c-Fos基因RNA干扰对大鼠血管平滑肌细胞表型及迁移的影响  

Effect of RNA Interference for c-Fos Gene on the Phenotypic Transition and Migration of Vascular Smooth Muscle Cells

作　　者：张宏伟[1] 刘宇[2] 杨野[2] 张强[2] 

机构地区：[1]天津东丽医院普外科,天津300300 [2]中国医科大学附属一院干诊科,沈阳110001

出　　处：《中国医科大学学报》2011年第8期691-695,共5页Journal of China Medical University

基　　金：黑龙江省卫生厅科研课题(2009-320)

摘　　要：目的以RNA干扰抑制血管平滑肌细胞(VSMC)c-Fos基因,探讨c-Fos基因表达抑制后c-Fos siRNA对平滑肌细胞表型转换及迁移的作用。方法设置正常对照组、阴性siRNA组及阳性c-Fos siRNA转染组。应用RT-PCR半定量法检测VSMC中c-Fos和α平滑肌肌动蛋白(α-SM-actin)mRNA的水平,采用Western blot检测α-SM-actin、碱性成纤维细胞生长因子(bFGF)、内皮素1(ET-1)蛋白活性的变化,应用免疫荧光染色检测平滑肌分化标志蛋白α-SM-actin,同时应用Transwell检测平滑肌细胞迁移能力。结果 siRNA转染后阳性c-Fos siRNA转染组c-Fos基因表达水平降低,而正常对照组与阴性siRNA组比较c-Fos基因表达水平无统计学差异。siRNA转染使c-Fos表达减弱后,VSMC中α-SM-actin mRNA和蛋白表达较正常对照组及阴性siRNA组显著上调,bFGF、ET-1表达减弱,VSMC迁移速度减慢。结论 RNA干扰介导的c-Fos基因沉寂可在诱导VSMC分化的同时抑制VSMC迁移。Objective To investigate the influence of RNA interference targeting c-Fos gene on the phenotypic transition and migration of vascular smooth muscle cells（VSMCs）.Methods VSMCs were transfected with siRNA targeting c-Fos gene by liposome.Effects of c-Fos siRNA on mRNA expression of c-Fos and α-SM-actin were examined by RT-PCR while the levels of α-SM-actin,bFGF and ET-1 were determined by Western blot.We observed the differentiation of VSMCs by immunofluorescence and examined the migration ability of VSMCs with a modified Boyden-chamber assay simultaneously.Results After transfected with siRNA targeting c-Fos gene,expression of c-Fos was decreased compared to the control groups（normal control and negative siRNA groups）.The expression and mRNA level of α-SM-actin were increased in the c-Fos siRNA-transfected VSMCs.The shape of c-Fos siRNA-transfected VSMCs turned into spindle-like.c-Fos siRNA slowered VSMC migration and reduced the number of migrating VSMCs.Expressions of bFGF and ET-1 were significantly decreased after c-Fos siRNA transfection.Conclusion c-Fos gene silence through RNA interference can induce the phenotypic transition of VSMCs and inhibit their migration.

关 键 词：C-FOS RNA干扰 血管平滑肌细胞 细胞表型 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]