MicroRNA-192和-215通过靶基因BIVM参与人胃细胞生物学功能研究

发布时间：2018-02-01 03:38

本文关键词： microRNA-192和-215 胃癌 BIVM　出处：《深圳大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究背景与目的微小RNA(microRNA,miRNA)是一类19-22个核苷酸的内源性非编码小RNA,miRNA的5’端的2-7个核苷酸的“种子序列”可以与靶mRNA的3’端非翻译区互补结合,在转录后水平调控靶基因的表达。1993年,miRNA-lin-4被发现,此后又陆续从线虫、果蝇、家鼠、拟南芥及人类等多种真核生物中发现了这类小分子RNA,据推测在人类染色体上就有1000余种miRNA,目前已有超过700种得到确认。miRNA的编码基因是目前最大的一类调节基因。它在细胞的生长、增殖、发育和凋亡等诸多重要过程中均发挥了关键作用,并与多种肿瘤的发生发展密切相关。大量研究表明,miRNAs在肿瘤中扮演着相当于癌基因或抑癌基因的角色,调节肿瘤细胞的多种重要的生物学行为。胃癌(gastric cancer)是世界上最常见的恶性肿瘤之一;在我国其发病率居各类肿瘤的首位,每年有超过17万人死于胃癌。目前,住院病例中90%为进展期胃癌,尽管针对胃癌的诊断和治疗已经有了长足的进步,但进展期胃癌的预后依然很差,五年生存率长期徘徊在30%左右。近年来已发现胃癌的发生是多基因改变的结果,且已有证据表明miR-21,miR-191,miR-223,let-7a microRNA,miR-106b-25亚群等与胃癌的发生密切相关:let-7 microRNA,miR-155等与胃癌的侵袭与转移密切相关。随着越来越多的miRNA在胃癌中被发现,miRNA在胃癌的基因诊断和治疗中将发挥更为重要的作用。目前,还没有关于BIVM(for basic,immunoglobulin-like variable motif-containing)与胃癌关系的研究。基于本课题组有关胃癌miRNA表达谱的研究,我们初步确定BIVM为研究重点。以BIVM基因为检索词,利用四个常用数据库microRNA.org、Target Scan、Pictar和miRanda进行相互作用miRNAs的预测,挑选出预测值较高(至少三个软件均预测到)的MiR-192和miR-215作为候选miRNAs;基因芯片、Real-time RT-PCR和Western blot结果显示MiR-192和miR-215可能是BIVM相互作用的miRs:荧光素酶报告系统结果显示miR-192和miR-215 inhibitors上调BIVM的后转录水平;然而MiR-192和miR-215 mimics起相反作用。向胃细胞系HFE145、SGC7901、BGC823中转染BIVM-siRNA以降低BIVM的蛋白表达水平来研究BIVM对胃细胞生物学功能的影响。
[Abstract]:Background and objective MicroRNAs miRNAs are a class of 19-22 nucleotides endogenous non-coding small RNA. The "seed sequence" of 2-7 nucleotides at the 5'end of miRNA can be complementary to the 3'-terminal untranslated region of the target mRNA, regulating the expression of the target gene at the post-transcriptional level. 1993. MiRNA-lin-4 was discovered, and RNA was found in a variety of eukaryotes such as nematodes, Drosophila, mice, Arabidopsis thaliana and humans. It is speculated that there are more than 1000 kinds of miRNAs on human chromosomes. At present, more than 700 kinds of miRNAs have been identified as the largest class of regulatory genes. Proliferation, development and apoptosis play a key role in many important processes, and are closely related to the occurrence and development of many kinds of tumors. MiRNAs plays the role of oncogene or tumor suppressor gene in tumors. Gastric cancer is one of the most common malignant tumors in the world. At present, 90% of the hospitalized cases are advanced gastric cancer, although the diagnosis and treatment of gastric cancer has made great progress. However, the prognosis of advanced gastric cancer is still very poor, the long-term survival rate of five years hovering around 30%. In recent years, it has been found that the occurrence of gastric cancer is the result of multiple gene changes, and there is evidence that miR-21. MiR-191 and miR-2223 let-7a microRNA. MiR-106b-25 subsets were closely related to the occurrence of gastric cancer: let-7 microRNA. MiR-155 et al are closely related to the invasion and metastasis of gastric cancer. With more and more miRNA found in gastric cancer. MiRNA will play a more important role in the gene diagnosis and treatment of gastric cancer. At present, there is no BIVM(for basic. Immunoglobulin-like variable motif-containing). Study on the relationship between gastric cancer and gastric cancer. Based on the study of miRNA expression profile of gastric cancer. We preliminarily identified BIVM as the focus of our research. With the BIVM base as the key word, we used four commonly used databases, microRNA.org.Target Scan. Pictar and miRanda were used to predict the interaction of miRNAs. MiR-192 and miR-215 with high predictive value (at least three softwares predicted) were selected as candidates; Gene chip. The results of Real-time RT-PCR and Western blot suggest that MiR-192 and miR-215 may be MIRs that interact with BIVM:. The results of luciferase reporting system showed that miR-192 and miR-215 inhibitors up-regulated the post-transcription level of BIVM. However, MiR-192 and miR-215 mimics had the opposite effect on the gastric cancer cell line HFE145GC7901. The effect of BIVM on the biological function of gastric cells was studied by transfection of BIVM-siRNA in BGC823 to reduce the expression of BIVM protein.
【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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