尿毒症患者外周血单个核细胞的超保守区域转录水平研究

发布时间：2018-06-25 10:19

本文选题：T-UCR芯片 + 超保守区域　；参考：《广西师范大学》2014年硕士论文

【摘要】：研究背景：尿毒症(Uremia)并非一种单一疾病,而是由于肾功能衰竭致使代谢产物和其他有毒物质在体内蓄积而起的一种自身中毒综合症,是肾功能衰竭最严重即慢性肾病第V期的表现。尿毒症患者在肾脏替代治疗方面,仍然使用水溶性小分子物质测定作为指标,对其预测性较差。在人、小鼠和大鼠基因中有一段极度的保守长约200-779 bp的序列的非编码基因区域,被称为超保守区域(ultra-conserved regions, UCRs)。由UCR所编码的特殊的长链非编码RNA超保守区域转录子(transcribed ultraconserved regions, T-UCRs),在人类基因组中,它在一些癌症中,如肝癌、结肠癌、神经母细胞瘤等,会改变其表达方式。T-UCRs参与癌症生物学和肿瘤发生,并有可能作为疾病诊断,预后和预测的指标,以及一类新的治疗靶点。目的：文献证实有不少关于尿毒症的并发症以及血液透析后的研究,但很少有研究尿毒症的发病机制,其特异性生物标志物仍然没有找到,尿毒症与T-UCRs表达变化之间可能存在的关联还没有被研究。因此,本研究利用T-UCR芯片技术在基因转录水平上检测分析尿毒症患者的外周血单个核细胞超保守区域转录表达情况,与健康对照组对比,找到差异表达的T-UCRs,并且探讨其与尿毒症的相关性,从而更好的理解尿毒症的发病机制,找到尿毒症的生物标志物。方法：1、利用T-UCR技术来分析15名尿毒症患者和15名健康志愿者的外周血单个核细胞(PBMCs)的超保守区域转录表达水平；2、层次聚类分析尿毒症患者与健康对照组差异表达的T-UCRs的情况；3、GO分析差异表达的T-UCRs的生物学功能情况；4、Pathway分析差异表达的T-UCRs相关的通路情况。结果：经过倍数变化过滤,层次聚类分析,并且和健康对照进行相比,尿毒症患者的潜在T-UCR中有21个T-UCRs表达上调,49个T-UCRs表达下调；对转录范围重叠UCRs的RNA转录子起调控作用的T-UCRs中有311个T-UCRs表达上调,330个T-UCRs表达下调；对UCR-近端基因起调控作用的T-UCRs中有181个T-UCRs表达上调,339个T-UCRs表达下调。GO分析以及pathway分析,挑选出6个表达显著的基因,RP11-369F10.3、HNRPDL、 SNORA74A (p=1.15E-17)、RBFOX2(p=1.865E-22)、SNORD90 (p=0.00211)、FAM24B (p=1.27E-03)作为候选基因,其中RP11-369F1O.3、SNORA74A、SNORD90表达上调,HNRPDL、RBFOX2和FAM24B表达下调。结论：尿毒症患者的超保守区域转录表达水平与健康对照组相比有显著地表达,这些表达或许可以进一步揭示尿毒症的发病机制。这些T-UCRs及候选基因可以作为尿毒症的潜在生物标志物或治疗靶点。
[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.
【学位授予单位】：广西师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R692.5

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