Downregulated miR‑302d‑3p promotes chondrocyte proliferation and migration by regulation of Unc‑51‑like kinase 1

Wang,Shangzeng; Zheng,Yongzhi; Hu,Zheng; Wang,Zhen; Zhang,Yingjie; Wei,Licheng

doi:10.3892/ijmm.2019.4267

Downregulated miR‑302d‑3p promotes chondrocyte proliferation and migration by regulation of Unc‑51‑like kinase 1

Authors:
- Shangzeng Wang
- Yongzhi Zheng
- Zheng Hu
- Zhen Wang
- Yingjie Zhang
- Licheng Wei
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Affiliations: Department of Orthopedics, The Second Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan 450002, P.R. China, Department of Orthopedics, Changsha Hospital of Traditional Chinese Medicine, Changsha Eighth Hospital, Changsha, Hunan 410100, P.R. China, Graduate School of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan 450003, P.R. China
Published online on: July 5, 2019 https://doi.org/10.3892/ijmm.2019.4267
Pages: 1039-1047
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteoarthritis (OA) is a common musculoskeletal disease and is related to the function of chondrocytes. The aim of the present study was to investigate the effects of miR‑302d‑3p on chondrocytes. Quantitative PCR (qPCR) was conducted to detect mRNA expression, while western blotting was performed to investigate protein expression in these cells. RNAs mimics, inhibitors and small interfering (si)RNAs were respectively transfected into chondrocytes (CHON‑001 cell line), after which, a Cell Counting Kit‑8 assay was performed to detect chondrocyte viability. Giemsa staining of the cells was also conducted to analyze the colony formation ability of the cells. Additionally, cell apoptosis was evaluated with an apoptosis detection kit using flow cytometry. A scratch‑wound assay was conducted to investigate cell migration. Bioinformatics analysis using TargetScan 7.2 revealed the potential the target gene of microRNA (miR)‑302d‑3p; a dual luciferase reporter assay determined the target gene. Suppression of miR‑302d‑3p increased the viability of cells, cell colony number and migration; CHON‑001 cell apoptosis was also inhibited. miR‑302d‑3p mimics decreased the luciferase activity of reporter plasmids containing the wild-type 3'‑untranslated region of Unc‑51‑like kinase 1 (ULK1). siULK1 decreased CHON‑001 cell viability and migration. Furthermore, siULK1 promoted the expression of phosphorylated IκΒα and p65, while miR‑302d‑3p inhibitor suppressed the expression of phosphorylated IκΒα and p65. Inhibition of miR‑302d‑3p could promote the proliferation and migration, and inhibit the apoptosis of chondrocytes, potentially by upregulating ULK1; thus, inflammation may be suppressed. The findings of the present study suggest miR‑302d‑3p and ULK1 as potential therapeutic targets for the prevention and treatment of OA.

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