a prokaryotic s1p lyase degrades extracellular s1p in vitro and in vivo implication for treating hyperproliferative disorders一个原核s1p裂合酶降解细胞外s1p体外和体内暗示治疗hyperproliferative紊乱.pdfVIP

A Prokaryotic S1P Lyase Degrades Extracellular S1P In Vitro and In Vivo: Implication for Treating Hyperproliferative Disorders 1 2 1 1 3 Andrea Huwiler , Florence Bourquin , Nataliya Kotelevets , Oleksandr Pastukhov , Guido Capitani , ¨ 2, Uwe Zangemeister-Wittke1,2* Markus G. Grutter ¨ ¨ 1 Institute of Pharmacology, University of Bern, Bern, Switzerland, 2 Institute of Biochemistry, University of Zurich, Zurich, Switzerland, 3 Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland Abstract Sphingosine-1-phosphate (S1P) regulates a broad spectrum of fundamental cellular processes like proliferation, death, migration and cytokine production. Therefore, elevated levels of S1P may be causal to various pathologic conditions including cancer, fibrosis, inflammation, autoimmune diseases and aberrant angiogenesis. Here we report that S1P lyase from the prokaryote Symbiobacterium thermophilum (StSPL) degrades extracellular S1P in vitro and in blood. Moreover, we investigated its effect on cellular responses typical of fibrosis, cancer and aberrant angiogenesis using renal mesangial cells, endothelial cells, breast (MCF-7) and colon (HCT 116) carcinoma cells as disease models. In all cell types, wild-type StSPL, but not an inactive mutant, disrupted MAPK phosphorylation stimulated by exogenous S1P. Functionally, disruption of S1P receptor signaling by S1P depletion inhibited proliferation and expression of connective tissue growth factor in mesangial cells, proliferation, migration and VEGF expression in carcinoma cells, and proliferation and m