本文選題：肝細胞癌　切入點：微陣列分析　出處：《重慶醫(yī)科大學》2017年碩士論文

【摘要】：第一部分肝細胞癌中關鍵基因與micro RNA的鑒別和交互作用的生物信息學分析目的:鑒別促進HCC發(fā)生發(fā)展的關鍵基因和miRNA及其潛在的分子機制。方法:從GEO數(shù)據(jù)庫分別下載包含mRNA和miRNA表達譜的微陣列數(shù)據(jù)集GSE22058,GSE25097和GSE57958,并使用GEO2R進行癌及癌旁組織基因差異性分析。使用DAVID數(shù)據(jù)庫對差異基因進行功能和通路富集分析。使用Cytoscape軟件構建PPI網(wǎng)絡。最后,使用miRDB預測差異表達的micro RNA的靶基因。結果:通過分析得到115個DEGs,其中包括52個上調基因和63個下調基因。功能和通路富集分析提示上調基因主要富集于染色體分離和細胞分裂過程,而下調基因主要參與補體激活、蛋白激活級聯(lián)、羧酸代謝過程、含氧酸代謝過程及免疫反應。通過PPI網(wǎng)絡分析篩選18個核心基因,其中ESR1與FOXO1、CXCL12和GNAO1具有密切的相互作用。同時,分析得到64個DEMs,其中hsa-mir-18a和hsa-mir-221等5種miRNA可能對ESR1有靶向調控作用。結論:ESR1及CXCL12等DEGs與has-mir-221等DEMs的相互作用可能在HCC的發(fā)生發(fā)展中起重要作用,這可能為HCC患者的診斷和治療提供新的線索。第二部分EDNRB基因對肝癌細胞株SMMC-7721及Huh7的遷移和侵襲能力的影響目的:探討EDNRB基因對人肝癌細胞遷移及侵襲能力的影響。方法:應用實時熒光定量PCR和Western blot法檢測EDNRB基因mRNA和蛋白在人肝癌細胞株(SMMC-7721和Huh7),人肝細胞株LO2以及HCC組織及其癌旁的相應組織中的表達情況。通過慢病毒轉染構建穩(wěn)定過表達EDNRB基因的細胞株SMMC-7721-E和Huh7-E,應用CCK-8法及Transwell小室法分別檢測EDNRB基因對肝癌細胞增殖、遷移及侵襲能力的影響。結果:EDNRB mRNA在SMMC-7721和Huh7細胞中低于LO2細胞;HCC組織中EDNRB mRNA的表達水平低于相匹配的癌旁組織(p0.01)。SMMC-7721和Huh7細胞中EDNRB蛋白表達低于LO2細胞;HCC組織中EDNRB蛋白的表達低于其相匹配的癌旁組織(p0.01)。EDNRB基因過表達對SMMC-7721和Huh7細胞的增殖無顯著影響(p值均0.05),但能抑制其遷移和侵襲(p值均0.01)。結論:HCC中EDNRB基因的表達顯著下調。EDNRB基因可抑制人肝癌細胞的遷移及侵襲。
[Abstract]:Part I: bioinformatics analysis of the identification and interaction between key genes and micro RNA in hepatocellular carcinoma objective: identification of key genes, miRNA and their potential molecular mechanisms promoting the development of HCC. Methods: from the GEO database. Do not download the microarray datasets GSE22058, GSE25097 and GSE57958, which contain mRNA and miRNA expression profiles, and use GEO2R to analyze the gene diversity of cancer and adjacent tissues. Use the DAVID database to analyze the function and pathway enrichment of differentially expressed genes. Construct the Cytoscape software. Build the PPI network. Finally, MiRDB was used to predict the target genes of differentially expressed micro RNA. Results: 115 DEGs were obtained, including 52 up-regulated genes and 63 down-regulated genes. Functional and pathway enrichment analysis showed that up-regulated genes were mainly enriched in chromosomes. Separation and cell division, The down-regulated genes are mainly involved in complement activation, protein activation cascade, carboxylic acid metabolism, oxygen-containing acid metabolism and immune response. Eighteen core genes were screened by PPI network analysis, in which ESR1 interacted closely with FOXO1CXCL12 and GNAO1. Among 64 demss, five kinds of miRNA, such as hsa-mir-18a and hsa-mir-221, may have targeted regulation on ESR1. Conclusion the interaction of DEGs such as CXCL12 and DEGs with has-mir-221 and DEMs may play an important role in the occurrence and development of HCC. This may provide a new clue for the diagnosis and treatment of HCC patients. Part two: the effect of EDNRB gene on the migration and invasion of hepatoma cell line SMMC-7721 and Huh7 objective: to explore the effect of EDNRB gene on the migration and invasion of human hepatoma cells. Methods: real-time fluorescence quantitative PCR and Western blot were used to detect the expression of EDNRB gene mRNA and protein in human hepatoma cell lines SMMC-7721 and Huh7, LO2, HCC and their adjacent tissues. Cell lines SMMC-7721-E and Huh7-E, which stably expressed EDNRB gene, were constructed by staining. The proliferation of hepatoma cells induced by EDNRB gene was detected by CCK-8 assay and Transwell chamber assay, respectively. Results the expression of EDNRB mRNA in SMMC-7721 and Huh7 cells was lower than that in LO2 cells, and the expression of EDNRB protein in Huh7 cells was lower than that in LO2 cells. The overexpression of white protein was lower than that of matched paracancerous tissues. The overexpression of EDNRB gene had no significant effect on the proliferation of SMMC-7721 and Huh7 cells, but could inhibit its migration and invasion. Conclusion the expression of EDNRB gene can be significantly down-regulated in SMMC-7721 and Huh7 cells. Conclusion\\\; It can inhibit the migration and invasion of human hepatoma cells.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.7

【參考文獻】

相關期刊論文 前2條

1 安湘杰;李艷秋;屈曉鶯;張晶;張凌云;王明;朱里;陳思遠;陳宏翔;涂亞庭;周育文;黃長征;;Silencing Endothelin-3 Expression Attenuates the Malignant Behaviors of Human Melanoma Cells by Regulating SPARC Levels[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2013年04期

2 ;Roles of Chemokine Receptor 4 (CXCR4) and Chemokine Ligand 12 (CXCL12) in Metastasis of Hepatocellular Carcinoma Cells[J];Cellular & Molecular Immunology;2008年05期

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本文編號：1668494