Study On The Bile Exosomes Of Hepatic Alveolar Echinococcosis With Biliary Tract Invasion

Objective: To detect the expression characteristics of exosomes miRNAs in bile of hepatic alveolar echinococcosis(HAE)with biliary tract invasion,and to search for biomarkers of biliary tract invasion,laying a foundation for the advanced study on the mechanism of biliary tract invasion.Methods: We collected a total of 12 patients' bile samples diagnosed as HAE from August 2017 to October 2018 in the Affiliated Hospital of Qinghai University,including 6 patients in the observation group(biliary tract invasion group)and 6 patients in the control group(not biliary tract invasion group).Total RNA in exosomes was extracted by TRIZOL method after extraction through ultracentrifugation,Western Blotting detection and transmission electron microscopy with the purpose of identification of exosomes structure in bile.Then miRNAs with differential expression were searched by miRNAs expression profile chip experiment.Pathway enrichment analysis was performed to predict target genes of biliary tract invasion based on differential miRNAs.Results: The concentrations of bile exosomes extracted by the ultracentrifugation method in each group could meet the requirements of Western Blotting detection and chip sampling of miRNAs expression profile.After the chip experiment,a total of 74 differentially expressed miRNAs between the observation group and the control group,of which 9 were up-regulated and 65 were down-regulated.45 target genes of biliary tract invasion were predicted by differential miRNAs,such as AFF3,STK4,PIK3R1,etc.Through Pathway analysis,the target genes were mainly enriched in Pathways in cancer,Phosphatidylinositol signaling system,PTEN and other pathways.By GO annotation and enrichment analysis,the target genes were mainly enriched in biological processes,such as positive regulation of gene repression,regulation of cell differentiation.Conclusions: Established the expression profile of exosomes differential miRNAs in bile of HAE with biliary tract invasion,and these differentially expressed miRNAs can be used as candidate biomarkers for biliary tract invasion.The regulatory network of differential miRNAs on target genes was preliminarily explored,laying a foundation for the study on the mechanism of biliary tract invasion.