下调Smad1基因抑制RA-FLS侵袭和迁移

Down-regulation of Smad1 Gene Inhibits RA-FLS Invasion and Migration

目的 探讨Smad1基因对类风湿关节炎滑膜成纤维细胞侵袭和迁移的影响及机制.方法 将Smad1 siRNA转染MH7A细胞,实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白免疫印迹(Western blot)法检测干涉效率.Western blot法检测各组细胞中上皮细胞钙黏蛋白(E-cadherin)、波形蛋白(vimentin)和α平滑肌肌动蛋白(α-SMA)及基质金属蛋白酶-3(MMP-3)和MMP-13表达水平,Transwell实验分析MH7A细胞侵袭和迁移能力.结果 转染Smad1 siRNA可有效抑制MH7A细胞中Smad1 mRNA和蛋白的表达.抑制Smad1的表达后,α-SMA和vimentin表达水平显著下调,E-cadherin的表达明显上调;MH7A细胞上皮间质转化(EMT)过程受到阻碍,侵袭和迁移能力均受到抑制,MH7A细胞中MMP-3和MMP-13表达降低.结论 下调Smad1基因表达,可降低MMP-3和MMP-13的表达水平,进而阻碍EMT的发生,抑制MH7A细胞的体外侵袭和迁移.

Objective To investigate the effect and mechanism of Smad1 gene on the invasion and migration of synovial fibroblasts in rheumatoid arthritis. Methods Smad1 siRNA was transfected into MH7 A cells. The interference efficiency was detected by qRT-PCR and Western blot. Western blot was used to detect E-cadherin, vimentin and α-SMA, MMP-3 and MMP-13 expression levels. Transwell assay to detect cell invasion and migration ability. Results Smad1 siRNA can effectively inhibit the expression of Smad1 mRNA and protein in MH7 A cells. After inhibiting the expression of Smad1, the expression levels of α-SMA and vimentin were significantly down-regulated, the expression of E-cadherin was significantly up-regulated, the process of epithelial-mesenchymal transition(EMT) in MH7 A cells is hindered.Invasion and migration capabilities were suppressed. MMP-3 and MMP-13 expression was decreased in MH7 A cells.Conclusion Down-regulation of Smad1 gene expression can reduce the expression levels of MMP-3 and MMP-13, thereby impedes the occurrence of EMT and inhibites the invasion and migration of MH7 A cells in vitro.