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本实验研究探讨以甲基化特异性聚合酶链反应(MS-PCR)作为白血病微量残留病(MRD)检测方法的可行性。将Id4基因呈完全甲基化的HL-60细胞和Id4基因呈完全非甲基化的Hek937细胞按不同比例混合,实验分为3组:A组为10%HL-60+9%Hek937,B组为1%HL-60+99%Hek937,C组为0.1%HL-60+99.9%Hek937。用MS-PCR方法检测不同白血病细胞比例下Id4基因甲基化情况。结果表明,HL-60细胞仅有155bp的Id4基因甲基化特异性扩增,呈Id4基因完全甲基化;Hek937细胞仅有156bp的Id4基因非甲基化特异性扩增,呈Id4基因完全非甲基化。A、B、C组同时有155bp的Id4基因甲基化和156bp的Id4基因非甲基化特异性扩增,显示Id4基因甲基化表达。结论:MS-PCR方法可从0.1%比例含量的白血病细胞样本中检测到Id4基因甲基化,加之Id4基因甲基化在不同类型白血病中差异不明显,因此用MS-PCR检测Id4基因甲基化状态可以作为一种检测各种类型白血病微量残留病的方法。
This study investigated the feasibility of using methylation-specific polymerase chain reaction (MS-PCR) as a detection method for leukemic minimal residual disease (MRD). HL-60 cells with Id4 gene methylation and Hek937 cells with Id4 gene completely unmethylated were mixed in different proportions. The experiment was divided into three groups: group A was 10% HL-60 + 9% Hek937, B Group 1% HL-60 + 99% Hek937, Group C 0.1% HL-60 + 99.9% Hek937. The methylation status of Id4 gene in different leukemia cells was detected by MS-PCR. The results showed that Id-4 gene of HL-60 cells was only methylated specifically with Id4 gene, and Id4 gene was completely methylated. Hek937 cells only had a non-methylated specific Id4 gene of 156 bp, Non-methylated. The methylation of Id4 gene of 155bp and non-methylation of Id4 gene of 156bp in group A, B and C showed methylation of Id4 gene. Conclusion: Methylation of Id4 gene was detected by MS-PCR in 0.1% leukemia cell samples. The Id4 gene methylation was not significantly different in different types of leukemia. MS detection of Id4 gene methyl The status can be used as a measure of various types of leukemia micro-residual disease method.