Nesprin-1在C_2C_(12)肌原细胞系分化过程中的作用  

The effect of Nesprin-1 on C_2C_(12) myoblast differentiation

作　　者：高放[1] 易岂建[1] 李晨[2] 田运娇[1] 王芳洁[1] Catherine M.Shanahan 张秋萍[2] 

机构地区：[1]重庆医科大学附属儿童医院心脏中心,重庆400014 [2]伦敦大学国王学院心血管研究室

出　　处：《复旦学报（医学版）》2010年第6期647-651,共5页Fudan University Journal of Medical Sciences

基　　金：高等学校博士学科点专项科研基金项目(20050631005);留学回国人员科研启动基金项目(教外司留[2007]24号文)

摘　　要：目的探讨Nesprin-1在肌细胞分化过程中的作用。方法将C2C12肌原细胞分化成肌小管,采用Western blot和免疫荧光技术检测Nesprin-1蛋白表达水平和亚细胞定位在肌小管分化成熟过程中的改变;同时利用RNA干扰技术抑制Nesprin-1蛋白的表达,观察其对C2C12肌原细胞分化的影响。结果 Nesprin-1各亚型在C2C12肌原细胞系分化过程中表达水平以及在细胞中的定位有明显改变。Nesprin-1 siRNA组与对照组相比,分化前后C2C12细胞中Nesprin-1蛋白表达明显降低,同时分化后肌组织分化成熟的蛋白标志肌球蛋白(myosin)的表达量以及肌小节的数量比对照组均明显减少(P<0.001)。结论本文证实Nesprin-1了对骨胳肌细胞分化至关重要,不同分子大小Nesprin-1亚型在C2C12肌原细胞分化过程中发挥各自的特殊作用,在该细胞质与细胞核中的也有不同的作用。Objective To investigate the role of Nesprin-1,a novel spectrin repeat containing scaffolding protein,in skeletal muscle cell differentiation.Methods During in vitro differentiation of C2C12 myoblasts into myotubes,changes in Nesprin-1 gene expression and sub-cellular localization were detected using Western blotting and immunofluorescence.RNA interference was used to reduce Nesprin-1 protein levels to further investigate the importance of Nesprin-1 in C2C12 myoblast differentiation.Results The Nesprin-1 isoform profile was altered in C2C12 myoblast differentiation.In addition,subcelluar fractionation revealed changes in subcellular compartmentalization of Nesprin-1.Furthermore,compared with the control group,myotube formation was dramatically reduced after Nesprin-1 depletion（P0.001）.Conclusions This is the first report to show that Nesprin-1 plays important roles in skeletal muscle cell differentiation.Nesprin-1 isoforms may perform specific functions during this process of C2C12 myoblast differentiation as well as have different roles in the cytoplasm and the nucleus of these cells.

关 键 词：Nesprin-1 C2C12肌原细胞系 分化 RNA干扰 

分 类 号：Q343[生物学—遗传学]