Institute of Pathology, University of Regensburg, Regensburg, Germany.
Int J Mol Med. 2012 Nov;30(5):1133-7. doi: 10.3892/ijmm.2012.1098. Epub 2012 Aug 20.
The repellent factor family of Slit molecules has been described as having a repulsive function in the developing nervous system on growing axons expressing the Roundabout (Robo) receptors. Recent studies determined the effects of Slit molecules on the migratory and invasive potential of several types of tumor cells but also on synovial fibroblasts (SFs) derived from rheumatoid arthritis (RA) patients. To optimize a potential therapeutic application we aimed at generatingfragments of Slit3 showing the same functional ability as the full-length molecule but having the advantage of a smaller size. Recombinant Slit3 proteins were expressed and analyzed by western blotting. Their activity was defined by functional assays such as migration assays with RASF and melanoma cells. Recombinant Slit3 containing only leucine rich repeat domain 2 (D2), the domain important for Robo binding and the minimal functional unit D2 dNC were both able to inhibit migration of RASFs as effectively as Slit3 with all 4 repeats. Collectively, our data showed that the ability of Slit3 to reduce the migratory activity of synovial cells from patients with RA and melanoma cells can be mimicked by small protein fragments derived from Slit3. Slit3 fragments may be helpful in therapeutic attempts; however, further studies are necessary in order to elucidate their activity in vivo.
Slit 分子的排斥因子家族被描述为在表达 Roundabout（Robo）受体的生长轴突的发育神经系统中具有排斥功能。最近的研究确定了 Slit 分子对几种类型的肿瘤细胞的迁移和侵袭潜力的影响，但也对源自类风湿关节炎（RA）患者的滑膜成纤维细胞（SFs）有影响。为了优化潜在的治疗应用，我们旨在生成具有与全长分子相同功能能力但具有更小尺寸优势的 Slit3 片段。通过 Western blot 分析表达和分析重组 Slit3 蛋白。通过迁移测定等功能测定来定义其活性，例如与 RASF 和黑素瘤细胞的迁移测定。仅含有亮氨酸重复结构域 2（D2）的重组 Slit3，该结构域对于 Robo 结合很重要，并且是最小的功能单元 D2 dNC，均能够像具有所有 4 个重复的 Slit3 一样有效地抑制 RASF 的迁移。总之，我们的数据表明，Slit3 减少 RA 患者滑膜细胞和黑素瘤细胞迁移活性的能力可以被源自 Slit3 的小蛋白片段模拟。Slit3 片段可能有助于治疗尝试；然而，为了阐明它们在体内的活性，还需要进一步的研究。
