Abstract
Osteoarthritis is a heterogeneous disease with a complex etiology. However, there is no effective treatment strategy at present. The purpose of this study was to explore the miRNA‒mRNA regulatory network and molecular mechanism that regulate the progression of osteoarthritis. In this article, we downloaded datasets (GSE55457, GSE82107, GSE143514 and GSE55235) from Gene Expression Omnibus (GEO) to screen differentially expressed mRNAs in osteoarthritis. Then, through weighted gene coexpression network (WGCNA), functional enrichment, protein‒protein interaction (PPI) network, miRNA‒mRNA coexpression network, ROC curve, and immune infiltration analyses and qPCR, the mRNA PLCD3, which was highly expressed in osteoarthritis and had clinical predictive value, was screened. We found that PLCD3 directly targets miR-34a-5p through DIANA and dual-luciferase experiments. The expression levels of PLCD3 and miR-34a-5p were negatively correlated. In addition, CCK-8 and wound healing assays showed that the miR-34a-5p mimic inhibited hFLS-OA cell proliferation and promoted hFLS-OA cell migration. PLCD3 overexpression showed the opposite trend. Western blotting further found that overexpression of miR-34a-5p reduced the protein expression levels of p-PI3K and p-AKT, while overexpression of PLCD3 showed the opposite trend. In addition, combined with the effect of the PI3K/AKT pathway inhibitor BIO (IC50 = 5.95 μM), the results showed that overexpression of miR-34a-5p increased the inhibitory effects of BIO on p-PI3K and p-AKT protein expression, while overexpression of PLCD3 significantly reversed these inhibitory effects. Overall, the miR-34a-5p/PLCD3 axis may mediate the PI3K/AKT pathway in regulating cartilage homeostasis in synovial osteoarthritis. These data indicate that miR-34a-5p/PLCD3 may be a new prognostic factor in the pathology of synovial osteoarthritis.
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Data availability
Expression profile data (GSE55457, GSE82107, GSE143514 and GSE55235) in this study were from the public database GEO (https://www.ncbi.nlm.nih.gov/geo). All datasets generated for this study are included in the article and supplementary material.
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Acknowledgements
We thank all GEO data builders and data contributors, as well as the team that built the DIANA Tools online analysis page. Thanks are also due to Shanghai Ordovician Biotechnology Co., Ltd. for providing the platform for the biological information analysis.
Funding
This study was funded by the Youth Project “Science and Education” of Suzhou (KJXW2020068), the Basic Research Project of Changzhou Science and Technology Bureau (CJ20200112), and the Project of Changshu Hospital Affiliated to Nanjing University of Chinese Medicine (cszyy201910).
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YP conducted the data analysis and wrote the manuscript. XY, JXW, XYi, WQ, DWG and WKJ carried out the data analysis and manuscript revision. DXY designed the study. All authors read and approved the final manuscript.
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Ying, P., Xu, Y., Jiang, X. et al. Analysis of the regulatory role of miR-34a-5p/PLCD3 in the progression of osteoarthritis. Funct Integr Genomics 23, 131 (2023). https://doi.org/10.1007/s10142-023-01058-4
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DOI: https://doi.org/10.1007/s10142-023-01058-4