本文選題：T-UCR芯片 + 超保守區(qū)域　； 參考：《廣西師范大學(xué)》2014年碩士論文

【摘要】：研究背景：尿毒癥(Uremia)并非一種單一疾病,而是由于腎功能衰竭致使代謝產(chǎn)物和其他有毒物質(zhì)在體內(nèi)蓄積而起的一種自身中毒綜合癥,是腎功能衰竭最嚴(yán)重即慢性腎病第V期的表現(xiàn)。尿毒癥患者在腎臟替代治療方面,仍然使用水溶性小分子物質(zhì)測(cè)定作為指標(biāo),對(duì)其預(yù)測(cè)性較差。在人、小鼠和大鼠基因中有一段極度的保守長(zhǎng)約200-779 bp的序列的非編碼基因區(qū)域,被稱為超保守區(qū)域(ultra-conserved regions, UCRs)。由UCR所編碼的特殊的長(zhǎng)鏈非編碼RNA超保守區(qū)域轉(zhuǎn)錄子(transcribed ultraconserved regions, T-UCRs),在人類基因組中,它在一些癌癥中,如肝癌、結(jié)腸癌、神經(jīng)母細(xì)胞瘤等,會(huì)改變其表達(dá)方式。T-UCRs參與癌癥生物學(xué)和腫瘤發(fā)生,并有可能作為疾病診斷,預(yù)后和預(yù)測(cè)的指標(biāo),以及一類新的治療靶點(diǎn)。目的：文獻(xiàn)證實(shí)有不少關(guān)于尿毒癥的并發(fā)癥以及血液透析后的研究,但很少有研究尿毒癥的發(fā)病機(jī)制,其特異性生物標(biāo)志物仍然沒(méi)有找到,尿毒癥與T-UCRs表達(dá)變化之間可能存在的關(guān)聯(lián)還沒(méi)有被研究。因此,本研究利用T-UCR芯片技術(shù)在基因轉(zhuǎn)錄水平上檢測(cè)分析尿毒癥患者的外周血單個(gè)核細(xì)胞超保守區(qū)域轉(zhuǎn)錄表達(dá)情況,與健康對(duì)照組對(duì)比,找到差異表達(dá)的T-UCRs,并且探討其與尿毒癥的相關(guān)性,從而更好的理解尿毒癥的發(fā)病機(jī)制,找到尿毒癥的生物標(biāo)志物。方法：1、利用T-UCR技術(shù)來(lái)分析15名尿毒癥患者和15名健康志愿者的外周血單個(gè)核細(xì)胞(PBMCs)的超保守區(qū)域轉(zhuǎn)錄表達(dá)水平；2、層次聚類分析尿毒癥患者與健康對(duì)照組差異表達(dá)的T-UCRs的情況；3、GO分析差異表達(dá)的T-UCRs的生物學(xué)功能情況；4、Pathway分析差異表達(dá)的T-UCRs相關(guān)的通路情況。結(jié)果：經(jīng)過(guò)倍數(shù)變化過(guò)濾,層次聚類分析,并且和健康對(duì)照進(jìn)行相比,尿毒癥患者的潛在T-UCR中有21個(gè)T-UCRs表達(dá)上調(diào),49個(gè)T-UCRs表達(dá)下調(diào)；對(duì)轉(zhuǎn)錄范圍重疊UCRs的RNA轉(zhuǎn)錄子起調(diào)控作用的T-UCRs中有311個(gè)T-UCRs表達(dá)上調(diào),330個(gè)T-UCRs表達(dá)下調(diào)；對(duì)UCR-近端基因起調(diào)控作用的T-UCRs中有181個(gè)T-UCRs表達(dá)上調(diào),339個(gè)T-UCRs表達(dá)下調(diào)。GO分析以及pathway分析,挑選出6個(gè)表達(dá)顯著的基因,RP11-369F10.3、HNRPDL、 SNORA74A (p=1.15E-17)、RBFOX2(p=1.865E-22)、SNORD90 (p=0.00211)、FAM24B (p=1.27E-03)作為候選基因,其中RP11-369F1O.3、SNORA74A、SNORD90表達(dá)上調(diào),HNRPDL、RBFOX2和FAM24B表達(dá)下調(diào)。結(jié)論：尿毒癥患者的超保守區(qū)域轉(zhuǎn)錄表達(dá)水平與健康對(duì)照組相比有顯著地表達(dá),這些表達(dá)或許可以進(jìn)一步揭示尿毒癥的發(fā)病機(jī)制。這些T-UCRs及候選基因可以作為尿毒癥的潛在生物標(biāo)志物或治療靶點(diǎn)。
[Abstract]:Background: Uremia is not a single disease, but an autotoxic syndrome caused by renal failure that accumulates metabolites and other toxic substances in the body. Renal failure is the most serious, chronic nephropathy stage V manifestation. In renal replacement therapy, uremic patients still use water-soluble small molecules as indicators, and their predictability is poor. In human, mouse and rat genes, there is an extremely conserved non-coding region of 200-779 BP, known as ultra-conserved regions (UCRs). (transcribed ultraconserved regions, T-UCRs, a special (transcribed ultraconserved regions, T-UCRs encoded by UCRs, is found in some cancers, such as liver cancer, colon cancer, neuroblastoma, and so on. T-UCRs may be involved in cancer biology and oncogenesis, and may be used as a diagnostic, prognostic and predictor of disease, as well as a new therapeutic target. Objective: there are many studies on the complications of uremia and after hemodialysis, but there are few studies on the pathogenesis of uremia, and its specific biomarkers have not been found. The possible association between uremia and changes in T-UCRs expression has not been studied. Therefore, T-UCR microarray technique was used to detect and analyze the hyperconservative region transcriptional expression of peripheral blood mononuclear cells in uremic patients at the gene transcription level, and compared with the healthy control group. In order to understand the pathogenesis of uremia and find out the biomarker of uremia, we can find the T-UCRs of differential expression, and discuss the correlation between T-UCRs and uremia. Methods T-UCR technique was used to analyze the transcriptional expression of peripheral blood mononuclear cells (PBMCs) in 15 uremic patients and 15 healthy volunteers. 2. Hierarchical cluster analysis of T-UCRs in uremic patients and healthy controls. The biological function of T-UCRs with differentially expressed T-UCRs was analyzed by 3Go. Results: after multiple change filtering, hierarchical cluster analysis, and compared with healthy control, 21 patients with uremia had up-regulated T-UCRs expression and 49 patients had down-regulated T-UCRs expression. There were 311 T-UCRs up-regulated and 330 T-UCRs down-regulated in T-UCRs, 181 T-UCRs were up-regulated, 339 T-UCRs down-regulated by .go and pathway analysis. Six highly expressed genes, RP11-369F10.3HNRPDL, SNORA74A (pN1.15E-17), RBFOX2 (pP1.865E-22), SNORD90 (pP0.00211), FAM24B (pP1.27E-03) were selected as candidate genes, among which the expression of RP11-369F1O.3O. SNORA74AnORD90 up-regulated the expression of HNRPDLpRBX2 and FAM24B down-regulated. Conclusion: the transcriptional expression of hyperconservative regions in uremic patients is significantly higher than that in healthy controls. These expressions may further reveal the pathogenesis of uremia. These T-UCRs and candidate genes can be used as potential biomarkers or therapeutic targets for uremia.
【學(xué)位授予單位】：廣西師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R692.5

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本文編號(hào)：2065581