DAPK启动子区异常甲基化模式在白血病诊断分型中的价值分析

Analysis of the value of abnormal methylation pattern in the promoter region of DAPK in the diagnosis of leukemia

目的:探讨抑癌基因DAPK启动子区异常甲基化模式作为潜在肿瘤标志物在白血病诊断分型中的价值。方法:采取亚硫酸氢盐测序法(BSP)对白血病细胞株和正常人外周血白细胞中抑癌基因DAPK启动子的异常甲基化模式进行分析;利用甲基化特异性PCR法(MSP)检验DAPK基因异常甲基化模式对于白血病诊断的效能。结果:Jurkat、U937、HL-60 3种细胞株的甲基化水平和正常细胞株比较差异有统计学意义(χ~2=90.736,P<0.05;χ~2=67.493,P<0.05;χ~2=753.284,P<0.05),提示肿瘤细胞的甲基化水平远高于正常细胞,且HL-60细胞株中的DAPK基因甲基化水平显著高于Jurkat和U937细胞株。ANLL用MSP甲基化检测时特异度、准确度和灵敏度分别为100%(61/61)、82.9%(87/105)和58.7%(27/46),未发现白血病病理分型和MSP诊断之间的关系。结论:抑癌基因DAPK启动子区异常甲基化模式作为一种潜在肿瘤标志物,极大丰富了临床诊断白血病的方法,对于白血病的诊断分型有重要的临床意义。

Objective：To investigate the value of abnormal methylation pattern of DAPK promoter region in the promoter region of tumor suppressor gene as a potential tumor marker in the diagnosis of leukemia.Method：Take the sulfurous acid hydrogen salt sequencing（BSP）on leukemia cell lines and normal human peripheral blood leukocytes in inhibiting cancer gene DAPK promoter abnormal methylation patterns were analyzed;with a methylation specific PCR（MSP）inspection and DAPK gene methylation mode for performance diagnosis of leukemia.Result：Jurkat,U937,HL-60 three cell lines of methylation and significant difference compared with the normal cell lines（χ~2= 90.736,P0.05;χ~2= 67.493,P0.05;χ~2= 753.284,P0.05）,indicating the level of methylation of tumor cells is much higher than normal cells,and HL 60 cell lines of DAPK gene methylation level was significantly higher than that of Jurkat and U937 cell lines.ANLL with MSP methylation detection specific degrees,accuracy and sensitivity was 100%（61/61）,82.9%（87/105）and 58.7%（27/46）,leukemia are not found in the relationship between the pathological classification and MSP diagnosis.Conclusion：As a potential tumor marker,the abnormal methylation pattern of DAPK promoter region of tumor suppressor gene is a potential tumor marker,which greatly enriches the methods of clinical diagnosis of leukemia,and has a very important clinical significance for the diagnosis of leukemia.