A preliminary research on the relationship of hsa_circ_0031739 and type 2 diabetes mellitus / 中华检验医学杂志

ABSTRACT

Objective In order to establish a basis for exploring diagnostic biomarkers of type 2 diabetes mellitus (T2DM), our research screened differentially expressed circRNAs in high glucose treated human umbilical vein endothelial cells ( HUVECs) and validated these circRNAs in both cell models and peripheral blood samples of T 2DM patients.Methods The research used HUVECs as experimental model . The control group (n=3) and high glucose group (n=3) were set up and treated under normal condition (5.5 mmol/L) and high glucose condition (30.0 mmol/L), respectively.The research utilized high-throughput sequencing technology to preliminarily screen differentially expressed circRNAs .Differentially expressed circular RNAs were validated in the endothelial cell model by PCR and real -time quantitative PCR ( q-PCR) techniques.Subsequently , in the peripheral blood samples of T 2DM patients ( n=32) and control individuals ( n=28 ), the differentially expressed circRNAs were further validated . Finally, t-Test, correlation analysis and ROC curve were used to analyze the experimental results .Results Total 1087 differentially expressed circular RNAs were screened by high-throughput sequencing;and of these , 554 were up-regulated, 533 were down-regulated.Six increased circRNAs were selected and validated in HUVEC model;and their PCR and q-PCR results matched with sequencing results .Further validation was conducted in peripheral blood samples for three most up-regulated circRNAs.This study found that the differential expression of hsa_circ_0031739 in peripheral blood samples was statistically significant ( 0.015 ±0.0025 vs.0.006 ±0.0013,P=0.0059 <0.05).The area under the ROC curve (AUC) was 0.730 and the expression level of hsa_circ_0031739 was positively correlated with blood glucose ( GLU ) and glycated hemoglobin ( GHb ) ( r=0.317, P=0.0137 <0.05;r=0.348, P=0.0064 <0.05 ) .Conclusion HUVECs were commonly utilized as cell models in the study of T 2DM, and differentially expressed circular RNA profiles existed afterhigh glucose treatment .The differential expression of hsa_circ_0031739 was significant in both high glucose treated HUVECs and peripheral blood samples of T 2DM patients, and the expression level of hsa_circ_0031739 was correlated with GLU and GHb , which has certain diagnostic significance.Therefore, hsa_circ_0031739 may become a new diagnostic biomarker for T2DM and be helpful for the comprehensive diagnosis and mechanism research of T 2DM.