title: 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.
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.
authors.affiliation: Department of Rheumatology, The First Affiliated Hospital of China Medical University, 2 Zhongshan Road, Shenyang, 110001, People's Republic of China.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Yuan
authors.forename: Pingting
authors.initials: P
authors.lastname: Yang
authors.forename: Dun
authors.initials: D
authors.lastname: Zhou
authors.forename: Wei
authors.initials: W
authors.lastname: Gao
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Qiu
authors.forename: Fang
authors.initials: F
authors.lastname: Fang
authors.forename: Shuang
authors.initials: S
authors.lastname: Ding
authors.forename: Weiguo
authors.initials: W
authors.lastname: Xiao
mesh_terms: D001172:Arthritis, Rheumatoid / Q000235:genetics*; D002465:Cell Movement; D002478:Cells, Cultured; D002867:Chromones / Q000494:pharmacology; D015536:Down-Regulation; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D009025:Morpholines / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D013583:Synovial Membrane / Q000166:cytology* / Q000378:metabolism; D000099157:Sphingosine Kinase
chemical_list: D002867:Chromones; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D009025:Morpholines; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D051057:Proto-Oncogene Proteins c-akt; D034741:RNA, Small Interfering; D000099157:Sphingosine Kinase; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C522361:MMP2 protein, human; C579270:MMP9 protein, human