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目的探讨慢性髓系白血病(CML)患者中let-7a-3启动子的甲基化态势及其临床意义。方法建立实时定量甲基化特异性PCR(RQ-MSP)分别检测25例对照者及52例CML患者骨髓单个核细胞中let-7a-3启动子未甲基化水平。结果 52例CML患者未甲基化let-7a-3启动子(59.6%)为阳性,而对照组仅1例(4%)阳性,两组比较差异有统计学意义(P<0.01)。ROC曲线分析表明,let-7a-3启动子未甲基化作为辅助诊断CML有较好的特异性。未甲基化let-7a-3启动子水平与BCR/ABL融合基因转录水平呈显著正相关(r=0.641,P=0.001),但与患者的白细胞、血小板计数及血红蛋白水平无明显相关性(P>0.05)。在慢性期和加速期let-7a-3未甲基化水平显著高于急变期。结论 let-7a-3基因低甲基化水平随疾病进展而降低。
Objective To investigate the methylation status of let-7a-3 promoter in patients with chronic myelogenous leukemia (CML) and its clinical significance. Methods Real-time quantitative methylation-specific PCR (RQ-MSP) was used to detect the unmethylation level of let-7a-3 promoter in bone marrow mononuclear cells of 25 controls and 52 CML patients respectively. Results 52 cases of CML patients were positive for unmethylation let-7a-3 promoter (59.6%), while only 1 case (4%) was positive for control group. There was significant difference between the two groups (P <0.01). ROC curve analysis showed that the unmethylation of let-7a-3 promoter has better specificity for diagnosing CML. There was a significant positive correlation between the level of unmethylated let-7a-3 promoter and the transcriptional level of BCR / ABL fusion gene (r = 0.641, P = 0.001), but no significant correlation with leukocyte, platelet count and hemoglobin P> 0.05). The lethal level of let-7a-3 in chronic phase and accelerated phase was significantly higher than that in blastic phase. Conclusion The low methylation level of let-7a-3 gene decreases with the progress of the disease.