胎儿与成人主要器官miRNAs的表达量分析

发布时间：2018-02-25 19:28

本文关键词： miRNA 基因表达基因调控生长发育　出处：《华中科技大学》2011年硕士论文　论文类型：学位论文

【摘要】：microRNA(miRNA)是生物体内类长约21-25个核苷酸的非蛋白编码的单链小分子RNA。它以碱基互补配对的方式结合到靶mRNA的3非翻译区(3UTR),通过剪切、降解靶mRNA或是抑制靶mRNA的翻译,在基因转录后水平上微调基因的表达。大量的研究证明,miRNA在细胞的分化、增殖与凋亡,生物体的生长发育,肿瘤的形成以及多种疾病的发生等多种过程中起着极其重要的基因表达调控作用。研究表明,miRNA的表达具有组织特异性和表达时序性。我们通过检测和分析我们收集到的例成人与例胎儿各七种组织(肝脏组织、肺组织、大肠组织、脾脏组织、肾脏组织、大脑组织、心脏组织)中,miRNA在二者各相应组织中表达含量的异同,来探究其在生物体生长发育过程中的变化趋势并推测相关miRNA的生物学功能。我们将以上十四个组织分别依次采取:小分子RNA的提取,加多聚A尾,用苯酚-氯仿法抽提多聚A尾的小RNA,逆转录,在特定PCR体系下进行逆转录PCR以及实时荧光定量PCR等实验方法来检测miRNA的表达含量。经过相关的统计学分析,我们发现胎儿样本各组织中总miRNAs的表达量比相对应的成人样本中总miRNAs的表达量高。与此同时,在胎儿及成人相对应的组织中表达量排名前十的miRNA的种类也不尽相同。而胎儿样本跟成人样本在相同组织中同种miRNA的表达量上也有很大的不同。通过对miRNAs靶基因的预测分析,我们发现miRNAs的表达是具有协同性的,很多miRNAs通过协同表达,对某种或几种与细胞增殖,组织分化等生长发育相关或关键的基因进行调控。我们推测在胚胎发育时期,miRNA在生长发育过程中所扮演的角色是至关重要的。相较而言,miRNA在正常成人机体中的所发挥的调控作用则并没有胚胎时期那么的显著。
[Abstract]:MicroRNAs miRNAs are nonprotein-encoded single-stranded small molecules encoding about 21-25 nucleotides in vivo. They bind to the three untranslated regions of the target mRNA in the form of complementary pairs of bases, by cutting, degrading the target mRNA, or inhibiting the translation of the target mRNA. A large number of studies have demonstrated the differentiation, proliferation and apoptosis of miRNA in cells, and the growth and development of organisms. Tumor formation and the occurrence of many diseases play an important role in the regulation of gene expression. Studies have shown that the expression of miRNA is tissue specific and sequential. We detected and analyzed seven tissues (liver, lung, large intestine, spleen, kidney) of adult and fetal tissues we collected. In order to explore the change trend of miRNA in the growth and development of organism and to speculate the biological function of related miRNA, the similarities and differences of the expression of miRNAs in the brain and heart tissues were studied. We took the following 14 tissues in turn: small molecular RNA extraction, addition of poly A tail, phenol chloroform extraction of poly A tail small RNAs, reverse transcriptase, reverse transcriptase (RT), reverse transcriptase (RT), reverse transcriptase (RT), reverse transcriptase (RT). Reverse transcription PCR and real-time fluorescence quantitative PCR were used to detect the expression of miRNA in a specific PCR system. Through the statistical analysis, we found that the expression of total miRNAs in fetal samples was higher than that in adult samples. At the same time, The expression of miRNA in fetal and adult tissues was different from that in adult and fetal samples were different from adult samples in the same tissues. By analyzing the target genes of miRNAs, we find that the expression of miRNAs is synergistic. Many miRNAs cells proliferate with one or more of them by co-expression. We speculate that the role of miRNA in embryonic development is crucial in the process of growth and development. In contrast, the role of miRNA in normal adults. The role of regulation is not as significant as the embryonic stage.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R394

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