Dubayová Katarína, Kožlejová Zuzana, Vašková Judita, Čakanová Gladys, Kiktavá Mária, Guman Tomáš, Sabo Ján, Karabinos Anton
Klin Onkol. 2018 Spring;31(3):200-206. doi: 10.14735/amko2018200.
The study investigated FLT3 gene mutations in patients from eastern Slovakia using a simple molecular method.
We analyzed 141 patients with primary acute myeloid leukemia (AML) and 8 patients with AML that developed from myelodysplastic syndrome (MDS) who were aged 19-81 years. DNA isolated from peripheral blood and/or bone marrow was analyzed by PCR. FLT3 internal tandem duplication (FLT3-ITD) was detected by amplification of exons 14 and 15. Point mutations in the FLT3 tyrosine kinase domain (FLT3-TKD) were detected by digesting the PCR product of exon 20 with the restriction endonuclease EcoRV. Fragments were separated electrophoretically. PCR products of the positive samples were also analyzed using a microchip device (Bioanalyzer 2100).
LT3-ITD and point mutations in the FLT-TKD were detected in 19 and 8% of patients, resp. Two patients (1%) harbored both types of mutations. Patients with and without FLT3 mutations were called FLT+ and FLT-, resp. Most FLT3+ patients had no chromosomal aberrations (59%) or harbored the t (15; 17) translocation in PML-RARA (15%). The mortality rate was 33% among FLT3+ patients and 10% among FLT3-patients. Among FLT3+ patients, the mortality rates of patients with FLT3-ITD and point mutations of the FLT-TKD were almost the same. A 77-year-old female patient with both FLT3-ITD and a point mutation in the FLT3-TKD was in remission. The eight patients who developed AML from MDS were assessed separately. Of these, three patients were FLT3+; two patients displayed FLT3-ITD, and one patient harbored a point mutation in the FLT3-TKD. No other genetic aberrations were detected. FLT3+ patients lived for longer than FLT3-patients. These analyses of FLT3 gene mutations in patients from eastern Slovakia are consistent with published data from other databases.
The applied PCR method is reliable, relatively fast, and affordable, and can be used for routine monitoring of FLT3 gene mutations. FLT3 mutations can be verified using a microchip as an alternative to capillary electrophoresis. Key words: acute myelogenous leukemia - DNA - PCR - mutation - FLT3-ITD - FLT3-TKD The study was supported by the European Regional Development grant OPVaV-2009/2.2/05- -SORO (ITMS code: 26220220143). The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical paper Submitted: 19. 10. 2017 Accepted: 15. 2. 2018.
本研究采用一种简单的分子方法，对斯洛伐克东部的患者进行FLT3基因突变检测。
我们分析了141例原发性急性髓系白血病（AML）患者以及8例由骨髓增生异常综合征（MDS）发展而来的AML患者，这些患者年龄在19至81岁之间。通过聚合酶链反应（PCR）分析从外周血和/或骨髓中提取的DNA。通过扩增外显子14和15检测FLT3内部串联重复（FLT3-ITD）。通过用限制性内切酶EcoRV消化外显子20的PCR产物，检测FLT3酪氨酸激酶结构域（FLT3-TKD）中的点突变。片段通过电泳分离。阳性样本的PCR产物也使用微芯片设备（Bioanalyzer 2100）进行分析。
分别在19%和8%的患者中检测到FLT3-ITD和FLT-TKD中的点突变。两名患者（1%）同时携带这两种类型的突变。携带和未携带FLT3突变的患者分别称为FLT+和FLT-。大多数FLT3+患者没有染色体畸变（59%）或携带PML-RARA中的t(1；17)易位（15%）。FLT3+患者的死亡率为33%，FLT-患者的死亡率为10%。在FLT3+患者中，FLT3-ITD患者和FLT-TKD点突变患者的死亡率几乎相同。一名77岁的女性患者同时具有FLT3-ITD和FLT3-TKD中的点突变，目前处于缓解期。对8例由MDS发展而来的AML患者进行了单独评估。其中，3例患者为FLT3+；2例患者表现为FLT3-ITD，1例患者在FLT3-TKD中存在点突变。未检测到其他基因畸变。FLT3+患者的存活时间比FLT-患者长。对斯洛伐克东部患者的这些FLT3基因突变分析与其他数据库发表的数据一致。
所应用的PCR方法可靠、相对快速且经济实惠，可用于FLT3基因突变的常规监测。可以使用微芯片验证FLT3突变，作为毛细管电泳的替代方法。关键词：急性髓性白血病 - DNA - PCR - 突变 - FLT3-ITD - FLT3-TKD 本研究得到欧洲区域发展基金OPVaV-2009/2. /05-SORO（ITMS代码：26220220143）的支持。作者声明他们在研究中使用的药物、产品或服务方面没有潜在的利益冲突。编辑委员会声明该手稿符合ICMJE关于生物医学论文的建议 提交日期：2017年10月19日 接受日期：2018年2月15日
