hsa‑miR‑15b‑5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis

Gan,Shujie; Mao,Jieqi; Pan,Yuqin; Tang,Jingdong; Qiu,Zhengjun

doi:10.3892/etm.2021.10642

hsa‑miR‑15b‑5p regulates the proliferation and apoptosis of human vascular smooth muscle cells by targeting the ACSS2/PTGS2 axis

Authors:
- Shujie Gan
- Jieqi Mao
- Yuqin Pan
- Jingdong Tang
- Zhengjun Qiu
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Affiliations: Department of Vascular Surgery, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China, Department of Nursing, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, P.R. China, Department of Vascular Surgery, Shanghai Pudong Hospital Affiliated to Fudan University, Shanghai 201399, P.R. China
Published online on: August 24, 2021 https://doi.org/10.3892/etm.2021.10642
Article Number: 1208
Copyright: © Gan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A previous bioinformatic analysis from our group predicted that the interaction of microRNA (miRNA/miR)‑15b with the acyl‑CoA synthetase short chain family member 2 (ACSS2) gene was important for the development of abdominal aortic aneurysm (AAA). Apoptosis of aortic vascular smooth muscle cells (VSMCs) is a pathological feature of AAA. The present study aimed to explain the roles of miR‑15b/ACSS2 in AAA by exploring their effects on the proliferation and apoptosis of aortic VSMCs. Human aortic VSMCs (T/G HA‑VSMC cell line) were divided into six groups and were transfected with miR‑15b‑5p mimics, mimic negative control (NC), miR‑15b‑5p inhibitors, inhibitor NC, miR‑15b‑5p mimics+pcDNA3.1 and miR‑15b‑5p mimics+ACSS2‑overexpessing vector. CCK‑8 assay was used to determine cell proliferation. Annexin V‑FITC/PI staining and flow cytometry assays were used to measure cell apoptosis. Dual‑luciferase reporter assays were used to confirm the targeted relationship between miR‑15b‑5p and ACSS2. Reverse transcription‑quantitative PCR and/or western blotting were used to examine the expression levels of miR‑15b‑5p, ACSS2 and prostaglandin‑endoperoxide synthase 2 (PTGS2). Following transfection of T/G HA‑VSMCs with mimics and inhibitors to respectively upregulate and downregulate miR‑15b‑5p, the results demonstrated that overexpression of miR‑15b‑5p inhibited cell proliferation and promoted cell apoptosis; silencing of miR‑15b‑5p obtained the opposite results. ACSS2 may be a direct target of miR‑15b‑5p, since the luciferase activity of a ACSS2 wild‑type vector, but not that of a ACSS2 mutant reporter, was significantly inhibited by miR‑15b‑5p mimics compared with controls. Additionally, the expression levels of ACSS2 and its downstream gene PTGS2 were significantly reduced or increased following transfection with miR‑15b‑5p mimics or inhibitors, respectively. Furthermore, overexpression of ACSS2 reversed the antiproliferative and proapoptotic effects of miR‑15b‑5p mimics by blocking the production of PTGS2 protein. In conclusion, miR‑15b‑5p may promote the apoptosis and inhibit the proliferation of aortic VSMCs via targeting the ACSS2/PTGS2 axis. The present study provided preliminary evidence indicating that the miR‑15b‑5p/ACSS2/PTGS2 axis may be a potential target for the treatment of AAA.

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