摘要
目的:筛选抑癌基因DAPK特异的甲基化模式并评价其作为白血病诊断分型标志物的价值。方法:通过亚硫酸氢盐测序法(BSP)分析DAPK基因启动子区Cp G岛在正常人外周血白细胞和HL-60、U937及Jurkat白血病细胞株中甲基化的状态及模式,选择特异的甲基化位点应用甲基化特异性PCR法(MSP)在白血病细胞株和患者外周血标本中验证DAPK基因异常甲基化模式诊断白血病的效能。结果:DAPK基因Cp G岛不同细胞基因组甲基化模式图谱显示,在正常细胞基因组中去甲基化程度高于白血病细胞株,各白血病细胞株甲基化模式存在差异,能找到特异的Cp G甲基化位点并应用MSP检测区分HL-60与其它3种细胞,而临床标本中MSP检测可区分急性非淋巴细胞白血病(ANLL)与其他类型白血病,其诊断的灵敏度、特异度和准确度分别为59.1%、100%和82.7%,没有发现MSP甲基化诊断与临床病理分型之间的关系。结论:DAPK基因启动子区异常甲基化模式作为白血病诊断分型的潜在肿瘤标志物之一,丰富了白血病诊断的方法,为今后寻找各类肿瘤甲基化标志物提供了思路并奠定了研究基础。
Objective: To screen the differential methylation patterns of tumor suppressor gene DAPK and evaluate its value as a biomarker for the diagnosis of leukemia. Methods: The methylation status of DAPK gene promoter' s CpG island w as analyzed in the genomes of normal human w hite blood cells and HL-60,U937 and Jurkat cell lines by bisulfite sequencing PCR( BSP). The effectiveness of differential methylation patterns of DAPK gene for diagnosis of leukemia w as verified in the leukemia cell lines and peripheral blood samples by methylation specific PCR( MSP).Results: The methylation pattern of DAPK gene in different cell genomes displayed that the degree of unmethylation in normal cell genome w as higher than that of leukemia cell lines. The differential CpG sites w ere found and could be used to differentiate HL-60 and the other 3 cell lines by M SP. M eanw hile,the differential methylation patterns in clinical specimens could distinguish acute non-lymphocytic leukemia( ANLL) and other types of leukemia by MSP. The diagnostic sensitivity,specificity and accuracy w ere 59. 1%,100% and 82. 7% respectively. No relationship w as found betw een MSP diagnosis results and clinical pathological typing. Conclusion: The differential methylation patterns of DAPK gene as potential tumor biomarker for diagnosis of leukemia can enrich the means of diagnosis of leukemia,provide idea and basis for finding all kinds of tumor's DNA methylation biomarkers in the future.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2016年第3期687-692,共6页
Journal of Experimental Hematology
