Roles of layilin in human synovial fibroblasts revealed by proteomic analysis
Roles of layilin in human synovial fibroblasts revealed by proteomic analysis
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DOI：
10.15761/imm.1000316
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
K. Shimazaki;M. Arito;Toshiyuki Sato;K. Omoteyama;Masaaki Sato;M. Kurokawa;N. Suematsu;H. Niki;Tomohiro Kato
中科院分区：
文献类型：
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作者：
K. Shimazaki;M. Arito;Toshiyuki Sato;K. Omoteyama;Masaaki Sato;M. Kurokawa;N. Suematsu;H. Niki;Tomohiro Kato
目的：为了了解layilin在滑膜成纤维细胞中的生理和病理作用，我们全面研究layilin沉默引起的蛋白谱变化。方法：在TNF-α-处理和TNF-α-未处理的条件下，转染永生化人滑膜成纤维细胞（HSFs）的siRNA。采用二维荧光差凝胶电泳（2D-DIGE）全面检测受siL影响的蛋白。用质谱法鉴定感兴趣的蛋白质。结果：在2D-DIGE分析中，在TNF-α-处理和-α-未处理的情况下，大约五分之一的检测蛋白斑点的强度被siL显著改变（分别为239/1092点和201/1092点）。在87个蛋白点中，有25个蛋白被siL鉴定为±1.3倍或更大的强度变化。24个蛋白点中有15个（62.5%）为上皮-间质转化（EMT）相关蛋白。结论：我们的数据表明layilin深入参与emt相关蛋白的调控。虽然滑膜成纤维细胞不是上皮细胞，但在EMT的背景下，应该研究layilin在滑膜成纤维细胞中的功能。此外，受layilin影响的emt相关蛋白可能参与了以滑膜成纤维细胞增生和侵袭为特征的类风湿关节炎的发病机制。
Aim: To understand physiological and pathological roles of layilin in synovial fibroblasts, we comprehensively investigated protein profile changes caused by layilinsilencing. Methods: Immortalized human synovial membrane fibroblasts (HSFs) were transfected with siRNA for layilin (siL) in TNF-α-treated and -non-treated conditions. Proteins affected by siL were comprehensively detected by 2-dimensional fluorescence difference gel electrophoresis (2D-DIGE). Proteins of interest were identified by mass spectrometry. Results: In the 2D-DIGE analysis, intensity of approximately one-fifth of the detected protein spots was significantly changed by siL both in the TNF-α-treated and –non-treated conditions (239/1092 spots and 201/1092 spots, respectively). Proteins were identified in 25 out of 87 protein spots with ± 1.3-fold or greater intensity changes by siL. 15 (62.5%) of the 24 protein spots were assigned to epithelial-mesenchymal transition (EMT)-related proteins. Conclusion: Our data suggest that layilin is deeply involved in the regulation of EMT-related proteins. Functions of layilin in synovial fibroblasts should be investigated in the context of EMT, although synovial fibroblasts are not epithelial cells. Furthermore, the EMT-related proteins affected by layilin may be involved in the pathogenesis of rheumatoid arthritis, which was characterized by proliferation and invasion of synovial fibroblasts.