The lncRNA DANCR promotes development of atherosclerosis by regulating the miR-214-5p/COX20 signaling pathway

Ruolan Zhang,Yuming Hao,Jinrong Zhang

doi:10.1186/s11658-022-00310-2

Abstract

BackgroundAlthough long non-coding RNA differentiation antagonizing non-protein coding RNA (DANCR) has been reported to be involved in atherosclerosis (AS) development, its specific mechanism remains unclear.MethodsDANCR expression levels in blood samples of AS patients and oxidized low-density lipoprotein (ox-LDL) treated vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The small interfering RNA targeting DANCR (si-DANCR) was used to silence DANCR expression. Cell viability was assessed by CCK-8 assay. Cell apoptosis was evaluated by flow cytometry. Levels of inflammatory cytokines, anti-oxidative enzyme superoxide dismutase (SOD) activity, and malonaldehyde (MDA) were detected by specific commercial kits. An animal AS model was established to confirm the role of DANCR/microR-214-5p/COX20 (the chaperone of cytochrome c oxidase subunit II COX2) in AS development.ResultsDANCR was significantly increased in the blood samples of AS patients and ox-LDL treated VSMCs and HUVECs. DANCR downregulation obviously increased viability and reduced apoptosis of ox-LDL-treated VSMCs and HUVECs. Meanwhile, DANCR downregulation reduced the levels of inflammatory cytokines, including interleukin (IL)-6 (IL-6), IL-1beta (IL-1β), IL-6 and tumor necrosis factor (TNF)-alpha (TNF-α) and MDA while increasing the SOD level in ox-LDL-treated VSMCs and HUVECs. DANCR regulated COX20 expression by acting as a competing endogenous RNA (ceRNA) of miR-214-5p. Rescue experiments demonstrated that miR-214-5p downregulation obviously attenuated si-DANCR-induced protective effects on ox-LDL-caused endothelial injury.ConclusionsOur results revealed that DANCR promoted AS progression by targeting the miR-214-5p/COX20 axis, suggesting that DANCR might be a potential therapeutic target for AS.

Highlights

Atherosclerosis (AS) is a chronic inflammatory disorder and the most common leading cause of cerebrovascular diseases (CVD) [1, 2]
We confirmed that differentiation antagonizing nonprotein coding RNA (DANCR) was significantly upregulated in the blood samples of AS patients and oxidized low-density lipoprotein (ox-LDL) treated vascular smooth muscle cells (VSMCs) and human umbilical vein endothelial cells (HUVECs)
We stimulated VSMCs and HUVECs with ox-LDL (25, 50, and 100 μg/ml) for 48 h and found that DANCR was significantly upregulated in ox-LDL treated cells (Fig. 1B)

Summary

Introduction

Atherosclerosis (AS) is a chronic inflammatory disorder and the most common leading cause of cerebrovascular diseases (CVD) [1, 2]. It has been reported that endothelial cell dysfunction is an essential process for AS progression [4]. Understanding the specific molecular mechanisms involved in ox-LDL-induced endothelial cell dysfunction may contribute to developing efficient diagnostic and therapeutic targets for AS. Long non-coding RNAs (lncRNAs), a type of RNA transcripts more than 200 nucleotides in length, have been reported to participate in various biological and pathological processes [6]. Differentiation antagonizing non-protein coding RNA (DANCR), a widely studied lncRNA, has been reported to participate in the development of various human cancers such as non-small cell lung cancer [11] and colorectal cancer [12]. Long non-coding RNA differentiation antagonizing nonprotein coding RNA (DANCR) has been reported to be involved in atherosclerosis (AS) development, its specific mechanism remains unclear

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Results

Conclusion