The Significance Of Plasma Exosomal MiRNA In The Diagnosis And Progression Of Hepatocellular Carcinoma

Background Hepatocellular carcinoma(HCC)accounts for more than 95% of primary liver cancer.Its pathogenesis is characterized by many factors,rapid progression,and no obvious symptoms in early stage.Therefore,most of the HCC has been detected and diagnosed in the middle and advanced stage.The clinical characteristics of "high morbidity,high fatality rate and low five-year survival rate" of HCC make it become one of the problems to be solved urgently in medicine and life science.In terms of clinical diagnosis and treatment,in addition to exploring the mechanism of HCC progression,it is also crucial to find stable and reliable diagnostic biomarkers and improve the diagnostic rate of HCC.Exosomes are intracavinal vesicles that bud from the inner membrane of cells.They contain many small molecule contents including non-coding RNA(nc RNA),which are transported to recipient cells by exosomes and play a role in information communication between cells.MicroRNAs(miRNAs),one of the nc RNAs,regulate the expression of target genes by complementary binding with intracellular target genes,and play an important role in biological development and disease development.As the medium of information communication between cells,miRNA can be selectively sorted into exosomes,and the miRNA content in exosomes may be different from that in mother cells.In addition,miRNAs are encapsulated in exosomes and can effectively prevent their degradation by RNA enzymes,so that they can be transferred to distant target cells and play their roles.Based on this characteristic,plasma exosome miRNAs derived from HCC cells are more reliable than plasma miRNAs as diagnostic markers.MethodsIn this study,140 volunteers(HCC patients and healthy control volunteers)were recruited to extract exosomes from plasma samples.High throughput sequencing was used to detect miRNA expression in 9 samples(3 HCC before surgery,3 HCC after surgery,and 3 healthy volunteers).Bioinformatics analysis and q PCR were used to screen candidate miRNAs.Logistic regression model was constructed using training queues,and then independent queue validation was used.The diagnostic accuracy was evaluated by the AUC of the ROC curve.In addition,bioinformatics analysis was used to screen exosomal miRNAs associated with HCC progression,and the correlation between candidate miRNAs and HCC progression was verified by q PCR.ResultsWe identified four groups of exosomal miRNAs(miR-212-5p,miR-519b-3p,miR-1248,and miR-1250-5p),which provided high diagnostic accuracy for HCC(AUC of training and validation datasets were 0.8450 and 0.8447,respectively).Meanwhile,Meanwhile,we found that the diagnostic efficacy AUC of the exosomal miRNA diagnostic panel combined with AFP for HCC was 0.9609.In addition,q PCR verified the exosomal miRNA associated with HCC progression based on sequencing analysis,and the expression of miR-1248 was significantly decreased in plasma exosomes of HCC patients(t=2.849,P < 0.05).Bioinformatics analysis showed that exosomal miR-1248 could inhibit the progression of HCC by regulating extracellular matrix receptor interactions and mediating metabolic reprogramming in recipient cells.ConclusionPlasma exosomal miRNA diagnostic panels(miR-212-5p,miR-519b-3p,miR-1248,and miR-1250-5p)can be used for the diagnosis of HCC.AFP combined with plasma exosomal miRNAs significantly improved the diagnostic accuracy of HCC.Plasma exosomal miR-1248 inhibited the progression of HCC.