Zhang Yi-ping, Qin Feng, Wu Chang-jing, Li You-rui, Chen Rui, Shao Min-feng, Fu Qiang
Department of Prosthodontics, Guanghua School of Stomatology, Sun Yat-Sen University, Guangzhou 510055, China.
Zhonghua Yi Xue Za Zhi. 2009 Dec 1;89(44):3143-6.
To study the effects of cytoskeleton reorganization inhibition with LIMK2 RNAi upon the mechanosensitivity of c-fos gene in osteoblast.
Mouse primary osteoblast was treated with LIMK2 specific siRNA (RNAi Group), negative control siRNA (NC Group), and then were loaded or unloaded by fluid shear stress. Real-time PCR and immunofluorescence were used to detect the c-fos expression levels and statistics analysis was performed.
When the cytoskeleton reorganization was inhibited with RNAi only, the c-fos mRNA (0.0108 +/- 0.0074 and 0.0042 +/- 0.0018, t = -1.86, P > 0.05) and protein (121 +/- 7 and 119 +/- 6, t = -1.272, P > 0.05) expression levels of each unloaded group had no significant difference; Fluid shear stress could up-regulate the c-fos mRNA (0.2203 +/- 0.1532 vs 0.0042 +/- 0.0018, t = -707.35, P < 0.05)and protein (178 +/- 12 vs 119 +/- 6, t = -30.761, P < 0.05) expression; After the cytoskeleton reorganization was inhibited with RNAi, the c-fos mRNA (0.5280 +/- 0.0879 vs 0.2203 +/- 0.1532, t = -1007.00, P < 0.05) and protein (224 +/- 46 vs 178 +/- 12, t = -6.853, P < 0.05) expression induced by fluid shear stress had significant difference. Cytoskeleton reorganization inhibition with RNAi had synergistic effect upon the expression of c-fos mRNA (F = 84.388, P < 0.05) and protein (F = 42.409, P < 0.05) induced by fluid shear stress.
Using RNAi against LIMK2 to inhibit the cytoskeleton reorganization can promote the expression of c-fos gene and thus enhance the mechanosensitivity of c-fos gene in osteoblast.
研究采用LIMK2 RNA干扰抑制细胞骨架重排对成骨细胞中c-fos基因机械敏感性的影响。
用LIMK2特异性小干扰RNA（RNA干扰组）、阴性对照小干扰RNA（阴性对照组）处理小鼠原代成骨细胞，然后对其施加或不施加流体剪切力。采用实时荧光定量PCR和免疫荧光法检测c-fos表达水平并进行统计学分析。
单纯采用RNA干扰抑制细胞骨架重排时，各未加载组的c-fos mRNA（0.0108±0.0074和0.0042±0.0018，t=-1.86，P>0.05）和蛋白（121±7和119±6，t=-1.272，P>0.05）表达水平无显著差异；流体剪切力可上调c-fos mRNA（0.2203±0.1532比0.0042±0.0018，t=-707.35，P<0.05）和蛋白（178±12比119±6，t=-30.761，P<0.05）表达；采用RNA干扰抑制细胞骨架重排后，流体剪切力诱导的c-fos mRNA（0.5280±0.0879比0.2203±0.1532，t=-1007.00，P<0.05）和蛋白（224±46比178±12，t=-6.853，P<0.05）表达有显著差异。采用RNA干扰抑制细胞骨架重排对流体剪切力诱导的c-fos mRNA（F=84.388，P<0.05）和蛋白（F=42.409，P<0.05）表达有协同作用。
利用针对LIMK2的RNA干扰抑制细胞骨架重排可促进c-fos基因表达，从而增强成骨细胞中c-fos基因的机械敏感性。
