slit3 inhibits robo3-induced invasion of synovial fibroblasts in rheumatoid arthritisslit3抑制robo3-induced入侵在类风湿性关节炎滑膜成纤维细胞.pdf

Denk et al. Arthritis Research Therapy 2010, 12:R45 /content/12/2/R45 RESEARCH ARTICLE Open Access Slit3 inhibits Robo3-induced invasion of synovial fibroblasts in rheumatoid arthritis 1 1 2 3 3 1 Alexandra E Denk , Simone Kaufmann , Klaus Stark , Jörg Schedel , Torsten Lowin , Thomas Schubert , Anja K Bosserhoff1* Abstract Introduction: The repellent factor family of Slit molecules has been described to have repulsive function in the developing nervous system on growing axons expressing the Robo receptors. However, until today no data are available on whether these repellent factors are involved in the regulation of synovial fibroblast (SF) activity in rheumatoid arthritis (RA). Methods: mRNA expression in primary synovial fibroblasts was quantified by quantitative reverse transcription PCR and protein expression was measured by fluorescence activated cell sorting (FACS) analysis. Different functional assays were performed with rheumatoid arthritis synovial fibroblasts (RASF): proliferation, migration and a novel in- vitro cartilage destruction assay. Results: First, we found increased expression of Robo3 expression in RASF compared to normal SF. Interestingly, analysis of data from a recently published genome-wide association study suggests a contribution of ROBO3 gene polymorphisms to susceptibility of RA. Functional assays performed with RASF revealed induction of migration and cartilage destruction by Robo3 and increased matrix metalloproteinase (MMP)1 and MMP3 expression. Treatment of RASF in early passages with Slit3 led to inhibition of migration whereas RASF in later passages, having reduced Robo3 expression in cell culture, were not inhibited by Slit3 treatment. Here, reduction of Rob