限公司

出　　处：《山东医药》2016年第23期23-26,111,共4页Shandong Medical Journal

摘　　要：目的分析3种白血病细胞株中P73、DAPK、O6MGMT和CDH1基因启动子区Cp G岛甲基化模式。方法运用亚硫酸氢盐测序法分析P73、DAPK、O6MGMT和CDH1抑癌基因启动子区Cp G岛在健康人外周血白细胞(正常对照)和U937、HL-60及Jurkat白血病细胞株中甲基化状态,绘制其甲基化图谱,并应用甲基化特异性PCR法在不同细胞中进行验证。结果正常对照、U937、HL-60及Jurkat基因组甲基化率分别为7.26%(238/3 279)、61.33%(1 813/2 956)、89.35%(3 090/3458)、58.10%(1 342/2 310),白血病细胞株与正常对照比较,P均<0.01。P73基因可鉴别正常细胞和肿瘤细胞,DAPK基因可区分开肿瘤细胞HL-60、U937及Jurkat,O6MGMT可进一步把U937与Jurkat区分开。结论 P73、DAPK、O6MGMT和CDH1基因具有特异的甲基化模式,通过3次MSP检测能将正常细胞和白血病细胞株进行鉴别。Objective To analyze the methylation patterns of CpG islands in the promoter region of P73, DAPK, O6MGMT and CDH1 genes from three kinds of leukemia cell lines. Methods The methylation status and maps of P73, DAPK, 06MGMT and CDH1 gene promoters＇ CpG islands were analyzed in the genome of healthy human white blood cells （control group） , U937, HL-60 and Jurkat cell lines by bisulfite sequencing PCR. The differential methylation sites of each gene were verified in the leukemia cell lines by methylation specific PCR. Results The methylation rates of the control group, U937, HL-60 and Jurkat genomes were 7.26% （238/3 279）, 61.33% （1 813/2 956）, 89.35% （3 090/3 458） and 58.10% （1 342/2 310）, respectively. Significant difference was found between the leukemia cell lines and normal control group, all P 〈 0.01 respectively. P73 gene could distinguish between normal and tumor cells, DAPK gene could be used to separate the tumor cells HL-60, U937 and Jurkat; and O6MGMT gene could discriminate U937 and Jurkat further. Conclusion There were specific methylation patterns of P73, DAPK, 06 MGMT and CDH1 genes. The normal cells and leukemia cell lines were identified through MSP detection thrice.

关 键 词：DNA甲基化 亚硫酸氢盐测序 肿瘤抑制基因 肿瘤标志物 白血病 

分 类 号：R733.7[医药卫生—肿瘤]