运用生物信息学方法筛选肾透明细胞癌中差异表达的关键基因

Screening the differentially expressed key genes in renal clear cell carcinoma by bioinformatics method

  • 摘要:
    目的 基于生物信息学方法分析肌动蛋白结合蛋白(ANLN)及其相关分子与肾透明细胞癌(ccRCC)的关系。
    方法 从基因表达综合(GEO)数据库GSE116251和GSE168845数据集中获得ccRCC组织样本和正常肾组织样本表达谱数据,通过R软件筛选出差异表达微小RNA(DEmiRNAs)和差异表达基因(DEGs), 并进行功能分析和信号通路富集分析。分析DEmiRNAs及其靶基因表达水平与患者总生存期的相关性,并选择P值最小的靶基因及其相互作用的miRNA与肿瘤基因组图谱(TCGA)数据库中ccRCC患者的临床特征进行分析。基于人类蛋白质图谱(HPA)数据库,通过免疫组织化学(IHC)方法验证所选基因在ccRCC组织及正常肾脏组织中的蛋白表达。
    结果 R软件共筛选出3个DEmiRNAs和1 610个DEGs。生存分析数据库分析结果显示, ANLN基因及其相互作用的微小RNA-200c-3p(miR-200c-3p)与ccRCC患者的总生存期相关。富集分析结果显示, DEmiRNAs在细胞环腺苷酸(cAMP)受体介导的信号传导、溶酶体、磷酸二酯水解酶活性和细胞骨架蛋白结合等方面具有独特的功能。DEmiRNAs主要富集于6条信号通路,即破骨细胞分化、金黄色葡萄球菌感染、细胞黏附分子、补体和凝血级联、吞噬体和原发性免疫缺陷。TCGA数据库分析结果显示,不同TNM分期患者的ANLN、miR-200c-3p表达水平比较,差异有统计学意义(P < 0.05)。IHC结果显示,与正常肾脏组织相比, ccRCC组织中的ANLN蛋白表达上调。
    结论 miR-200c-3p及其靶基因ANLN可能参与ccRCC的发展,或可成为ccRCC的潜在生物标志物。

     

    Abstract:
    Objective To analyze the relationship of anilin actin binding protein (ANLN) and its related molecules with clear cell renal cell carcinoma (ccRCC) based on bioinformatics method.
    Methods The expression profile data of ccRCC tissue samples and normal kidney tissue samples were obtained from GSE116251 and GSE168845 datasets of Gene Expression Omnibus (GEO) database. The differentially expressed microRNAs (DEmiRNAs) and differentially expressed genes (DEGs) were screened by R software, and functional analysis and signal pathway enrichment analysis were performed. The correlations of the expression levels of DEmiRNAs and their target genes with overall survival of patients were analyzed, and the target genes with the lowest P-value and their interacting miRNA were selected and analyzed with the clinical characteristics of ccRCC patients from the Tumor Genome Atlas (TCGA) database. Based on the Human Protein Map (HPA) database, the protein expression of the selected genes in ccRCC tissues and normal kidney tissues was verified by immunohistochemistry (IHC).
    Results Three differentially expressed DEmiRNAs and 1 610 differentially expressed DEGs were selected by using R software. Survival analysis database results showed that ANLN gene and its interacting microRNA-200c-3p(miR-200c-3p)were related with the overall survival of patients with ccRCC. Enrichment analysis showed that DEmiRNA had unique functions in cell cyclic adenylate (cAMP) receptor mediated signaling, lysosome, phosphodiester hydrolase activity and cytoskeletal protein binding. Furthermore, DEmiRNAs were mainly enriched in 6 pathways including osteoclast differentiation, Staphylococcus aureus infection, cell adhesion molecules, complement and coagulation cascades, phagosome, and primary immunodeficiency. TCGA database analysis showed that there were statistically significant differences in the expression levels of ANLN and mir-200C-3p in patients with different TNM stages (P < 0.05). IHC outcomes showed that ANLN was up-regulated in ccRCC tissues compared with normal kidney tissue.
    Conclusion MicroRNA-200c-3p and its targets gene ANLN may take part in the development of ccRCC and can be potential biomarkers for patients with ccRCC.

     

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