Laboratory of Molecular Biology, Department of Medical Pathology, Hospital Universitario La Fe, Escuela de enfermería 7ª planta., Avd. Campanar 21, Valencia 46009, Spain.
Ann Hematol. 2012 Jan;91(1):1-7. doi: 10.1007/s00277-011-1234-z. Epub 2011 May 3.
During last years, molecular markers have been increased as prognostic factors routinely screened in acute myeloid leukemia (AML). Recently, an increasing interest has been reported in introducing to clinical practice screening for mutations in the CCAAT/enhancer-binding protein α (CEBPA) gene in AML, as it seems to be a good prognostic factor. However, there is no reliable established method for assessing CEBPA mutations during the diagnostic work-up of AMLs. We describe here a straightforward and reliable fragment analysis method based in PCR capillary electrophoresis (PCR-CE) for screening of CEBPA mutations; moreover, we present the results obtained in 151 intermediate-risk karyotype AML patients (aged 16-80 years). The method gave a specificity of 100% and sensitivity of 93% with a lower detection limit of 1-5% for CEBPA mutations. The series found 19 mutations and four polymorphisms in 12 patients, seven of whom (58%) presented two mutations. The overall frequency of CEBPA mutations in AML was 8% (n = 12). CEBPA mutations showed no coincidence with FLT3-ITD or NPM1 mutations. CEBPA mutation predicted better disease-free survival in the group of patients without FLT3-ITD, NPM, or both genes mutated (HR 3.6, IC 95%; 1.0-13.2, p = 0.05) and better overall survival in patients younger than 65 of this group without molecular markers (HR 4.0, IC 95%; 1.0-17.4, p = 0.05). In conclusion, the fragment analysis method based in PCR-CE is a rapid, specific, and sensitive method for CEBPA mutation screening and our results confirm that CEBPA mutations can identify a subgroup of patients with favorable prognosis in AML with intermediate-risk karyotype.
近年来，分子标志物已被广泛用作急性髓细胞白血病（AML）的常规预后因素。最近，人们越来越关注在 AML 临床实践中引入 CCAAT/增强子结合蛋白α（CEBPA）基因突变筛查，因为它似乎是一个良好的预后因素。然而，在 AML 的诊断工作中，尚未建立可靠的 CEBPA 基因突变评估方法。我们在这里描述了一种基于 PCR 毛细管电泳（PCR-CE）的简单可靠的片段分析方法，用于筛查 CEBPA 突变；此外，我们还介绍了在 151 例中危核型 AML 患者（年龄 16-80 岁）中获得的结果。该方法的特异性为 100%，敏感性为 93%，CEBPA 突变的最低检测限为 1-5%。该系列在 12 名患者中发现了 19 种突变和 4 种多态性，其中 7 名患者（58%）存在两种突变。AML 中 CEBPA 突变的总频率为 8%（n=12）。CEBPA 突变与 FLT3-ITD 或 NPM1 突变无相关性。在没有 FLT3-ITD、NPM 或两者均突变的患者中，CEBPA 突变预测无疾病生存更好（HR 3.6，95%CI 1.0-13.2，p=0.05），在该组年龄小于 65 岁且无分子标志物的患者中，总体生存更好（HR 4.0，95%CI 1.0-17.4，p=0.05）。总之，基于 PCR-CE 的片段分析方法是一种快速、特异、敏感的 CEBPA 突变筛查方法，我们的结果证实，CEBPA 突变可以识别出具有中危核型的 AML 患者中具有良好预后的亚组。
