儿童ALL miR-495表达及甲基化水平的研究
发布时间：2018-11-13 06:27
【摘要】：目的:micro RNA-495(mi R-495)在不同肿瘤中可发挥抑癌或致癌的作用,mi R-495的表达水平与启动子区CPG岛甲基化的程度有相关性。儿童ALL中尚未见相关研究。本研究旨在通过检测分析ALL中mi R-495表达水平及甲基化状态的差异,探讨mi R-495在儿童ALL发病中的意义,了解其表达水平及甲基化状态与ALLT/B分型、TEL/AML1等融合基因及诱导缓解疗效的关系。方法:收集80例非超二倍体初发ALL患儿骨髓(bone marrow,BM)和外周血作为实验组,10例免疫性血小板减少性紫癜(immune thrombocytopenia,ITP)儿童骨髓以及31例健康儿童外周血作为对照。提取ALL和ITP及正常人外周血DNA和RNA。对DNA进行重亚硫酸盐处理,对mi RNA进行逆转录处理。采用甲基化敏感性高分辨率溶解曲线分析(methylation sensitivity of high resolution melting curve analysis,MS-HRM)方法对mi R-495基因启动子区CPG岛甲基化水平进行检测;实时荧光定量PCR方法对mi R-495表达水平进行检测。结果:1.mi R-495在ALL组表达水平为1.2800E-4(8.5450E-6,1.5450E-3),显著低于ITP组[9.9500E-2(4.7963E-2,1.4302E-1)]及正常对照组[1.4800E-3(6.1050E-4,4.0850E-3)](P值均≤0.05)。2.ALL中mi R-495高表达组36例,其中B-ALL 35例,T-ALL 1例;低表达组44例,其中B-ALL33例,T-ALL 11例,mi R-495高表达组与低表达组之间B/T构成比差异有统计学意义(P≤0.05)。3.B-ALL中,mi R-495低甲基化组(24例)诱导治疗后其临床危险度分级均未改变,高甲基化组(39例)诱导治疗后其临床危险度分级28例(71.79%)存在危险度调高,mi R-495高甲基化组诱导缓解后其临床危险度上调的比例明显高于低甲基化组(P≤0.05)。4.ALL中mi R-495甲基化水平与表达水平之间无明显相关关系(P0.05)。结论:儿童ALL mi R-495表达水平明显低于ITP及健康儿童对照组;ALL中mi R-495高表达组B-ALL较T-ALL比例显著增加,提示mi R-495在ALL中可能起到肿瘤抑制作用;mi R-495甲基化水平的升高与诱导缓解疗效不佳有关,有望成为对ALL化疗疗效评估的一个生物学指标;儿童ALL中mi R-495甲基化水平与表达水平之间无明显相关关系,mi R-495甲基化可能并非调控其表达水平的主要因素。
[Abstract]:Objective: micro RNA-495 (mi R-495) may play a role in tumor suppressor or carcinogenesis in different tumors. The expression of mi R-495 is correlated with the degree of CPG island methylation in promoter region. No related studies have been found in ALL in children. The purpose of this study was to investigate the significance of mi R-495 in the pathogenesis of ALL in children by detecting and analyzing the difference of mi R-495 expression and methylation status in ALL, and to understand the expression level, methylation status and ALLT/B typing of mi R-495 in children. The relationship between TEL/AML1 fusion gene and induced remission effect. Methods: bone marrow (bone marrow,BM) and peripheral blood of 80 children with non-hyperdiploid ALL were collected as experimental group, 10 children with immunologic thrombocytopenic purpura (immune thrombocytopenia,ITP) and 31 healthy children as control group. Extraction of ALL and ITP and DNA and RNA. from normal Peripheral Blood DNA was treated with heavy sulfite and mi RNA with reverse transcription. The methylation level of CPG island in the promoter region of mi R-495 gene was detected by methylation-sensitive high-resolution dissolving curve analysis (methylation sensitivity of high resolution melting curve analysis,MS-HRM) method. The expression of mi R-495 was detected by real-time fluorescence quantitative PCR. Results: the expression level of 1.mi R-495 in ALL group was 1.2800E-4 (8.5450E-6n 1.5450E-3). It was significantly lower than that in ITP group [9.9500E-2 (4.7963E-2n 1.4302E-1)] and normal control group [1.4800E-3 (6.1050E-4n 4.0850E-3)] (P < 0. 05). There were 36 cases with high mi R-495 expression in 2.ALL. There were 35 cases of B-ALL and 1 case of T-ALL. 44 cases of low expression group, including B-ALL33, 11 T-ALL, mi R-495 high expression group and low expression group, there was significant difference in B / T ratio between the two groups (P 鈮，
本文编号：2328263
[Abstract]:Objective: micro RNA-495 (mi R-495) may play a role in tumor suppressor or carcinogenesis in different tumors. The expression of mi R-495 is correlated with the degree of CPG island methylation in promoter region. No related studies have been found in ALL in children. The purpose of this study was to investigate the significance of mi R-495 in the pathogenesis of ALL in children by detecting and analyzing the difference of mi R-495 expression and methylation status in ALL, and to understand the expression level, methylation status and ALLT/B typing of mi R-495 in children. The relationship between TEL/AML1 fusion gene and induced remission effect. Methods: bone marrow (bone marrow,BM) and peripheral blood of 80 children with non-hyperdiploid ALL were collected as experimental group, 10 children with immunologic thrombocytopenic purpura (immune thrombocytopenia,ITP) and 31 healthy children as control group. Extraction of ALL and ITP and DNA and RNA. from normal Peripheral Blood DNA was treated with heavy sulfite and mi RNA with reverse transcription. The methylation level of CPG island in the promoter region of mi R-495 gene was detected by methylation-sensitive high-resolution dissolving curve analysis (methylation sensitivity of high resolution melting curve analysis,MS-HRM) method. The expression of mi R-495 was detected by real-time fluorescence quantitative PCR. Results: the expression level of 1.mi R-495 in ALL group was 1.2800E-4 (8.5450E-6n 1.5450E-3). It was significantly lower than that in ITP group [9.9500E-2 (4.7963E-2n 1.4302E-1)] and normal control group [1.4800E-3 (6.1050E-4n 4.0850E-3)] (P < 0. 05). There were 36 cases with high mi R-495 expression in 2.ALL. There were 35 cases of B-ALL and 1 case of T-ALL. 44 cases of low expression group, including B-ALL33, 11 T-ALL, mi R-495 high expression group and low expression group, there was significant difference in B / T ratio between the two groups (P 鈮，
本文编号：2328263
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