MicroRNA‑28‑3p inhibits angiotensin‑converting enzyme 2 ectodomain shedding in 293T cells treated with the spike protein of severe acute respiratory syndrome coronavirus 2 by targeting A disintegrin and metalloproteinase 17

Xu,Yun; Li,Yi

doi:10.3892/ijmm.2021.5022

MicroRNA‑28‑3p inhibits angiotensin‑converting enzyme 2 ectodomain shedding in 293T cells treated with the spike protein of severe acute respiratory syndrome coronavirus 2 by targeting A disintegrin and metalloproteinase 17

Authors:
- Yun Xu
- Yi Li
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Affiliations: Department of Infectious Diseases, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China
Published online on: August 16, 2021 https://doi.org/10.3892/ijmm.2021.5022
Article Number: 189
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) is the virus that causes coronavirus disease 2019. Angiotensin‑converting enzyme 2 (ACE2) is the SARS‑CoV binding site and is ubiquitously expressed in endothelial cells of several organs, with the highest levels in the cardiovascular system, kidney and lungs. A disintegrin and metalloproteinase 17 (ADAM17) is involved in ectodomain shedding of ACE2. In the present study, reverse‑transcription‑quantitative PCR, transfection, TUNNEL assay, dual‑luciferase activity assay and western blotting were conducted to investigate the effects of microRNA (miR)‑28‑3p on ADAM17‑dependent shedding of the ACE2 ectodomain following treatment with the spike protein (S‑protein) of SARS‑CoV‑2. It was found that miR‑28‑3p was significantly downregulated in 293T cells treated with 100 ng/ml of S‑protein for 24 h at 37˚C, which led to upregulation of ADAM17. In addition, the expression of ADAM17 and miR‑28‑3p were negatively correlated based on Pearson's correlation test in 293T cells treated with S‑protein for 24 h. Overexpression of miR‑28‑3p and inhibition of ADAM17 regulated 293T cell viability, apoptosis and ACE2 ectodomain shedding. It was also demonstrated that ADAM17 was the target gene of miR‑28‑3p and that miR‑28‑3p negatively regulated ADAM17 expression. Notably, the inhibition of ADAM17 expression blocked the effects of miR‑28‑3p inhibitor on proliferation, apoptosis and ACE2 ectodomain shedding in 293T cells treated with S‑protein. The findings of the present study suggested that miR‑28‑3p inhibits ADAM17‑dependent ACE2 ectodomain shedding in 293T cells treated with the S‑protein of SARS‑CoV‑2, which suggested the potential therapeutic role of miR‑28‑3p mimic in the prevention and treatment of patients with SARS‑CoV‑2.

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