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目的探讨 Id4基因甲基化作为检测急性白血病(AL)微量残留病变指标的可行性。方法采用甲基化特异性聚合酶链反应(MS-PCR)技术对细胞系中不同比例的白血病细胞以及正常人、初诊和完全缓解期的 AL 患者骨髓进行 Id4基因甲基化状态检测。结果 MS-PCR 方法可在低于1%白血病细胞中检测到Id4基因甲基化。Id4基因在正常骨髓中呈完全性非甲基化状态,初治急性髓系白血病(AML)和急性淋巴细胞白血病(ALL)患者中甲基化比例分别为84%和86%。Id4基因在完全缓解的 ALL 患者中甲基化比例达60.9%,高于完全缓解状态的 AML 患者。Id4基因甲基化检测阳性的14例 ALL 患者中有8例在12个月内复发,而甲基化检测阴性的9例 ALL 患者仅有1例复发。Id4基因呈甲基化状态的8例 AML 患者中有5例在12个月内复发,而 Id4基因呈非甲基化的20例 AML患者中12个月内仅有2例复发。结论 MS-PCR 检测 Id4基因甲基化有可能作为 AL 微量残留病的检测方法。
Objective To investigate the feasibility of methylation of Id4 gene as an indicator of micro-residual disease in acute leukemia (AL). Methods Methylation-specific polymerase chain reaction (MS-PCR) was used to detect the methylation status of Id4 gene in bone marrow of AL patients with different proportions of leukemia cells in normal cell lines, newly diagnosed and fully remissioned patients. Results The MS-PCR method detected Id4 gene methylation in less than 1% of leukemic cells. The Id4 gene was completely unmethylated in normal bone marrow, with methylation rates of 84% and 86% in patients with newly diagnosed acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL), respectively. Id4 gene in ALL patients with complete remission methylation of 60.9%, higher than the complete remission of AML patients. Eight out of 14 ALL patients with a positive methylation of Id4 gene recurred within 12 months, whereas only 9 of 9 ALL patients with methylation negative recurred. Five of the eight AML patients with a methylated Id4 gene recurred within 12 months, while only 20 of 12 patients with Id4 unmethylated had a relapse within 12 months. Conclusion MS-PCR detection of Id4 gene methylation may be used as a detection method of AL micro-residual disease.