EVI1基因对儿童急性淋巴细胞白血病致病机制的初步研究

发布时间：2018-02-12 10:44

本文关键词： 儿童急性淋巴细胞白血病 EVI1基因致病机制　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:初步探讨EVI基因在儿童急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)中的致病机制。方法:l、收集2015年5月01日至2016年9月30日就诊于青岛大学附属医院血液儿科的30例新发ALL患儿的骨髓,就诊时患儿年龄均小于16岁,同时收集6例正常儿童捐献的骨髓作为正常对照。提取骨髓样本的总RNA,荧光定量qRTPCR检测EVI1 mRNA的相对表达水平,比较ALL患儿与正常对照之间EVI1mRNA的表达差异。2、选取年龄及病情相匹配的患儿2例,进行EVI1基因(+)及EVI1基因(-)ALL患儿骨髓RNA行转录组基因测序,通过部分基因的差异性表达初步探索EVI1基因在儿童ALL中的致病机制。结果:1、对ALL患儿及正常儿童骨髓细胞的EVIl基因表达进行了检测,患儿正常对照EVI1 mRNA表达水平从17.7到20.1,中位表达水平18.9;ALL患儿EVI1mRNA表达水平从0.75到4096,中位表达水平934.65;ALL和正常对照的EVI1mRNA整体相比也有显著差异(p=0.01)。并从中筛选出年龄性别病情相匹配且RNA标本浓度适合转录组基因测序的标本;2、EVI1(+)ALL患儿EVI1基因有一长度为638bp的转录版本,该转录版本在EVI1(-)的患儿中未见表达;3、将两样本差异表达基因数目进行统计,EVI1阳性ALL患儿较EVI1阴性ALL患儿共有1387个基因表达上调,1097个基因表达下调。将上调基因进行分析,发现a.造血干细胞相关且被EVI1基因调控转录的基因包括GATA2、BCL-xL、C/EBPα;b.伴有EVI1阳性的ALL中粒系相关基因表达明显上调,如MPO及CD34、CD13、CD33、CD116、CD114、CD124、CD125、CD126及CD11b;c.EVI1阳性ALL标本中ECM受体表达明显升高。结论:1、ALL患儿中EVI1基因表达量比正常骨髓表达量明显升高。2、EVI1(+)ALL患儿EVI1基因有一长度为638bp的转录版本,该转录版本在EVI1(-)的患儿中未见表达;3、EVI1阳性ALL患儿与EVI1阴性ALL患儿相比共有1387个基因表达上调,1097个基因表达下调。上调基因包括造血干细胞相关且被EVI1基因调控转录的基因(GATA2、BCL-xL、C/EBPα)、粒系相关基因(MPO、CD34、CD13、CD33、CD116、CD114、CD124、CD125、CD126及CD11b)及ECM受体表达相关基因。
[Abstract]:Objective: to explore the pathogenetic mechanism of EVI gene in acute lymphoblastic leukemia (ALL) in children. Methods: bone marrow of 30 newly diagnosed ALL children from May 1st 2015 to September 30th 2016 in blood pediatrics of affiliated hospital of Qingdao University were collected. All the children were younger than 16 years of age at the time of visit. Bone marrow donated from 6 normal children was collected as normal control. The total RNAs of bone marrow samples were extracted and the relative expression of EVI1 mRNA was detected by fluorescence quantitative qRTPCR. The difference of EVI1mRNA expression between children with ALL and normal controls was compared. Two children with matched age and condition were selected to carry out EVI1 gene () and EVI1 gene RNA transcriptional gene sequencing in bone marrow of all children. The pathogenetic mechanism of EVI1 gene in children with ALL was preliminarily explored by differential expression of some genes. Results the expression of EVIl gene in bone marrow cells of children with ALL and normal children was detected at 1: 1. The expression level of EVI1mRNA in normal controls ranged from 17.7 to 20.1, and the median expression level of EVI1mRNA in children with all was from 0.75 to 4096.The median expression level of 934.65 all was also significantly different from that of normal controls. There was a 638bp transcriptional version of the EVI1 EVI1 gene in all children, and the concentration of RNA samples was suitable for the sequencing of transcriptome genes. The number of differentially expressed genes in two samples of EVI1 + ALL patients was statistically analyzed, compared with that of EVI1 negative ALL children, 1 387 genes were up-regulated and 1 097 genes were down-regulated, and the up-regulated genes were analyzed. It was found that the genes related to hematopoietic stem cells and transcribed by EVI1 genes, including GATA2BCL-xLU C / EBP 伪 B. The expression of granulocyte related genes in ALL with EVI1 positive was significantly up-regulated. If the expression of ECM receptor was significantly increased in MPO and CD34, CD13, CD113, CD114, CD124, CD125, CD126 and CD11btc. EVI1 positive ALL specimens, the expression of EVI1 gene was significantly higher in children with 1: 1 all than that in normal bone marrow. [conclusion] there is a 638 BP transcriptional version of EVI1 gene in all children, and the expression of EVI1 gene in all children is significantly higher than that in normal bone marrow. A total of 1 097 genes were down-regulated in children with EVI1 positive ALL and EVI1 negative ALL, including hematopoietic stem cells related to hematopoietic stem cells and transfered by EVI1 gene. These genes were associated with the expression of ECM receptor and ECM receptor genes, such as GATA2, BCL-xLU C / EBP 伪, granulocyte associated gene, MPO4, CD13, CD36, CD116, CD114, CD124, CD125, CD126 and CD11b).
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.71

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