Differential Expression Validation And Functional Prediction Of Essential Hypertension-related CircRNAs

Objective: This paper aims to verify the differential expression of differentially expressed circ RNAs in human screened by microarray technology by using q RT-PCR technology,and predict its possible mechanism of action by combining bioinformatics technology,so as to provide ideas for further exploration of circ RNA as a biomarker for diagnosis of essential hypertension(EH)and therapeutic target.Methods: According to the unified inclusion and exclusion criteria and the principle of frequency matching,200 local Han nationality residents in Ningbo city were selected in this study,including 100 patients with EH and 100 healthy controls.Baseline data were investigated and collected by uniformly trained investigators.Blood samples were collected and total RNA was extracted and transcribed into c DNA.The relative expression levels of candidate differentially expressed circ RNAs(hsa＿circ＿0093587,hsa＿circ＿0017170,hsa＿circ＿0037897 and hsacirc RNA9102-5)in the blood samples of the study subjects were detected by q RT-PCR.Single factor analysis was used to compare the relative expression of target circ RNAs between the two groups and evaluate its efficacy as a diagnostic biomarker using ROC curve.Circ RNAs-related mi RNAs and target genes were predicted,and the related functions and enrichment pathways of target genes were analyzed by bioinformatics techniques.Circ RNA-mi RNA pathway model was constructed by combining the expression level of target mi RNAs.Results: 1.The four candidate differentially expressed circ RNAs from 287 differentially expressed circ RNAs screened by microarray in the early stage were verified by q RT-PCR.The results showed that the expression levels of hsa＿circ＿0017170,hsa＿circ＿0037897 and hsacirc RNA9102-5 were increased in the EH group(low △Ct value),which was consistent with the microarray results.There was no significant difference in hsa＿circ＿0093587 expression between the EH group and the healthy control group.In the age group,only hsa＿circ＿0037897 had a higher relative expression in the age≥60 group.There was no significant difference between the two groups in terms of gender,smoking,drinking and BMI.2.Establish ROC curve to evaluate the diagnostic value of each circ RNAs and their combined effects on EH.Except that the relative expression of hsa＿circ＿0093587 has no diagnostic value for EH,the other three circ RNAs have certain diagnostic value.Hsa＿circ＿0017170 has the highest diagnostic value of AUC of 0.652(95%CI: 0.5766-0.727),with sensitivity and specificity of 0.620 and 0.640,respectively.When three circ RNAs were applied jointly,the AUC reached 0.709.3.It was found by the prediction software that circ RNAs with different expressions verified could combine with many different types of mi RNAs,including mi RNAs related to EH,such as hsa-mi R-145-5p and hsa-mi R-150-5p,etc.Through GO and KEGG analysis,it was found that mi R-145-5p regulates the expression of target genes and participates in the transport of calcium ions and the Wnt signaling pathway and mi R-150-5p is involved in steroid-hormone-mediated signaling pathways which may be associated with EH.Conclusion: This study verified that hsa＿circ＿0017170,hsa＿circ＿0037897 and hsacircrna9102-5 were up-regulated in EH group and their high expression in the human body were independent risk factors for EH.The results of experimental results and bioinformatics techniques suggested the possible mechanism of circ RNAs regulating the occurrence and development of EH.This study provides a direction for circ RNAs to be used as diagnostic markers and gene targets for EH.