Analysis Of MiRNAs Specific Expression And Prediction Of Target Gene Signal Pathway In Hypertrophic Cardiomyopathy

Objective1 To explore the expression profile of miRNAs in the peripheral blood of patients with hypertrophic cardiomyopathy and to screen out the miRNAs specifically expressed in the peripheral blood of patients with hypertrophic cardiomyopathy;2 To predict the target gene and signal pathway of miRNAs with significant differential expression and to explore the possible pathogenesis of hypertrophic cardiomyopathy.Methods1 A total of 3 patients with hypertrophic cardiomyopathy diagnosed by cardiac ultrasound or imaging from March 2018 to October 2018 in the inpatient and outpatient departments of our hospital were included as HCM group and 3 healthy people who underwent examination in the physical examination center of our hospital were included as healthy control group.The clinical data collection was completed,and the peripheral blood was extracted and centrifuged to extract plasma for storage.The miRNAs expression profile was obtained by miRNA chip detection technology,and miRNAs with significant differential expression were screened by screening conditions and combined with literature reports,and the screened miRNAs were verified by RT-PCR.2 A total of 60 patients with hypertrophic cardiomyopathy diagnosed by cardiac ultrasound or imaging from March 2018 to November 2019 in the inpatient and outpatient departments of our hospital were included as HCM group and a total of 60 healthy people who underwent examination in the physical examination center of our hospital were included as healthy control group.The clinical data collection was completed,and the peripheral blood was extracted and centrifuged to extract plasma for storage.Based on the results of the above three cases,several miRNAs with significant differences were screened and verified,and RT-PCR verification with expanded sample size was carried out.ROC curve was drawn for miRNAs with significant differential expression.3 Target genes of miRNAs with significant differential expression were predicted through miRanda,Target Scan,DIANA and Pictar databases.VENNY online database was used to get the intersection of the target genes predicted by any two databases to obtain common target genes.The Mate Scape online database was used for target gene GO enrichment analysis and KEGG or Reactome Gene Sets pathway analysis.Results1 A total of 952 miRNAs were obtained by miRNA chip detection technology.Based on the screening criteria of P value<0.05 and Fold Change>2 times,919 miRNAs with no significant difference and 33 miRNAs with differential expression were screened out.Combined with relevant literatures,10 miRNAs with significant differential expression were screened out(miR-2355-5p,let-7g-3p,miR-103a-3p,miR-122-5p,miR-146b-5p,miR-200c-5p,miR-377-3p,miR-675-3p,miR-26a-1-3p,miR-208b-3p).After RT-PCR verification,the results showed that the expression trends of 8 miRNAs were consistent with the detection results of miRNAs chip,among which the up-regulated miRNAs were let-7g-3p,miR-103a-3p,miR-122-5p,miR-146b-5p,miR-26a-1-3p,miR-208b-3p,and the down-regulated miRNAs were hsa-miR-675-3p,hsa-miR-377-3p.The other two were miR-2355-5p and miR-200c-5p,which were contrary to the chip detection results and showed downward regulation.2 Based on the 8 miRNAs with differential expression found in the previous research and combined with literature,2 miRNAs were screened out,namely miR-208b-3p and miR-122-5p respectively.Compared with the healthy control group(60 cases),the HCM group(60 cases)showed significant difference in the expression level of plasma miRNA-122-5p(P value<0.05),showed no significant difference in plasma miR-208b-3p(P value> 0.05),and the area under miRNA-122-5p ROC curve was0.865.3 Target genes of differentially expressed miR-122-5p were predicted through miRanda,Target Scan,DIANA and Pictar databases,and a total of 326 target genes were obtained.The Mate Scape online database was used for enrichment analysis of target genes.The results showed that GO functions related to miR-122-5p were mainly enriched in cell adhesion regulation,skeletal system development,cell connective tissue,aging,response to braking stress,axon protein transport,negative regulation of cell differentiation,negative regulation of microtubule polymerization,regulation of sodium ion transporter activity,establishment of mitochondrial membrane protein localization,regulation of interstitial cell apoptosis process,post-embryonic development,etc.The pathway analysis found that miR-122-5p regulated target genes were mainly involved in acetylation regulating TP53 activity,antigen processing and presentation,metabolism of carbohydrates,extra-nuclear estrogen signaling,glycogen storage diseases and other pathways.Conclusions1 Patients with hypertrophic cardiomyopathy have a specific miRNAs expression profile,and the expression of miR-122-5p in plasma is significantly increased,which is expected to be a diagnostic molecular marker for hypertrophic cardiomyopathy.2 miR-122-5p may regulate TP53 activity through activation and acetylation of its target gene to participate in the occurrence of myocardial hypertrophy.