Screening And Identification Of Serum-specific MicroRNAs For Alveolar Echinococcosis In Southern Of Qinghai Tibetan Region

ObjectiveMicroRNA microarray assay was used analysis the differentially expressed microRNAs in serum of alveolar echinococcosis(AE)in southern of Qinghai Tibetan areas.The differentially expressed serum microRNAs which related with AE were validated using quantitative Real-Time PCR(qRT-PCR)with large sample analysis.This disease-related microRNAs were screened as new biomarkers for AE.Methods1.Ten serum samples were collected from patients with AE,and ten serum samples were collected from matched healthy people.Total RNA was extracted from serum,and microRNA microarray assay was used for testing the serum microRNAs expression profiles.Analysis and compared the expression amount of microRNAs in the serum of patients with AE and healthy people,different expression microRNAs related with AE were found and screened out.2.Fifty serum samples were collected from patients with AE,and fifty serum samples were collected from matched healthy people.Combining with the results of microRNA microarray assay,the differentially expressed serum microRNAs were selected and validated using qRT-PCR.The relative expression of differentially expressed serum microRNAs were calculated,and compared the qRT-PCR results and microRNA microarray assay results,screened out the serum-specific miRNAs of AE.3.Bioinformatics software was used to predict the target genes of serum-specific microRNAs of AE,and analysis the target genes' GO function and KEGG pathway.Results1.The microRNA microarray assay results revealed that A total of 2615 miRNAs were detected in the AE group and control group.Among them,5 miRNAs were only expressed in the control group,they were has-miR-6891-5p,has-miR-4274,has-miR-3191-5p,has-miR-6750-3p,has-miR-6892-3p.And 34 miRNAs were only expressed in the AE group,they were has-miR-19b-3p,has-miR-5701,has-miR-4262,has-miR-6890-3p,has-miR-3202,has-miR-5698,has-miR-6769a-3p,has-miR-3074-3p,has-miR-6892-5p,has-miR-6769a-5p,has-miR-302 e,has-miR-6751-3p,has-miR-1468-3p,has-miR-8065,has-miR-8053,has-miR-200b-5p,has-miR-135b-5p,has-miR-4468,h as-miR-302b-3p,has-miR-301b-3p,has-miR-1910-3p,has-miR-30e-5p,has-miR-6870-3p,has-miR-133a-5p,has-miR-6882-5p,has-miR-1323,has-miR-1205,has-miR-6880-5p,has-miR-4665-3p,has-miR-8062,has-miR-7106-3p,has-miR-6893-5p,has-miR-320 c,has-miR-6871-5p.2.Compared the miRNA expression amount in AE patients and healthy people,there were 74 differentially expressed miRNAs(|log2(Fold change)| ?1,p-value 0.05),and among them,39 miRNAs were up-regulated and 35 miRNAs were down-regulated.3.qRT-PCR results showed that the relative expression of miRNA-4725-3p in serum of patients with AE was 1.26 times higher than that of healthy people(the difference was statistically significant,P<0.05).4.It was predicted that the target genes regulated by has-miRNA-4725-3p were mainly involved in PI3K-AKT signaling pathway,RAS signaling pathway,MAPK signaling pathway,Calcium signaling pathway,etc.ConclusionThe expression of miRNAs in the serum of patients with AE were different from that of healthy people.miRNA-4725-3p might be a new biomarker for AE.Target genes regulated by has-miRNA-4725-3p are mainly involved in PI3K-AKT signaling,RAS signaling,MAPK signaling and calcium signaling.However,the relationship between these target genes and AE needs to be further verified.