A single nucleotide polymorphism located in microRNA-499a causes loss of function resulting in increased expression of osbpl1a and reduced serum HDL level

Abstract

Atherosclerosis is the main pathological process that induces CVD (cardiovascular diseases), and the objective of our study was explore whether miR‑499a rs3746444 polymorphism was associated with the HDL level, one of the risk factors of atherosclerosis. Online public miRNA database was utilized to predict the miR‑499a target, and luciferase assay was conducted to confirm that miR‑499a targeted osbpl1a, then western blot analysis and real-time PCR were performed to verify miRNA-mRNA regulatory relationship between miR‑499a and osbpl1a. Based on results of bioinformatics algorithms, osbpl1a was predicted as a candidate target gene of miR‑499a, luciferase reporter was generated, and it was found that the luciferase activity of cells was substantial downregulated following co-transfection with wild osbpl1a 3'UTR and miR‑499a compared to that in scramble control, while the inhibitory effect of miR‑499a was abolished after transfection of mutant osbpl1a 3'UTR. Then, miRNA-mRNA regulatory relationship between miR‑499a and osbpl1a was detected, a concentration-dependent effect of miR‑499a on the miR‑499a expression was observed, and both osbpl1a mRNA and protein levels of cells transfected with miR‑449a (30 and 60 nM) or osbpl1a siRNA were markedly reduced, while notably improved subsequent to transfect with anti‑miR‑449a (30 and 60 nM) in comparison with NC groups, moreover, the inhibitory effect among 30 or 60 nM miR‑499a, osbpl1a siRNA was similar, the improved effect of 30 or 60 nM anti‑miR‑499a showed no significant change. The influence of rs3746444 A allele on expression level of miR‑499a represented a recessive pattern in high-grade group with a higher level of miR‑499a in AA group, and HDL level in AA group was significantly reduced related to those in AG and GG groups. This study validated that rs3746444 polymorphism influenced the expression of miR‑499a, its target gene, osbpl1a, and thereby associated with the HDL level, which makes it a potential factor involved in the mechanism of atherosclerosis.

miRNA Polymorphisms and Risk of Cardio-Cerebrovascular Diseases: A Systematic Review and Meta-Analysis

“…Theoretically, rs3746444 may increase the risk of CCD by either altering miR-499a-5p/3p (or possibly miR-499b-5p/3p) expression or influencing the repertoire of target genes of miR-499a-3p (or possibly miR-499b-5p), or a combination of both scenarios. A recent study provided support for the first scenario [56], demonstrating that rs3746444 can modulate the expression of miR-499a. This study showed that the rs3746444-A allele may be correlated with higher serum levels of miR-499a-5p and lower HDL in a recessive manner (i.e., AA vs. AG+GG).…”

“…Although there is no general consensus about the treating of HWE-deviated association studies, it has been recommended that the meta-analysis may benefit from including such studies [55]. However, sensitivity analysis should be performed to assess the possible influences of such studies on the conclusion of the meta-analysis [18,56,57,58]. Therefore, this study included HWE-deviated studies in the meta-analyses while assessing the potential influence of the HWE departure on the overall and subgroup analyses.…”

Associations of the miRNA-146a rs2910164 and the miRNA-499a rs3746444 Polymorphisms With Plasma Lipid Levels: A Meta-Analysis

“…Moreover, macrophages transfected with miRNA-146a mimics reduced the intracellular LDL-C and oxLDL accumulation (Yang et al, 2011 ). However, transfection of HepG2 cells with the miRNA-499a inhibitor significantly increased the intracellular HDL-C levels (Chen et al, 2017 ). Together, it indicated that the expression levels of miRNA-146a and miRNA-499a may be closely linked to lipid metabolism.…”

“…The rs3746444 polymorphism is a function variant of miRNA-499a , formed by a nucleotide substitution from adenine (A) to guanine (G). Evidence showed that the G allele of the rs3746444 polymorphism largely decreased the miRNA-499a protein levels (Chen et al, 2017 ). Therefore, the rs2910164 and rs3746444 polymorphisms may affect lipid levels by altering the expression levels of miRNA-146a and miRNA-499a .…”

Association analysis of miR‐499 rs3746444 gene polymorphism with periodontitis

“…Aberrant expression of miRNAs may tilt the tissue homeostasis leading to the development of many chronic inflammatory diseases. The polymorphisms of miR‐499 rs3746444 influenced the expression of miR‐499 (Chen et al, 2017). This SNP has been associated with many diseases like papillary thyroid carcinomas, prostate cancer and rheumatoid arthritis (Fan, Chen, & Wu, 2013; Hashemi et al., 2013).…”