microRNA-185 modulates low density lipoprotein receptor expression as a key posttranscriptional regulator

• miR-185 regulates LDLR expression by direct binding to LDLR 3′-UTR in HepG2 cells.
• KSRP, one of the LDLR mRNA-destabilizing proteins, is a direct target of miR-185.
• miR-185 mediates an indirect regulation of LDLR through KSRP.
• miR-185 inhibitor increases hepatic LDLR expression in vivo.
• miR-185 inhibitor reduced atherosclerosis progression in vivo.

ObjectiveLow-density lipoprotein receptor (LDLR) mediates endocytosis of LDL particles and is important in maintaining plasma cholesterol levels, thus its expression is under extensive regulation at multiple levels, including transcriptional and posttranscriptional regulation by transcription factors (TFs) and RNA-binding proteins (RBPs). Here, we identified microRNA-185 (miR-185) as a novel direct posttranscriptional regulator of LDLR and an indirect LDLR modulator through KSRP in hepatic cells.Methods and resultsUsing quantitative real-time PCR (qPCR), we detected the effect of predicted LDLR-targeting miRNAs and found that overexpression of miR-185 repressed LDLR expression and LDL uptake in HepG2 cells by 62.4 ± 6.0% (p = 7.0 × 10−5) and 32.5 ± 6.0% (p = 7.7 × 10−4) respectively, through directly targeting LDLR 3′UTR. Unexpectedly, the antisense inhibitor of miR-185 had similar repression effect on LDLR although it reduced the association of endogenous miR-185 with LDLR mRNA. Further experiments revealed that KH-type splicing regulatory protein (KSRP), one of the LDLR-destabilizing RBPs, is also a target of miR-185. KSRP silencing reversed the repression effects of miR-185-inhibitor on LDLR. Thus miR-185 regulates LDLR expression not only through directly targeting but also by a RBP-involved indirect pathway. Finally, the in vivo results showed that miR-185-inhibitor upregulated hepatic LDLR expression and correlated with a decrease in plasma cholesterol level and arterial plaque area in ApoE KO mice.ConclusionsThese findings reveal that miR-185 controls cholesterol homeostasis as a key posttranscriptional LDLR modulator in hepatic cells, providing novel insight into the regulatory mechanism for LDLR expression and the anti-atherosclerosis effect of miR-185-inhibitor.

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