siRNA干扰Rac1表达对人晶状体上皮细胞增殖和迁移能力的影响(1)
[摘要] 目的 研究Rac1对人晶状体上皮细胞SRA01/04增殖和迁移能力的影响，探究Rac1在后囊下混浊及囊袋皱缩综合征发生、发展过程中的作用。 方法 将SRA01/04细胞分为Mock组和siRac1组，构建Rac1相关siRNA并瞬时转染于siRac1组细胞中，Western blot检测Rac1蛋白表达的变化，CCK检测细胞生长周期的变化，流式细胞术检测细胞周期、细胞凋亡变化，Transwell检测Rac1对细胞迁移能力的影响。 结果 两组72 h时吸光度值均高于24 h(P < 0.05)，siRac1组48、72 h时吸光度值均低于Mock组(P < 0.05)；与Mock组比较，siRac1组mRNA和蛋白相对表达量明显降低，G1期细胞所占百分率明显增高，S期细胞所占百分率明显降低，穿越基质膜的细胞数明显减少，差异均有统计学意义(P < 0.05)。F-actin染色结果显示，siRac1组细胞的伪足形成能力较Mock组减弱。 结论 Rac1可能通过调节晶状体上皮细胞的增殖能力和迁移能力，影响后囊下混浊及囊袋皱缩综合征的发生、发展。
[关键词] 后囊下混浊；囊袋皱缩综合征；Rac1；晶状体；上皮细胞
[中图分类号] R776.1 [文献标识码] A [文章编号] 1673-7210(2018)10(b)-0017-04
[Abstract] Objective To study the effect of Rac1 on the proliferation and migration of human lens epithelial cells SRA01/04， and to explore the role of Rac1 in the development and progression of posterior capsular opacities and capsular contraction syndrome. Methods SRA01/04 cells were divided into mock group and siRac1 group. Rac1-related siRNA was constructed and transiently transfected in siRac1 group cells. Western blot was used to detect the changes of Rac1 protein expression， CCK was used to detect the changes of cell growth cycle， flow cytometry was used to detect the cell cycle and apoptosis， Transwell was used to detect the effect of Rac1 on cell migration ability. Results The absorbance values of the two groups at 72 hours were higher than those at 24 hours (P < 0.05)， the absorbance values of siRac1 group at 48， 72 hours were lower than those of mock group (P < 0.05). Compared with mock group， the relative expression of mRNA and protein in siRac1 group decreased significantly， the percentage of G1 phase cells increased significantly， while the percentage of S phase cells decreased significantly， the number of cells passing through the membrane reduced significantly， the differences were statistically significant (P < 0.05). F-actin staining showed that the forming ability of pseudopodium in siRac1 group was significantly weaker than that of mock group. Conclusion Rac1 may affect the occurrence and development of posterior capsular opacities and capsular contraction syndrome through adjusting the proliferation and migration of lens epithelial cells.
[Key words] Posterior capsular opacities； Capsular contraction syndrome； Rac1； Lens； Epithelial cells
后囊下混濁(posterior capsular opacities，PCO)和囊袋皱缩综合征(capsular contraction syndrome，CCS)是白内障超声乳化术后常见并发症，其发生与发展均与白内障术后残留晶状体上皮细胞的增殖与迁移有关[1-2]。Rac1是Rho家族重要成员之一，有促进细胞迁移、抑制细胞凋亡的作用[3]。近年来有文献证实，Rac1可以促进大鼠视网膜新生血管的形成[4]，而且在人翼状胬肉也有表达[5]。但Rac1作用于晶状体上皮细胞的实验研究较少。我们在人晶状体上皮细胞SRA01/04中通过siRNA来干扰Rac1的表达以观察其对细胞增殖和迁移能力的影响，从而验证通过降低Rac1表达以降低PCO和CCS发生及进展的可能性。, 百拇医药(孙国庆 唐建明)
[关键词] 后囊下混浊；囊袋皱缩综合征；Rac1；晶状体；上皮细胞
[中图分类号] R776.1 [文献标识码] A [文章编号] 1673-7210(2018)10(b)-0017-04
[Abstract] Objective To study the effect of Rac1 on the proliferation and migration of human lens epithelial cells SRA01/04， and to explore the role of Rac1 in the development and progression of posterior capsular opacities and capsular contraction syndrome. Methods SRA01/04 cells were divided into mock group and siRac1 group. Rac1-related siRNA was constructed and transiently transfected in siRac1 group cells. Western blot was used to detect the changes of Rac1 protein expression， CCK was used to detect the changes of cell growth cycle， flow cytometry was used to detect the cell cycle and apoptosis， Transwell was used to detect the effect of Rac1 on cell migration ability. Results The absorbance values of the two groups at 72 hours were higher than those at 24 hours (P < 0.05)， the absorbance values of siRac1 group at 48， 72 hours were lower than those of mock group (P < 0.05). Compared with mock group， the relative expression of mRNA and protein in siRac1 group decreased significantly， the percentage of G1 phase cells increased significantly， while the percentage of S phase cells decreased significantly， the number of cells passing through the membrane reduced significantly， the differences were statistically significant (P < 0.05). F-actin staining showed that the forming ability of pseudopodium in siRac1 group was significantly weaker than that of mock group. Conclusion Rac1 may affect the occurrence and development of posterior capsular opacities and capsular contraction syndrome through adjusting the proliferation and migration of lens epithelial cells.
[Key words] Posterior capsular opacities； Capsular contraction syndrome； Rac1； Lens； Epithelial cells
后囊下混濁(posterior capsular opacities，PCO)和囊袋皱缩综合征(capsular contraction syndrome，CCS)是白内障超声乳化术后常见并发症，其发生与发展均与白内障术后残留晶状体上皮细胞的增殖与迁移有关[1-2]。Rac1是Rho家族重要成员之一，有促进细胞迁移、抑制细胞凋亡的作用[3]。近年来有文献证实，Rac1可以促进大鼠视网膜新生血管的形成[4]，而且在人翼状胬肉也有表达[5]。但Rac1作用于晶状体上皮细胞的实验研究较少。我们在人晶状体上皮细胞SRA01/04中通过siRNA来干扰Rac1的表达以观察其对细胞增殖和迁移能力的影响，从而验证通过降低Rac1表达以降低PCO和CCS发生及进展的可能性。, 百拇医药(孙国庆 唐建明)