Department of Hematology, Faculty of Paramedicine, AJA University of Medical Sciences, Tehran, Iran.
Department of Medical Laboratory, Khomein University of Medical Sciences, Khomein, Iran.
Asian Pac J Cancer Prev. 2022 Jul 1;23(7):2185-2190. doi: 10.31557/APJCP.2022.23.7.2185.
Acute myeloid leukemia (AML) is caused by abnormal gene expression following mutations. Many of the mutations in AML lead to gene instability and poor response to treatment. Among these mutations, DNMT3A mutation is exceedingly important due to its major role in methylation and its effect on the expression of other genes. Aberrant methylation due to DNMT3A mutations that mostly occur in exon 23, affects the overall survival (OS) of patients with AML and myelodysplastic syndromes (MDS) showing the importance of identification of these mutations. According to the association of these mutations with short overall survival and disease progression in AML patients, we aimed to investigate DNMT3A gene exon 23 mutations using HRM.
Fifty peripheral blood samples were taken from patients with AML. Mononuclear cells were isolated by ficoll method, and DNA was extracted. Then, mutation detection was detected using the HRM method. Efficacy of the HRM method in mutation detection was compared with direct sequencing method as gold standard.
Mutations in codon 23 of the DNMT3A gene were detected in 5 patients (10%). All of the detected mutations were missense type. A comparison between direct sequencing and HRM analysis demonstrated full concordance of mutation detection.
According to the full consistency between the HRM and direct sequencing methods, HRM is suggested to be adopted as an alternative for the common time-consuming methods in detecting the gene mutations.
急性髓系白血病（AML）是由基因突变导致的异常基因表达引起的。AML 中的许多突变导致基因不稳定和对治疗的反应不良。在这些突变中，DNMT3A 突变由于其在甲基化中的主要作用及其对其他基因表达的影响而极为重要。由于 DNMT3A 突变主要发生在外显子 23 中而导致的异常甲基化，影响 AML 和骨髓增生异常综合征（MDS）患者的总生存期（OS），表明鉴定这些突变的重要性。根据这些突变与 AML 患者总体生存时间短和疾病进展的相关性，我们旨在使用 HRM 研究 DNMT3A 基因外显子 23 突变。
从 AML 患者中采集了 50 份外周血样本。通过 ficoll 法分离单核细胞，并提取 DNA。然后，使用 HRM 方法检测突变。将 HRM 方法在突变检测中的功效与直接测序方法（金标准）进行比较。
在 5 名患者（10%）中检测到 DNMT3A 基因 23 号密码子的突变。所有检测到的突变均为错义型。直接测序和 HRM 分析之间的比较表明突变检测完全一致。
根据 HRM 和直接测序方法之间的完全一致性，建议采用 HRM 作为检测基因突变的常用耗时方法的替代方法。
