HER2高表达胃癌细胞中miR-4728-3p表达及生物信息学分析
Expression and bioinformatics analysis of miR-4728-3p in gastric cancer cells with HER2 overexpression
目的 探讨HER2高表达的胃癌细胞中miR-4728-3p表达状况及其调控靶基因所涉及的信号通路,旨在初步分析miR-4728-3p在胃癌中的作用机制及其与HER2的相关性.方法 首先采用Real-time PCR技术检测胃癌细胞株NCI-N87中miR-4728-3p及HER2 mRNA,然后通过miRanda、DIANA-microT、Targetscan和Targetmine软件预测miR-4728可能的靶基因,并通过DAVID数据库进行靶基因功能富集分析及信号转导通路富集分析.结果 miR-4728-3p及HER2 mRNA在胃癌细胞中均高表达,二者呈正相关(r=0.990,P=0.000).miR-4728-3p预测靶基因共有54个,靶基因功能主要富集于转录活性调节(P=0.014)、DNA结合(P=0.019)及蛋白质磷酸化氨基酸结合(P=0.036)等分子功能,RNA聚合酶Ⅱ启动子的转录调控(P=0.001)、转录调控(P=0.003)及细胞内信号级联(P=0.021)等生物学过程以及轴突部分(P=0.008)等细胞组分上;信号转导通路则主要富集于肿瘤通路(P=0.013)和卵母细胞减数分裂通路(P=0.041).结论 miR-4728-3p在HER2高表达的胃癌细胞中表达上调,二者表达呈正相关;生物信息学分析提示miR-4728-3p通过调控其靶基因参与胃癌的发生发展.
更多Objective To investigate the signaling pathways involved in the target genes regulated by miR-4728-3p and the expression of miR-4728-3p in gastric cancer cells with overexpression of HER2, preliminary analysis of the mechanism of miR-4728-3p in gastric tumorigenesis and its correlation with HER2.Methods Firstly, we detected the level of miR-4728-3p and HER2 mRNA in gastric cancer cell line NCI-N87 by Real-time PCR.Then, we predicted the target genes of miR-4728 using the miRanda, DIANA-microT, Targetscan and Targetmine software, and performed the function enrichment analysis and pathway enrichment analysis of target genes by DAVID database.Results The expression of miR-4728-3p and HER2 mRNA were increased in gastric cancer cells (r=0.990, P=0.000).The fifty four target genes predicted mainly were enriched in molecular functions, as transcription regulator activity (P=0.014), DNA binding (P=0.019), and protein phosphorylated amino acid binding (P=0.036);Also in the biological process , like regulation of transcription from RNA polymerase Ⅱ promoter (P=0.001), regulation of transcription (P=0.003) and intracellular signaling cascade (P=0.021), and in cell components such as axon part(P=0.008).The signaling pathways mainly were enriched in Pathways in cancer (P=0.013) and Oocyte meiosis (P=0.041).Conclusion The expression of miR-4728-3p was up-regulated in gastric cancer cells with overexpression of HER2, and the positive correlation was found between them.Bioinformatics analysis suggested that miR-4728-3p participate in the development of gastric cancer by regulating its target genes.
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