H19 knockdown suppresses proliferation and induces apoptosis by regulating miR-148b/WNT/β-catenin in ox-LDL -stimulated vascular smooth muscle cells

Abstract

BackgroundLong non-coding RNAs (lncRNAs) have been identified as critical regulators in the development of atherosclerosis (AS). Here, we focused on discussing roles and molecular mechanisms of lncRNA H19 in vascular smooth muscle cells (VSMCs) progression.MethodsRT-qPCR assay was used to detect the expression patterns of H19 and miR-148b in clinical samples and cells. Cell proliferative ability was evaluated by CCK-8 and colony formation assays. Cell apoptotic capacity was assessed by apoptotic cell percentage and the caspase-3 activity. Bioinformatics analysis, luciferase and RNA immunoprecipitation (RIP) assays were employed to demonstrate cell percentage and the relationship among H19, miR-148b and wnt family member 1 (WNT1). Western blot assay was performed to determine expressions of proliferating cell nuclear antigen (PCNA), ki-67, Bax, Bcl-2, WNT1, β-catenin, C-myc and E-cadherin.ResultsThe level of H19 was increased and miR-148b expression was decreased in human AS patient serums and oxidized low-density lipoprotein (ox-LDL)-stimulated human aorta vascular smooth muscle cells (HA-VSMCs). H19 knockdown suppressed proliferation and promoted apoptosis in HA-VSMCs following the treatment of ox-LDL. H19 inhibited miR-148b expression by direct interaction. Moreover, miR-148b inhibitor could reverse the effects of H19 depletion on proliferation and apoptosis in ox-LDL-stimulated HA-VSMCs. Further mechanical explorations showed that WNT1 was a target of miR-148b and H19 acted as a competing endogenous RNA (ceRNA) of miR-148b to enhance WNT1 expression. Furthermore, miR-148 inhibitor exerted its pro-proliferation and anti-apoptosis effects through activating WNT/β-catenin signaling in ox-LDL-stimulated HA-VSMCs.ConclusionH19 facilitated proliferation and inhibited apoptosis through modulating WNT/β-catenin signaling pathway via miR-148b in ox-LDL-stimulated HA-VSMCs, implicating the potential values of H19 in AS therapy.

Inhibition of microRNA‐148b‐3p alleviates oxygen‐glucose deprivation/reoxygenation‐induced apoptosis and oxidative stress in HT22 hippocampal neuron via reinforcing Sestrin2/Nrf2 signalling

“…miR‐148b‐3p functions as a tumour suppressor that inhibits the growth of tumour cells and enhances the sensitivity of tumour cells towards chemo‐ and radiation‐therapy‐induced apoptosis . miR‐148b‐3p inhibition attenuates oxidized low‐density‐lipoprotein‐induced apoptosis in vascular smooth muscle cells . In hepatic sinusoidal endothelial cells, miR‐148b‐3p expression is dysregulated in response to hypoxia treatment and contributes to the modulation of hypoxic injury .…”

LncRNA TNK2‐AS1 regulated ox‐LDL‐stimulated HASMC proliferation and migration via modulating VEGFA and FGF1 expression by sponging miR‐150‐5p

“…The lncRNA myocardial infarction associated transcript (MIAT) was up‐regulated in the serum of the atherosclerotic patients and the ox‐LDL‐treated HASMCs, and lncRNA MIAT overexpression promoted HASMCs proliferation and hindered cell apoptosis via modulating miR‐181b/STAT3 axis . Additionally, lncRNA H19 was increased in the serum of atherosclerotic patients and the ox‐LDL‐stimulated HASMCs, and H19 knockdown inhibited cell proliferation and induced cell apoptosis in ox‐LDL‐stimulated HASMCs via miR‐148/β‐catenin axis . Recently, the lncRNA maternally expressed 8 (MEG8) was identified to be down‐regulated in ox‐LDL‐stimulated VSMCs and enhanced MEG8 expression suppressed VSMC proliferation and migration via regulating miR‐181a/PPARα signalling pathways .…”

NEAT1 contributes to ox‐LDL‐induced inflammation and oxidative stress in macrophages through inhibiting miR‐128

“…LncRNAs have been reported to participate in the pathophysiology of AS . H19 knockdown is able to repress proliferation and induce apoptosis of vascular smooth muscle cells by modulating miR‐148b/WNT/β‐catenin . LncRNA BANCR can facilitate vascular smooth muscle cell proliferation and migration through JNK signaling .…”

“…LncRNA BANCR can facilitate vascular smooth muscle cell proliferation and migration through JNK signaling . LncRNA‐RP11‐714G18.1 depresses vascular cell migration through targeting LRP2BP …”

“…12 H19 knockdown is able to repress proliferation and induce apoptosis of vascular smooth muscle cells by modulating miR-148b/WNT/β-catenin. 13,14 LncRNA BANCR can facilitate vascular smooth muscle cell proliferation and migration through JNK signaling. 15 LncRNA-RP11-714G18.1 depresses vascular cell migration through targeting LRP2BP.…”