肝癌细胞HepG2中p53调控miRNA-3661的生物信息分析与功能验证

发布时间：2018-02-11 20:13

本文关键词： 肝细胞癌 microRNAs 生物信息学阿霉素　出处：《生物技术通报》2017年07期 　论文类型：期刊论文

【摘要】：对已在前期实验中通过Dox诱导肝癌细胞HepG2 DNA损伤发现的受p53调控的hsa-miR-3661进行生物信息学分析,并通过分子生物学实验对其功能进行了验证,为miR-3661在肝肿瘤中的调控机制的研究提供理论基础。获取miR-3661结构与序列信息;预测靶基因,使用DAVID进行miRNA靶基因功能富集分析;分析miR-3661的p53结合位点,通过基因间的相互作用构建调控网络;进行细胞增殖实验验证miR-3661抑制肿瘤功能。结果表明,miR-3661序列保守,启动子区存在p53结合位点,暗示p53与hsa-miR-3661存在直接调控;预测靶基因1 009个,369个显著富集于细胞周期调控、细胞增殖、细胞凋亡等肿瘤相关生物学过程(P0.05),主要参与了癌症信号通路、MAPK信号通路与Erb B信号通路(P0.05);通过268组基因间的相互作用数据构建了p53、hsa-miR-3661和靶基因的调控网络,从系统生物学角度分析了参与多个肿瘤生物进程的关键靶基因;在实验中证实过表达miR-3661可以显著抑制肝癌细胞HepG2的增殖过程(P-value=0.001 46)。miR-3661受p53直接调控,其靶基因显著富集于多种肿瘤相关生物进程与信号通路,过表达miR-3661可显著抑制肝癌细胞增殖。
[Abstract]:The bioinformatics analysis of p53 regulated hsa-miR-3661 in HepG2 DNA damage induced by Dox in previous experiments was carried out, and its function was verified by molecular biological experiments. It provides a theoretical basis for the study of the regulatory mechanism of miR-3661 in liver neoplasms, acquires the structure and sequence information of miR-3661, predicts the target genes, uses DAVID to analyze the function enrichment of miRNA target genes, and analyzes the p53 binding sites of miR-3661. Through the interaction between genes, the regulatory network was constructed, and the inhibition of tumor function by miR-3661 was verified by cell proliferation experiment. The results showed that the sequence of miR-3661 R-3661 was conserved, and the promoter region had p53 binding site, suggesting that p53 and hsa-miR-3661 were directly regulated. It was predicted that 1 009 target genes were significantly enriched in cell cycle regulation and cell proliferation. Apoptosis and other tumor-related biological processes, such as P0.05, are mainly involved in the Erb B signaling pathway and the cancer signaling pathway, and the regulatory network of p53 hsa-miR-3661 and target genes is constructed by the interaction data of 268 groups of genes. The key target genes involved in many tumor biological processes were analyzed from the point of view of systems biology, and it was proved that overexpression of miR-3661 could significantly inhibit the proliferation of HepG2 cells. Its target gene was significantly enriched in many tumor related biological processes and signaling pathways. Overexpression of miR-3661 could significantly inhibit the proliferation of hepatoma cells.
【作者单位】：西南交通大学生命科学与工程学院;
【基金】：中央高校基本科研业务费专项资金(2682016YXZT04) 国家大学生创新性实验计划项目(201610613066)
【分类号】：R735.7

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