Knockdown of NCK1-AS1 inhibits the development of atherosclerosis by targeting miR-1197/COX10 axis

• Published in: Journal of biological engineering Vol. 16; no. 1; p. 2
• Main Authors: Zhang, Bin, Wang, Juncheng, Du, Lei, Shao, Lufei, Zou, Yourui, Liu, Haibo, Liu, Jinfang
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 05.01.2022; BioMed Central; BMC
• Subjects: Analysis; Antibodies; Apoptosis; Arteriosclerosis; Assaying; Atherosclerosis; Binding sites; Blood; Cancer; Cell growth; Cell viability; Cholecystokinin; COX10; Cytokines; Enzyme-linked immunosorbent assay; Evaluation; Experiments; Health aspects; IL-1β; Inflammation; Interleukin 6; Low density lipoprotein; Low density lipoproteins; Malondialdehyde; miR-1197; Muscles; NCK1-AS1; Non-coding RNA; Oxidative stress; Phenotypes; Polymerase chain reaction; Proteins; RNA; Smooth muscle; Statistical analysis; Therapeutic targets; Transfection; Variance analysis; China; Beijing China; United Kingdom > UK; United States > US; Germany; miR-1197; NCK1-AS1; COX10; Atherosclerosis
• ISSN: 1754-1611; 1754-1611
• DOI: 10.1186/s13036-021-00281-6

Although long non-coding RNA (lncRNA) NCK1-AS1 plays important roles in human cancer, its function in atherosclerosis (AS) remains unclear. The expression of NCK1-AS1 in AS blood samples was detected by qRT-PCR. Oxidized low-density lipoprotein (ox-LDL) was used to construct the AS cell model, and quantitative real-time polymerase chain reaction (qRT-PCR) assay was used to evaluate NCK1-AS1 level. Cell phenotypes including proliferation and apoptosis were assessed by Cell Counting Kit-8 (CCK-8) assay and flow cytometer, respectively. The malondialdehyde level was measured to evaluate oxidative stress. The expression of apoptosis-related proteins was evaluated by western blot. The expression of inflammatory cytokines (IL-1β, IL-6 and TNK-α) was measured by qRT-PCR and ELISA assays. The relationship among NCK1-AS1, miR-1197 and COX10 was determined by bioinformatic analysis and luciferase reporter assay. NCK1-AS1 was significantly upregulated in AS blood samples and ox-LDL stimulated vascular smooth muscle cells (VSMCs). Knockdown of NCK1-AS1 increased cell viability, reduced cell apoptosis and MDA level, and also inhibited the expression of inflammatory cytokines (IL-1β, IL-6 and TNK-α) in ox-LDL stimulated VSMCs. NCK1-AS1 could positively regulate COX10 expression by directly sponging miR-1197. Moreover, co-transfection of sh-NCK1-AS1 and miR-1197 inhibitor, or co-transfection of sh-NCK1-AS1 and pc-COX10 (COX10 overexpressing plasmid) obviously reduced cell viability, promoted cell apoptosis, and increased MDA level in VSMCs followed by ox-LDL treatment for 24 h compared to that in sh-NCK1-AS1 transfected VSMCs. Our study revealed that knockdown of NCK1-AS1 attenuated the development of AS by regulating miR-1197/COX10 axis, suggesting that this lncRNA might be a potential therapeutic target for AS.