Role and Significance of Circulating Biomarkers: miRNA and E2F1 mRNA Expression and Their Association with Type-2 Diabetic Complications

Abstract

Background. Type 2 diabetes mellitus (T2DM) has emerged as an epidemic affecting more than four hundred million people throughout the world. It is a multifactorial disease with range of environmental and genetic factors responsible for its prevalence. In search of novel biomarkers for recording progress of various metabolic diseases, small noncoding RNA in general and microRNAs (miRNAs) in particular have emerged as the most promising biomarkers for diagnosing variety of diseases including diabetes. An increasing number of studies have been published, reporting the quantification of miRNAs in blood of subjects with diabetes and mostly aimed at identifying miRNA modulation in chronic diabetic complications. Due to its association with immune system homeostasis and potential capability to predict diabetes development, the profile of circulating miRNAs may also provide useful information about diabetes pathogenic mechanisms. Thus, the present study aimed to understand the role and expression of microRNA330 and E2F1 mRNA expression in patients with T2DM. Methodology. The present study includes a total of 200 individuals: 100 “individuals with T2DM referred to as “cases” and 100 healthy individuals referred to as “controls”. Extracted RNA was used to synthesise the cDNA for microRNA-330 and E2F1 mRNA expression. Taqman assay method has been used to analyse the microRNA-330 expression in the cases and controls and SYBR green dye was used to study the E2F1 mRNA expression. Results. Statistically significant difference was observed in all the selected 5 biochemical parameters among T2DM cases and healthy controls. Risk factors like hypertension were observed to be significantly associated with reduced HDL (p=0.01), increased TG (p=0.0008), and cholesterol (p<0.0001) in hypertensive T2DM cases as compared to nonhypertensive T2DM cases. Obese patients showed significant increase in TG (p=0.01) and cholesterol (p<0.0001) as compared to nonobese patients. Similarly, increased TG (p=0.001) and cholesterol (p<0.0001) was observed in the case of alcoholic patients as compared to nonalcoholic patients. Also, patients with smoking habit showed increased TG (p=0.009p = 0.009), cholesterol (p<0.0001), and VLDL (p=0.01) as compared to nonsmokers and differences among them was found to be statistically significant. Besides this, significant impact of risk factors like hypertension, obesity, alcoholism, and smoking were observed on microRNA-330 expression and E2F1 mRNA expression. A 7.72-fold increased microRNA-330 and 0.05-fold decreased E2F1 mRNA expression was observed among T2DM cases as compared to healthy controls. Increased expression of microRNA-330 was observed in hypertensive cases (9.61-fold, p<0.0001), obese cases (9.33-fold, p=0.0008, alcoholic cases (9.07-fold, p<0.0001), and smoking cases (8.41-fold, p=0.01) as compared to nonhypertensive, nonobese nonalcoholic, and nonsmoking cases, and differences among them were found to be significant. Decreased expression of E2F1 mRNA expression was observed in patients with alcoholism (0.03-fold, p=0.002) and smoking (0.03fold, p<0.0001) while patients who were nonalcoholic and nonsmokers showed 0.07-fold increase in expression, and differences among them were found to be statistically significant. Conclusion. The present study demonstrated that increased level of microRNA-330 and decreased level of E2F1 mRNA expression were found to be associated with pathogenesis of T2DM patients. Risk factors such as hypertension, obesity, alcoholism, and smoking may be were found to be associated with microRNA-330 and E2F1 mRNA expressions, and it can prove a reliable biomarker for T2DM disease progression could be linked to chronic diabetic complications.

Identification of novel prognostic targets in acute kidney injury using bioinformatics and next generation sequencing data analysis

“…IGF2 [494], IRF7 [495], E2F1 [496], TEAD4 [497], KCNH2 [498], E2F2 [499], SNHG7 [500], FLI1 [501], CYP3A5 [502], HMGCS2 [503], GOT1 [504], PPARGC1A [505], GC (GC vitamin D binding protein) [506], VNN1 [507], NOX4 [508], SLC2A1 [509], BPGM (bisphosphoglyceratemutase) [164], NR4A3 [354], PFKFB2 [510], CDH2 [511], F11 [512], AQP2 [513], CLDN2 [514], EGF (epidermal growth factor) [515], ANGPT1 [516], KNG1 [517], SERPINA5 [518], HRG (histidine rich glycoprotein) [519], KL (klotho) [520], DEFB1 [521], ACE2 [522], AQP3 [523], CADM1 [188], DPP4 [524], STC1 [525], REN (renin) [526], TRPM6 [527], MSR1 [528], CCR1 [529], TNFRSF11B [530], FZD5 [531], ERBB4 [216], F8 [532], VCAM1 [533], PTGER3 [534] and ALB (albumin) [535] have been shown to influence the genetic risk of sepsis. IGF2 [536], IRF7 [100], PRKCB (protein kinase C beta) [101], CCL5 [537], EEF1A2 [538], ACTN3 [264], FCN1 [539], BRSK2 [540], MNX1 [541], AMH (anti-Mullerian hormone) [542], E2F1 [543], HAP1 [544], PF4 [545], AGER (advanced glycosylation end-product specific receptor) [546], E2F2 [547], TYMP (thymidine phosphorylase) [548], PPP1CC [549], NR2E1 [550], GREM1 [436], GRIN1 [551], WNT3A [552], COMP (cartilage oligomeric matrix protein) [553], BHMT (betaine--homocysteine S-methyltransferase) [554], ANGPTL3 [555], PCK1 [556], KMO (kynurenine 3-monooxygenase) [557], HSD11B2 […”

“…The identified hub gens, TFs and miRNAs might be involved in the pathological process of AKI. PPP1CC [549], E2F1 [543], EEF1A2 [538], VCAM1 [626], CDH1 [622], AGTR1 [79], SNCA (synuclein alpha) [574], hsa-mir-221 [780], YY1 [781], STAT3 [782] and PDX1 [783] are closely involved with diabetes mellitus. E2F1 [110], BMI1 [95], EEF1A2 [104], ACTA1 [128], VCAM1 [220], AGTR1 [58], hsa-mir-221 [784], MEF2A [785], FOXC1 [786], YY1 [787], STAT3 [788] and BRCA1 [789] were significantly associated with cardiovascular disorders.…”

Bioinformatics Analysis of Next Generation Sequencing Data Identifies Molecular Biomarkers Associated With Type 2 Diabetes Mellitus

“…The roles of hub genes, miRNA and TF in the pathogenesis of T2DM are discussed. Hsa-mir-142-5p , 235 hsa-mir-1291, 236 NOTCH1 , 237 PPARD (peroxisome proliferator-activated receptor delta), 238 HIF1A , 239 RUNX2 , 240 and E2F1 241 levels are correlated with disease severity in patients with T2DM. Previous studies have demonstrated that hsa-mir-216b-5p 242 and hsa-mir-200a-3p 243 appears to be expressed in Type 1 diabetes.…”

Analysis of key genes and pathways associated with the pathogenesis of Type 2 diabetes mellitus

“…The roles of hub genes, miRNA and TF in the pathogenesis of T2DM are discussed. Hsa-mir-142-5p [231], hsa-mir-1291 [232], NOTCH1 [233], PPARD (peroxisome proliferator-activated receptor delta) [234], HIF1A [235], RUNX2 [236] and E2F1 [237] levels are correlated with disease severity in patients with T2DM. Previous studies have demonstrated that hsa-mir-216b-5p [238] and hsa-mir-200a-3p [239] appears to be expressed in type 1 diabetes, but these genes might be novel target for T2DM.…”