沉默LIMK2基因对成骨细胞c—fos基因力学敏感性的影响  

Effects of LIMK2 RNA interference on the mechanosensitivity of c-fos gene in osteoblast

作　　者：张艺平[1] 覃峰[1] 吴昌敬 李友瑞[1] 陈睿[1] 邵敏峰[1] 付强[1] 

机构地区：[1]中山大学光华口腔医学院附属口腔医院修复科中山大学口腔医学研究所,广州510055 [2]广州市中西医结合医院口腔科

出　　处：《中华医学杂志》2009年第44期3143-3146,共4页National Medical Journal of China

基　　金：基金项目：广东省科技计划基金（20088030301121、20098050700027）;广东省医学科研基金（A2008227）

摘　　要：目的采用RNA干扰沉默LIMK2基因来抑制细胞骨架的改建，观察流体剪切力作用下其对成骨细胞c—fos基因力学敏感性的影响。方法对小鼠颅骨分离的成骨细胞分别给予RNA干扰、阴性RNA干扰2种处理后，进行流体剪切力加载或不加载。分别应用逆转录（RT）一PCR和免疫荧光检测c—fosm RNA和蛋白的表达，并进行统计学分析。结果在无流体剪切力加载条件下，单纯应用RNA干扰方法抑制细胞骨架改建，并不能使成骨细胞c-fos mRNA（0．0108±0．0074与0．0042±0．0018，t=-1．86，P〉0．05）和蛋白（121±7与1194-6，t=-1．272，P〉0．05）的表达升高；流体剪切力能使成骨细胞c-fosmRNA（0．22034-0．1532比0．0042±0．0018，t=-707．35，P〈0．05）和蛋白（1784-12比119±6，t=-30．761，P〈0．05）的表达水平显著升高；RNA干扰处理可显著提高流体剪切力诱导成骨细胞c-fosmRNA（0．52804-0．0879比0．22034-0．1532，t=-1007．00，P〈0．05）和蛋白（224±46比1784-12，t=-6．853，P〈0．05）的表达水平；采用RNA干扰的方法抑制细胞骨架的改建，可以对流体剪切力诱导成骨细胞c—fosmRNA（F=84．388，P〈0．05）和蛋白（F=42．409，P〈0．05）的表达起协同作用。结论采用RNA干扰沉默LIMK2基因抑制细胞骨架的改建，可以促进成骨细胞在流体剪切力作用下的c-fos基因表达。Objeelive To study the effects of cytoskeleton reorganization inhibition with LIMK2 RNAi upon the mechanosensitivity of c-fos gene in osteoblast. Methods 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 immunofluoreseence were used to detect the c- fos expression levels and statistics analysis was performed. Results 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 cotdd up-regulate the c-los 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 e-los 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. Conclusion Using RNAi against LIMK2 to inhibit the cytoskeleton reorganization can promote the expression of c-fos gene and thus enhance the meehanosensitivity of c-fos gene in osteoblast.

关 键 词：成骨细胞 基因 FOS 力学 流体剪切力 

分 类 号：R686[医药卫生—骨科学]