Department of Biochemistry and Microbiology, Faculty of Pharmacy, Tishreen University, Ministry of High Education, Lattakia, Syria.
Department of Molecular Biology and Biotechnology, Human Genetics Division, Atomic Energy Commission, Damascus, Syria.
Asian Pac J Cancer Prev. 2022 Apr 1;23(4):1387-1395. doi: 10.31557/APJCP.2022.23.4.1387.
Among all types of hematological neoplasms, acute myeloid leukemia (AML) has the highest death rate. Recently, cytogenetic and molecular genetics are crucial in the management, as a consequence of their effect on AML pathogenesis, classification, risk-stratification, prognosis and treatment.
100 Syrian adults with Normal Karyotype (NK) newly diagnosed AML patients were included in this study, all cases confirmed histologically and immunohistochemically. Patients were divided into six subgroups using flow cytometry and cytological results. Polymerase chain reaction (PCR) was performed on exon 11-12 for FMS-like tyrosine kinase-3 internal tandem duplication (FLT3-ITD), exon 12 for Nucleophosmin1 (NPM1), and exon 23 for DNA methyltransferase 3A (DNMT3A) using target primers, the electropherograms were analyzed for gene mutations by comparing with the reference DNA sequence. Data were compared and aligned with different sequences using the NCBI BLAST Assembled Genomes tool.
FLT3-ITD, NPM1 and DNMT3A were detected in 24%, 22 % and 4% patients respectively. M2 subtype had the most frequent incidence of diagnosis in AML. FLT3-ITD mutation patients had the highest mean of death cases, while the DNMT3A mutation patients had the lowest. On the other hand, the highest mean of remission was in patients with NPM1 mutation and the lowest in the carriers of the FLT3-ITD mutation. It was observed that the mean relapsed patients with FLT3-ITD and DNMT3A mutation was 3.4 and 2 months respectively, with no significant differences between (FLT3-ITD and DNMT3A) carriers and non-carriers relapsed. On the contrary, the mean relapsed for NPM1 mutation carriers was 2.4 months with significant statistical differences. The mean survival time for patients with FLT3-ITD and NPM1 mutation was 5.9 months and 5.85 months respectively, with significant correlation. Between it was 5.88 months in DNMT3A patients with no significant differences. Finally, It was noted that the mean event free survival (EFS) of FLT3-ITD mutation patients was 4.818 months and the mean EFS of NPM1 mutation patients was 4.805 months, with significant statistical differences (p<0.05) between the mutation patients and non-mutated patients regarding to EFS, While this mean was not statistically significant in patients carrying DNMT3A mutation.
Patients with FLT3-ITD and NPM1 mutations have the worst prognosis, where the presence of those mutations was significantly related to overall survival (OS) and EFS. Our study reflects that DNMT3A was not an extremely bad prognostic effect as an independent factor. We can declare according to this study that genetic mutation and variants detection could easily be incorporated into the regimen evaluation of AML patients.
在所有类型的血液系统恶性肿瘤中，急性髓系白血病（AML）的死亡率最高。最近，细胞遗传学和分子遗传学在 AML 的发病机制、分类、风险分层、预后和治疗方面具有重要意义。
本研究纳入了 100 例新诊断的具有正常核型（NK）的叙利亚成人 AML 患者，所有病例均经组织学和免疫组织化学证实。根据流式细胞术和细胞学结果，将患者分为六组。使用目标引物通过聚合酶链反应（PCR）检测 FMS 样酪氨酸激酶-3 内部串联重复（FLT3-ITD）、核磷蛋白 1（NPM1）的外显子 12 和 DNA 甲基转移酶 3A（DNMT3A）的外显子 23，通过与参考 DNA 序列比较分析基因突变。使用 NCBI BLAST 组装基因组工具比较和对齐数据，并与不同序列进行比较。
FLT3-ITD、NPM1 和 DNMT3A 在 24%、22%和 4%的患者中分别被检测到。在 AML 中，M2 亚型的诊断发生率最高。FLT3-ITD 突变患者的死亡例数最高，而 DNMT3A 突变患者的死亡例数最低。另一方面，NPM1 突变患者的缓解率最高，FLT3-ITD 突变患者的缓解率最低。观察到 FLT3-ITD 和 DNMT3A 突变患者的平均复发时间分别为 3.4 个月和 2 个月，突变携带者和非携带者之间无显著差异。相反，NPM1 突变携带者的平均复发时间为 2.4 个月，具有显著的统计学差异。FLT3-ITD 和 NPM1 突变患者的平均总生存时间分别为 5.9 个月和 5.85 个月，具有显著相关性。DNMT3A 患者的平均总生存时间为 5.88 个月，无显著差异。最后，我们注意到 FLT3-ITD 突变患者的无事件生存（EFS）平均为 4.818 个月，NPM1 突变患者的 EFS 平均为 4.805 个月，突变患者与非突变患者在 EFS 方面有显著的统计学差异（p<0.05），而在携带 DNMT3A 突变的患者中，这一平均值没有统计学意义。
FLT3-ITD 和 NPM1 突变患者的预后最差，这些突变的存在与总生存（OS）和 EFS 显著相关。我们的研究表明，DNMT3A 作为一个独立的因素并不是一个极其恶劣的预后效应。根据这项研究，我们可以宣称，遗传突变和变异检测可以很容易地纳入 AML 患者的治疗评估。
