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Knockdown of sphingosine kinase 1 inhibits the migration and invasion of human rheumatoid arthritis fibroblast-like synoviocytes by down-regulating the PI3K/AKT activation and MMP-2/9 production in vitro

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Abstract

To investigate the potential regulation of sphingosine kinase 1 (SPHK1) on the migration, invasion, and matrix metalloproteinase (MMP) expression in human rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). RA-FLS were transfected control siRNA or SPHK1 siRNA. The migration and invasion of unmanipulated control, control siRNA or SPHK1 siRNA- transfected RA-FLS in vitro were measured by the transwell system. The relative levels of SPHK1, PI3K, and AKT as well as AKT phosphorylation in RA-FLS were determined by Western blot. The levels of MMP-2/9 secreted by RA-FLS were detected by ELISA. Knockdown of SPHK1 significantly inhibited the spontaneous migration and invasion of RA-FLS, accompanied by significantly reduced levels of PI3K expression and AKT phosphorylation. Similarly, treatment with LY294002, an inhibitor of the PI3K/AKT pathway, inhibited the migration and invasion of RA-FLS. Knockdown of SPHK1 and treatment with the inhibitor synergistically inhibited the migration and invasion of RA-FLS, by further reducing the levels of PI3K expression and AKT phosphorylation. In addition, knockdown of SPHK1 or treatment with LY294002 inhibited the secretion of MMP-2 and MMP-9, and both synergistically reduced the production of MMP-2 and MMP-9 in RA-FLS in vitro. Knockdown of SPHK1 expression inhibits the PI3K/AKT activation, MMP-2 and MMP-9 expression, and human RA-FLS migration and invasion in vitro. Potentially, SPHK1 may be a novel therapeutic target for RA.

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Acknowledgments

This work was sponsored by a grant from the Education Project of Liaoning Province (No. L2012312). The Authors thank Medjaden Bioscience Limited for editing and proofreading this manuscript.

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We declare that we have no conflicts of interest.

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Authors and Affiliations

  1. Department of Rheumatology, The First Affiliated Hospital of China Medical University, 2 Zhongshan Road, Shenyang, 110001, People’s Republic of China

    Hongxia Yuan, Pingting Yang, Fang Fang, Shuang Ding & Weiguo Xiao

  2. Liaoning Medical University, 40 Songpo Road, Jinzhou, 121001, People’s Republic of China

    Hongxia Yuan, Dun Zhou, Wei Gao & Zhenyu Qiu

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  1. Hongxia Yuan
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Yuan, H., Yang, P., Zhou, D. et al. Knockdown of sphingosine kinase 1 inhibits the migration and invasion of human rheumatoid arthritis fibroblast-like synoviocytes by down-regulating the PI3K/AKT activation and MMP-2/9 production in vitro. Mol Biol Rep 41, 5157–5165 (2014). https://doi.org/10.1007/s11033-014-3382-4

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