急性髓系白血病RASSF1A基因启动子区异常甲基化临床意义研究

发布时间：2018-05-21 09:26

本文选题：RASSFA基因 + 甲基化　；参考：《中华肿瘤防治杂志》2017年15期

【摘要】：目的 RASSF1A基因异常甲基化可能参与血液肿瘤的发生,并为微小残留疾病(minimal residual de-sease,MRD)监测、分层、预后评估及靶向治疗提供依据。本研究旨在分析RASSF1A基因启动子区异常甲基化在急性髓系白血病(acute myeloid leukemia,AML)中的临床意义。方法选取2005-01-01-2013-03-01解放军总医院(113例)以及第一附属医院(39例)住院患者和门诊体检患者,共152例AML患者骨髓标本以及15例健康供者骨髓标本纳入本项研究。提取基因组DNA,并进行DNA硫化修饰;设计重亚硫酸盐测序PCR(bisulfite sequencing PCR,BS-PCR)引物以及甲基化特异性PCR(methylation specific PCR,MS-PCR)引物,进行PCR扩增,进而电泳分析以及DNA序列分析。同时对RASSF1A高甲基化组以及低甲基化组的血液学特点、骨髓原始细胞比例、细胞遗传学异常、基因异常、完全缓解率和总生存期进行统计学分析。结果 MS-PCR分析结果显示,RASSF1A基因在15例健康人中呈完全非甲基化状态,在152例AML患者中有38例出现启动子区高甲基化状态,其甲基化阳性率为25%。4例MS-PCR阳性AML患者经BS-PCR测序分析后,显示RASSF1A甲基化率分别为88.2%、85.5%、78.6%和92.7%,而在4例MS-PCR阴性患者RASSF1A基因启动子区甲基化率分别为10%、11.8%、12.7%和6.8%,4例健康供者RASSF1A基因启动子区甲基化率分别为5.0%、9.1%、8.2%和7.3%。进而通过统计学分析发现携带RASSF1A基因高甲基化的AML患者易合并存在ASXL1基因突变或DNMT3A基因突变。携带RASSF1A基因高甲基化的AML患者,其无进展生存期以及总生存期较短。结论 RASSF1A基因启动子区异常甲基化可能参与AML的发生,同时可能为AML分层诊治以及预后评估提供分子理论依据。
[Abstract]:Objective abnormal methylation of RASSF1A gene may be involved in the development of hematologic tumors, and provide evidence for minimal residual de-season MRDs monitoring, stratification, prognostic evaluation and targeted therapy. The purpose of this study was to analyze the clinical significance of abnormal methylation of RASSF1A gene promoter in acute myeloid leukemia. Methods the bone marrow specimens of 152 AML patients and 15 healthy donors were included in this study, including 113 cases of PLA General Hospital and 39 cases of the first affiliated Hospital of PLA General Hospital from January to January 2005. The genomic DNA was extracted and modified with DNA, and the primers for PCR(bisulfite sequencing PCR- BS-PCR and methylation-specific PCR(methylation specific PCR- MS-PCR were designed for PCR amplification, electrophoresis analysis and DNA sequence analysis. At the same time, the hematological characteristics, bone marrow primordial cell ratio, cytogenetic abnormality, gene abnormality, complete remission rate and total survival of RASSF1A hypermethylation group and hypomethylated group were analyzed statistically. Results the results of MS-PCR analysis showed that the RASSF1A gene was completely nonmethylated in 15 healthy persons, 38 of the 152 AML patients had hypermethylation status of promoter region, and the positive rate of methylation was 25.4 MS-PCR positive AML patients. After BS-PCR sequencing analysis, the positive rate of RASSF1A gene was analyzed by BS-PCR sequencing. The results showed that the methylation rate of RASSF1A was 88.2% and 92.7%, respectively, while in 4 MS-PCR negative patients, the methylation rate of the promoter region of RASSF1A gene was 10.11.8% and 12.7%, respectively, and the rate of methylation of RASSF1A promoter was 5.09.1.2% and 7.33% in 4 healthy donors, respectively. Furthermore, it was found that AML patients with hypermethylation of RASSF1A gene were likely to have ASXL1 gene mutation or DNMT3A gene mutation by statistical analysis. In AML patients with hypermethylation of RASSF1A gene, the progression-free survival and total survival were shorter. Conclusion abnormal methylation of RASSF1A promoter region may be involved in the pathogenesis of AML and may provide molecular theoretical basis for AML stratification diagnosis and prognosis evaluation.
【作者单位】：解放军总医院肿瘤中心;解放军总医院第一附属医院放疗科;中日友好医院血液科;
【分类号】：R733.71

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