Differential Expression Profile Screening And Function Prediction Analysis Of CircRNA In Type 2 Diabetes Mellitus In Huadu District,Guangzhou

ObjectiveIn this study,circRNA microarray technology was used to explore the differential expression profile of circRNA in peripheral blood of type2 diabetes in Huadu District,Guangzhou,and to further verify the correlation between circRNA expression level and type 2 diabetes in the population,by analyzing the differentially expressed circRNA and correlation between biochemical indicators and type 2 diabetes,explore the possibility of differential expression of circRNA as a biomarker of type 2 diabetes.Finally predict the functional mechanism of differential expression of circRNA in the pathogenesis of type 2 diabetes,which provides a basis for the prevention and treatment of type 2 diabetes.MethodsThis study was based on a field residents over the age of 18 in Huadu District,Guangzhou,from January to October 2019.The individual matched case-control study design was used to include 107 patients with type 2 diabetes and 107 healthy controls selected with the same gender,age difference of 5 years as the matching condition.In the first stage,the peripheral blood of 4 patients with type 2 diabetes and 4 healthy controls were collected,and the circRNA differential expression profile was screened by circRNA microarray technology.In the second stage,whole blood samples and demographic as well as laboratory tests were collected from 103 patients with type 2 diabetes and 103 healthy controls.The four circRNAs screened in the circRNA differential expression profile were quantified by real-time fluorescence quantitative PCR.The ROC curve was used to evaluate the diagnostic value of the target circRNA in type 2diabetes.The multivariate conditional Logistic regression model was used to analyze the target circRNA and behavior factors for the impact of the risk of type 2 diabetes,the correlation between the target circRNA and biochemical indicators was explored using correlation analysis.GO analysis was used to predict the possible function of the target circRNA in the pathogenesis of diabetes,and KEGG pathway analysis was used to predict the signal pathway that the target circRNA may participate in the pathogenesis of diabetes.The database established by the software Epidata3.1 was used to cross-enter the questionnaire data,and the statistical analysis method was completed by the software SAS9.4 and R3.6.0.Results1.The circRNA microarray results showed that the circRNA differential expression profiles of the diabetic group and the healthy control group were initially established.A total of 86 circRNAs were found to be differentially expressed in diabetes.70 circRNAs were up-regulated and 16 circRNAs were down-regulated.2.The results of quantitative real-time PCR showed that the four significantly different circRNAs screened in the preliminary screening in the expanded population verification,only the expression of has?circ?0071106 between the two groups was statistically significant,and the expression of has?circ?0071106 in the diabetic group was up-regulated,compared with circRNA microarray results are consistent.In the multivariate conditional Logistic regression model,after controlling for mixed factors such as occupation and blood lipids,the risk of diabetes with a high expression level of has?circ?0071106 was 2.819 times the risk of disease with a low expression level(95%CI: 1.415 ? 5.616).Correlation analysis showed that there was a correlation between the expression level of has?circ?0071106 and blood glucose,low-density lipoprotein,and aspartate aminotransferase(P<0.05).The greater the blood glucose level or the greater the LDL value,the higher the expression level of has?circ?0071106,and the higher the aspartate aminotransferase concentration,the lower the expression level of has?circ?0071106.After adjusting other factors,the ROC curve analysisresults showed that the area under the curve has?circ?0071106 is 0.690,sensitivity of 62.1%,and specificity of 69.9%,suggesting that has?circ?0071106 can be considered as a biomarker for the diagnosis of type 2 diabetes.3.Functional prediction analysis results show that the five miRNAs combined with has?circ?0071106 are hsa-mi R-6830-3p,hsa-miR-4743-3p,hsa-miR-2682-3p,hsa-mi R-1206,hsa-mi R-29a-5p.GO functional analysis found that differentially expressed circRNAs are involved in biological processes such as protein binding or regulating gene transcription,and KEGG results predict that differentially expressed circRNAs are involved in MAPK signaling pathways that regulate insulin signal transduction.ConclusionThere is a differential expression profile of circRNA in the peripheral blood of type 2 diabetes and healthy people in a town of Huadu District,Guangzhou.has?circ?0071106 can be considered as a biomarker for the diagnosis of type 2 diabetes.It may be involved in the pathway of hsa-miR-29a-5p regulating diabetes,and may be involved in protein binding or biological processes regulating gene transcription,and may also be involved in the MAPK signaling pathways related to the onset of diabetes.