Elevated Circulating LINC-P21 Serves as a Diagnostic Biomarker of Type 2 Diabetes Mellitus and Regulates Pancreatic β-cell Function by Sponging miR-766-3p to Upregulate NR3C2

Abstract

Objective The purpose of this study was to evaluate the clinical value and biological function of long non-coding RNA (lncRNA) LINC-P21 in type 2 diabetes mellitus (T2DM), and explore the underlying mechanisms.

Methods The expression of LINC-P21 was estimated using quantitative real-time PCR. The functional role of LINC-P21 was explored by gain- and loss-of-function experiments. INS-1 cell proliferation was analyzed using a cell counting kit-8 (CCK-8)assay, and the glucose-stimulated insulin secretion was measured using an ELISA kit. The miRNAs that might be sponged by LINC-P21 were analyzed, and the subsequent target genes were predicted and assessed in INS-1 cells.

Results Serum expression of LINC-P21 was elevated in T2DM patients, which was correlated with fasting blood glucose levels and disease diagnosis. The glucose-stimulated insulin secretion and the proliferation of INS-1 cells were enhanced by LINC-P21 knockdown, but the overexpression of LINC-P21 led to opposite effects. miR-766-3p could be directly inhibited by LINC-P21 in INS-1 cells and reverse the effects of LINC-P21 on β-cell function. Additionally, NR3C2 was determined as a target of miR-766-3p, which could be positively regulated by LINC-P21 and had same effects with LINC-P21 on INS-1 cell proliferation and insulin secretion.

Conclusion All the data demonstrated that serum elevated LINC-P21 and decreased miR-766-3p serve as candidate diagnostic biomarkers in T2DM patients. LINC-P21 acts as a potential regulator in insulin secretion and proliferation of pancreatic β-cells through targeting miR-766-3p to upregulate NR3C2.

Key words

Type 2 diabetes mellitus - LINC-P21 - MicroRNA-766-3p - NR3C2 - Pancreatic β-cell - Proliferation

Publikationsverlauf

Eingereicht: 12. Juni 2020
Eingereicht: 13. August 2020

Angenommen: 24. August 2020

Artikel online veröffentlicht:
02. Oktober 2020

• References

• 1 Kao KT, Sabin MA. Type 2 diabetes mellitus in children and adolescents. Aust Fam Physician 2016; 45: 401-406
• 2 Shaw JE, Sicree RA, Zimmet PZ. Global estimates of the prevalence of diabetes for 2010 and 2030. Diabetes Res Clin Pract 2010; 87: 4-14 doi: 10.1016/j.diabres.2009.10.007
• 3 Gerber PA, Rutter GA. The role of oxidative stress and hypoxia in pancreatic beta-cell dysfunction in diabetes mellitus. Antioxid Redox Signal 2017; 26: 501-518 doi: 10.1089/ars.2016.6755
• 4 Ardestani A, Li S, Annamalai K. et al. Neratinib protects pancreatic beta cells in diabetes. Nat Commun 2019; 10: 5015
• 5 Robertson GL. Diabetes insipidus: Differential diagnosis and management. Best Pract Res Clin Endocrinol Metab 2016; 30: 205-218 doi: 10.1016/j.beem.2016.02.007
• 6 Roden M. Diabetes mellitus: Definition, classification and diagnosis. Wien Klin Wochenschr 2016; 128 Suppl 2: S37-S40
• 7 Jackson SL, Safo SE, Staimez LR. et al. Glucose challenge test screening for prediabetes and early diabetes. Diabet Med 2017; 34: 716-724
• 8 Prentki M, Nolan CJ. Islet beta cell failure in type 2 diabetes. J Clin Invest 2006; 116: 1802-1812 doi: 10.1172/JCI29103
• 9 Cnop M, Welsh N, Jonas JC. et al. Mechanisms of pancreatic beta-cell death in type 1 and type 2 diabetes: many differences, few similarities. Diabetes 2005; 54: S97-S107
• 10 Vishnoi A, Rani S. MiRNA Biogenesis and Regulation of Diseases: An Overview. Methods Mol Biol 2017; 1509: 1-10 doi: 10.1007/978-1-4939-6524-3_1
• 11 Shi X, Sun M, Liu H. et al. Long non-coding RNAs: A new frontier in the study of human diseases. Cancer Lett 2013; 339: 159-166
• 12 Pan X, Shen C, Huang Y. et al. Loss of SNHG4 attenuated spinal nerve ligation-triggered neuropathic pain through sponging miR-423-5p. Mediators Inflamm 2020; 2020: 2094948.
• 13 Guay C, Abdulkarim B, Tan JY. et al. Loss-of-function of the long non-coding RNA A830019P07Rik in mice does not affect insulin expression and secretion. Sci Rep 2020; 10: 6413
• 14 Arnes L, Akerman I, Balderes DA. et al. Betalinc1 encodes a long noncoding RNA that regulates islet beta-cell formation and function. Genes Dev 2016; 30: 502-507
• 15 You L, Wang N, Yin D. et al. Downregulation of Long Noncoding RNA Meg3 Affects Insulin Synthesis and Secretion in Mouse Pancreatic Beta Cells. J Cell Physiol 2016; 231: 852-862
• 16 Sathishkumar C, Prabu P, Mohan V. et al. Linking a role of lncRNAs (long non-coding RNAs) with insulin resistance, accelerated senescence, and inflammation in patients with type 2 diabetes. Hum Genomics 2018; 12: 41
• 17 Isin M, Uysaler E, Ozgur E. et al. Exosomal lncRNA-p21 levels may help to distinguish prostate cancer from benign disease. Front Genet 2015; 6: 168
• 18 Han W, Liu J. LncRNA-p21 inhibited the proliferation of osteosarcoma cells via the miR-130b/PTEN/AKT signaling pathway. Biomed Pharmacother 2018; 97: 911-918 doi: 10.1016/j.biopha.2017.11.014
• 19 Wang T, Liu J, Li S. et al. Effects of knockout of lincRNA-p21 on the proliferation, migration and invasion ability of HepG2 liver cancer cells. Oncol Lett 2019; 17: 5103-5107
• 20 Alberti KG, Zimmet PZ. Definition, diagnosis and classification of diabetes mellitus and its complications. Part 1: Diagnosis and classification of diabetes mellitus provisional report of a WHO consultation. Diabet Med 1998; 15: 539-553 doi: 10.1002/(SICI)1096-9136(199807)15:7<539::AID-DIA668>3.0.CO;2-S
• 21 Gong J, Liu C, Liu W. et al. LNCediting: a database for functional effects of RNA editing in lncRNAs. Nucleic Acids Res 2017; 45: D79-D84
• 22 Agarwal V, Bell GW, Nam JW. et al. Predicting effective microRNA target sites in mammalian mRNAs. Elife 2015; 4.
• 23 Sohrabifar N, Ghaderian SMH, Alipour Para S. et al. Variation in the expression level of MALAT1, MIAT and XIST lncRNAs in coronary artery disease patients with and without type 2 diabetes mellitus. Arch Physiol Biochem 2020; 1-8
• 24 Vijayan M, Reddy PH. Non-coding RNAs based molecular links in type 2 diabetes, ischemic stroke, and vascular dementia. J Alzheimers Dis 2020; 75: 353-383 doi: 10.3233/JAD-200070
• 25 Jiang N, Meng X, Mi H. et al. Circulating lncRNA XLOC_009167 serves as a diagnostic biomarker to predict lung cancer. Clin Chim Acta 2018; 486: 26-33
• 26 Li FP, Lin DQ, Gao LY. LncRNA TUG1 promotes proliferation of vascular smooth muscle cell and atherosclerosis through regulating miRNA-21/PTEN axis. Eur Rev Med Pharmacol Sci 2018; 22: 7439-7447 doi: 10.26355/eurrev_201811_16284
• 27 Zhang W, Zheng J, Hu X. et al. Dysregulated expression of long noncoding RNAs serves as diagnostic biomarkers of type 2 diabetes mellitus. Endocrine 2019; 65: 494-503
• 28 Liu SX, Zheng F, Xie KL. et al. Exercise Reduces Insulin Resistance in Type 2 Diabetes Mellitus via Mediating the lncRNA MALAT1/MicroRNA-382-3p/Resistin Axis. Mol Ther Nucleic Acids 2019; 18: 34-44
• 29 Song M, Zou L, Peng L. et al. LncRNA NONRATT021972 siRNA normalized the dysfunction of hepatic glucokinase through AKT signaling in T2DM rats. Endocr Res 2017; 42: 180-190
• 30 Tan J, Tong A, Xu Y. Pancreatic beta-cell function is inhibited by miR-3666 in type 2 diabetes mellitus by targeting adiponectin. Braz J Med Biol Res 2019; 52: e8344. doi: 10.1590/1414-431X20198344
• 31 Jiang H, Liu Y, Qian Y. et al. CHL1 promotes insulin secretion and negatively regulates the proliferation of pancreatic beta cells. Biochem Biophys Res Commun 2020; 525: 1095-1102
• 32 Paraskevopoulou MD, Hatzigeorgiou AG. Analyzing MiRNA-LncRNA Interactions. Methods Mol Biol 2016; 1402: 271-286 doi: 10.1007/978-1-4939-3378-5_21
• 33 Yang ZM, Chen LH, Hong M. et al. Serum microRNA profiling and bioinformatics analysis of patients with type 2 diabetes mellitus in a Chinese population. Mol Med Rep 2017; 15: 2143-2153
• 34 You Y, Que K, Zhou Y. et al. MicroRNA-766-3p Inhibits Tumour Progression by Targeting Wnt3a in Hepatocellular Carcinoma. Mol Cells 2018; 41: 830-841
• 35 Yang C, Ma X, Guan G. et al. MicroRNA-766 promotes cancer progression by targeting NR3C2 in hepatocellular carcinoma. FASEB J 2019; 33: 1456-1467
• 36 Gragnoli C. Hypothesis of the neuroendocrine cortisol pathway gene role in the comorbidity of depression, type 2 diabetes, and metabolic syndrome. Appl Clin Genet 2014; 7: 43-53 doi: 10.2147/TACG.S39993