A preliminary research on the relationship of hsa_circ_0031739 and type 2 diabetes mellitus

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 T2DM 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 T2DM 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 1 087 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±0.002 5 vs. 0.006±0.001 3, P=0.005 9<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=0.31 7, P=0.013 7<0.05; r=0.348, P=0.006 4<0.05). Conclusion HUVECs were commonly utilized as cell models in the study of T2DM, 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 T2DM 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 T2DM.(Chin J Lab Med, 2018, 41: 863-869) Key words: Diabetes mellitus, type 2; RNA; Biomarkers; High-throughput nucleotide sequencing