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肝泡型包虫病侵及胆道患者胆汁外泌体microRNA的检测及生物信息学分析
Measurement and bioinformatics analysis of exosomes microRNAs in bile of hepatic alveolar echinococcosis patients with biliary tract invasion
文章发布日期:2020年08月22日  来源:  作者:王志鑫,苟平,于文昊,等  点击次数:584次  下载次数:36次

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【摘要】:目的 检测肝泡型包虫病侵及胆道患者胆汁中外泌体microRNA(miRNA)的表达情况,探索差异miRNA对靶基因的调控机制。方法 收集自2017年8月-2018年10月就诊于青海大学附属医院的12例肝泡型包虫病患者胆汁样本,观察组(有胆道受侵表现)与对照组(无胆道受侵表现)各6例。经超速离心法提取、Western Blot鉴定两组胆汁中的外泌体结构,采用Trizol法提取外泌体总RNA,再经miRNA表达谱芯片实验检测并寻找差异表达的miRNA。根据差异miRNA预测胆道侵犯靶基因,并进行通路富集分析。结果 观察组与对照组比较,共有74个差异表达的miRNAs,其中9个表达上调,65个表达下调(|Fold Change|>2)。经Pathway分析,靶基因主要富集在肿瘤发生、磷脂酰肌醇信号系统、PTEN等通路中(FDR<0.05)。经GO注释及富集分析,靶基因主要富集在基因抑制的正向调控、调控细胞分化等生物过程(FDR<0.05)。结论 建立肝泡型包虫病合并胆道侵犯胆汁中外泌体差异miRNA表达谱,可作为泡型包虫病侵及胆道的生物学标志物,初步阐释了肝泡型包虫病侵及胆道后差异miRNA对靶基因的调控机制。
【Abstract】:Objective To investigate the expression of exosome microRNAs (miRNAs) in bile of hepatic alveolar echinococcosis (HAE) patients with biliary tract invasion and the regulatory mechanism of differentially expressed miRNAs on target genes. Methods Bile samples were collected from 12 HAE patients who attended Qinghai University Affiliated Hospital from August 2017 to October 2018, with 6 patients in observation group (with the manifestation of biliary tract invasion) and 6 in control group (without the manifestation of biliary tract invasion). Ultracentrifugation extraction and Western blot were used to identify the structure of exosomes, the Trizol method was used to extract total RNA in exosomes, and miRNA expression profile microarray was used to identify differentially expressed miRNAs. The pathway enrichment analysis was performed to predict the target genes of biliary tract invasion based on differentially expressed miRNAs. Results A total of 74 differentially expressed miRNAs were identified between the observation group and the control group, among which 9 were upregulated and 65 were downregulated (|Fold Change|>2). The pathway analysis showed that the target genes were mainly enriched in the pathways for tumorigenesis, phosphatidylinositol signaling system, and PTEN (FDR<0.05). The GO annotation and enrichment analysis showed that the target genes were mainly enriched in the biological processes such as positive regulation of gene repression and regulation of cell differentiation (FDR<0.05). Conclusion The established expression profile of differentially expressed exosome miRNAs in bile of HAE patients with biliary tract invasion can be used as biomarkers for biliary tract invasion of HAE and preliminarily elaborate on the regulatory mechanism of differentially expressed miRNAs on target genes after HAE invades the biliary tract.
【关键字】:棘球蚴病,肝; 胆汁; 外泌体; 微RNAs; 计算生物学
【Key words】:echinococcosis,hepatic; bile; exosomes; microRNAs; computational biology
【引证本文】:WANG ZX, GOU P, YU WH, et al. Measurement and bioinformatics analysis of exosomes microRNAs in bile of hepatic alveolar echinococcosis patients with biliary tract invasion[J]. J Clin Hepatol, 2020, 36(9): 2045-2049. (in Chinese)
王志鑫, 苟平, 于文昊, 等. 肝泡型包虫病侵及胆道患者胆汁外泌体microRNA的检测及生物信息学分析[J]. 临床肝胆病杂志, 2020, 36(9): 2045-2049.

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