本文關(guān)鍵詞： 兒童急性淋巴細(xì)胞白血病 EVI1基因 致病機(jī)制　出處：《青島大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:初步探討EVI基因在兒童急性淋巴細(xì)胞白血病(acute lymphoblastic leukemia,ALL)中的致病機(jī)制。方法:l、收集2015年5月01日至2016年9月30日就診于青島大學(xué)附屬醫(yī)院血液兒科的30例新發(fā)ALL患兒的骨髓,就診時(shí)患兒年齡均小于16歲,同時(shí)收集6例正常兒童捐獻(xiàn)的骨髓作為正常對(duì)照。提取骨髓樣本的總RNA,熒光定量qRTPCR檢測(cè)EVI1 mRNA的相對(duì)表達(dá)水平,比較ALL患兒與正常對(duì)照之間EVI1mRNA的表達(dá)差異。2、選取年齡及病情相匹配的患兒2例,進(jìn)行EVI1基因(+)及EVI1基因(-)ALL患兒骨髓RNA行轉(zhuǎn)錄組基因測(cè)序,通過(guò)部分基因的差異性表達(dá)初步探索EVI1基因在兒童ALL中的致病機(jī)制。結(jié)果:1、對(duì)ALL患兒及正常兒童骨髓細(xì)胞的EVIl基因表達(dá)進(jìn)行了檢測(cè),患兒正常對(duì)照EVI1 mRNA表達(dá)水平從17.7到20.1,中位表達(dá)水平18.9;ALL患兒EVI1mRNA表達(dá)水平從0.75到4096,中位表達(dá)水平934.65;ALL和正常對(duì)照的EVI1mRNA整體相比也有顯著差異(p=0.01)。并從中篩選出年齡性別病情相匹配且RNA標(biāo)本濃度適合轉(zhuǎn)錄組基因測(cè)序的標(biāo)本;2、EVI1(+)ALL患兒EVI1基因有一長(zhǎng)度為638bp的轉(zhuǎn)錄版本,該轉(zhuǎn)錄版本在EVI1(-)的患兒中未見(jiàn)表達(dá);3、將兩樣本差異表達(dá)基因數(shù)目進(jìn)行統(tǒng)計(jì),EVI1陽(yáng)性ALL患兒較EVI1陰性ALL患兒共有1387個(gè)基因表達(dá)上調(diào),1097個(gè)基因表達(dá)下調(diào)。將上調(diào)基因進(jìn)行分析,發(fā)現(xiàn)a.造血干細(xì)胞相關(guān)且被EVI1基因調(diào)控轉(zhuǎn)錄的基因包括GATA2、BCL-xL、C/EBPα;b.伴有EVI1陽(yáng)性的ALL中粒系相關(guān)基因表達(dá)明顯上調(diào),如MPO及CD34、CD13、CD33、CD116、CD114、CD124、CD125、CD126及CD11b;c.EVI1陽(yáng)性ALL標(biāo)本中ECM受體表達(dá)明顯升高。結(jié)論:1、ALL患兒中EVI1基因表達(dá)量比正常骨髓表達(dá)量明顯升高。2、EVI1(+)ALL患兒EVI1基因有一長(zhǎng)度為638bp的轉(zhuǎn)錄版本,該轉(zhuǎn)錄版本在EVI1(-)的患兒中未見(jiàn)表達(dá);3、EVI1陽(yáng)性ALL患兒與EVI1陰性ALL患兒相比共有1387個(gè)基因表達(dá)上調(diào),1097個(gè)基因表達(dá)下調(diào)。上調(diào)基因包括造血干細(xì)胞相關(guān)且被EVI1基因調(diào)控轉(zhuǎn)錄的基因(GATA2、BCL-xL、C/EBPα)、粒系相關(guān)基因(MPO、CD34、CD13、CD33、CD116、CD114、CD124、CD125、CD126及CD11b)及ECM受體表達(dá)相關(guān)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 偽 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 偽, granulocyte associated gene, MPO4, CD13, CD36, CD116, CD114, CD124, CD125, CD126 and CD11b).
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R733.71

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