摘要
目的构建含有胶原蛋白1A2启动子和增强子的人皮肤成纤维细胞(HDFs)特异的非病毒骨形成蛋白-2(BMP-2)表达载体pcDNA3-CEP-BMP-2,验证其是否增强BMP-2特异性在HDFs表达并增强成骨诱导作用,从而探索非病毒载体介导正常细胞的基因修饰用于骨修复的可能性。方法将胶原蛋白增强子、启动子及BMP-2基因导入pcDNA3质粒中,用脂质体法转入HDFs,将血管内皮细胞(VEC)作为对照,RT-PCR检测BMP-2基因在两种细胞中的表达差异,并检测转染后HDFs体外成骨表型。结果转染pcDNA3-CEP-BMP-2在HDFs造成BMP-2表达高于pcDNA3-BMP-2,而在VEC的BMP-2表达不增高。转染了pcDNA3-CEP-BMP-2的HDFs骨钙素的分泌和矿化能力高于pcDNA3-BMP-2。结论胶原蛋白1A2启动子和增强子能特异性地增强BMP-2基因在HDFs中的表达。
Objective To construct the fibroblast-specific non-viral vector pcDNA3-CEP-BMP-2 containing collagen 1A2 enhancer and promoter, and to validate the enhancement of BMP-2 expression in the human dermal fibroblasts by this vector, compared with the routine non-viral BMP-2 vector. Methods The sequences for collagen 1A2 enhancer and promotor, and BMP-2 gene were ligated into the pcDNA3 plasmids. The plasmids were transfected into human skin fibroblasts and vein endothelial cells by means of cationic liposomes. The expressions of the plasmids in these two kinds of cells were detected by RT-PCR. The osteogenic phonotypes of fibroblasts were determined. Results pcDNA3-CEP-BMP-2, which contained collagen 1A2 enhancer and promoter could enhance the BMP-2 expression in the fibroblasts but not in vein endothelial cells. Osteogenetic phenotypes were more obvious in the fibroblasts transfected with pcDNA3-CEP-BMP-2 than in pcDNA3-BMP-2-transfected ones. Conclusion Collagen 1A2 enhancer and promoter can enhance BMP-2 expression in fibroblasts.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2006年第5期466-469,共4页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(30170943)资助项目
