LncRNA RASSF8-AS1 knockdown displayed antiproliferative and proapoptotic effects through miR-188-3p/ATG7 pathway in ox-LDL-treated vascular smooth muscle cells.

Abstract

Long noncoding RNA (lncRNA)-mediated changes in gene expression contribute to atherosclerosis (AS) development. However, the roles of numerous lncRNAs in AS have not been fully elucidated. Here, we aimed to investigate the potential role of lncRNA RASSF8-AS1 (RASSF8-AS1) in autophagy of human aortic vascular smooth muscle cells (HA-VSMCs). RASSF8-AS1 expression in patients with AS was extracted from the Gene Expression Omnibus (GEO) database. RASSF8-AS1 and microRNA-188-3p (miR-188-3p) expression was analyzed in 20 enrolled patients with AS. HA-VSMCs were treated with oxidized low-density lipoprotein (ox-LDL) (25, 50, 75, and 100 µg/mL) for 24 h. Loss- or gain-of-function of RASSF8-AS1, miR-1883p, and autophagy-related 7 (ATG7) was studied using the transfected HA-VSMCs. Cell viability was assessed using Cell Counting Kit-8 (CCK-8). Apoptosis was detected with annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI). Relative luciferase reporter assay was used to confirm the targeting relationship of miR-188-3p to RASSF8-AS1 or ATG7. Gene expression was detected by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot. RASSF8-AS1 was enriched in the serum of patients with AS and ox-LDL-treated HA-VSMCs. Ox-LDL induced proliferation and autophagy while inhibiting the apoptosis of HA-VSMCs, which was abated by RASSF8-AS1 knockdown. RASSF8-AS1 downregulated miR-188-3p of ox-LDL-treated HA-VSMCs. RASSF8-AS1 knockdown caused an increase in miR-188-3p, which inhibited proliferation and autophagy and induced the apoptosis of ox-LDL-treated HA-VSMCs. miR-188-3p inhibited ATG7 expression in ox-LDL-treated HA-VSMCs. RASSF8-AS1 elevated ATG7 and induced autophagy through sponging miR-188-3p in ox-LDL-treated HA-VSMCs. RASSF8-AS1 regulated autophagy by targeting miR-188-3p, a messenger RNA-binding miRNA that increases ATG7 level, which may be a new target molecule for the prevention and prognosis of AS.