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骨形成蛋白-2基因转染对成纤维细胞表型影响的实验研究
引用本文:王磊,刘少华,张梅,曲迅,魏奉才. 骨形成蛋白-2基因转染对成纤维细胞表型影响的实验研究[J]. 华西口腔医学杂志, 2006, 24(3): 257-260
作者姓名:王磊  刘少华  张梅  曲迅  魏奉才
作者单位:山东大学齐鲁医院,口腔科,山东,济南,250012;山东医学高等专科学校,医学学科,山东,济南,250002;山东大学齐鲁医院,临床基础研究所,山东,济南,250012
基金项目:国家高技术研究发展计划(863计划)
摘    要:目的 探讨腺病毒载体介导的骨形成蛋白-2(Ad-BMP-2)基因转染人胚肺成纤维细胞(HELFs)后,对HELFs的表型特别是矿化能力的影响。方法 原代培养HELFs,并用Ad-BMP-2进行转染。观察转染前后HELFs的形态学变化;用四唑盐比色法、碱性磷酸酶染色、蛋白印迹以及体外矿化实验,检测转染BMP-2基因后HELFs增殖活性、碱性磷酸酶活性、BMP-2蛋白表达和矿化能力的变化。结果 HELFs经Ad-BMP-2转染后,胞体变大,由单一细长梭形转变为多种形态,由单层生长变为复层生长;转染后HELFs增殖受到明显抑制;碱性磷酸酶染色见大部分细胞呈现阳性反应;蛋白印迹检测到明显的BMP-2蛋白条带;细胞经茜素红S染色后出现桔红色的矿化结节。结论 经Ad-BMP-2转染后,HELFs向成骨细胞表型转变,具有了矿化能力。

关 键 词:骨形成蛋白-2  腺病毒载体  人胚肺成纤维细胞
文章编号:1000-1182(2006)03-0257-04
收稿时间:2005-01-18
修稿时间:2005-06-07

Experimental Study on the Phenotype of Human Embryonic Lung Fibroblasts Transfected with Ad-BMP-2
WANG Lei,LIU Shao-hua,ZHANG Mei,QU Xun,WEI Feng-cai. Experimental Study on the Phenotype of Human Embryonic Lung Fibroblasts Transfected with Ad-BMP-2[J]. West China journal of stomatology, 2006, 24(3): 257-260
Authors:WANG Lei  LIU Shao-hua  ZHANG Mei  QU Xun  WEI Feng-cai
Affiliation:1. Dept. of Dentistry, Qilu Hospital of Shandong University, Jinan 250012,China; 2. Dept. of Medicine, Shandong Medical College, Jinan 250002, China; 3. Basic Research Institute of Clinical Medicine, Qilu Hospital, Shandong University, Jinan 250012, China
Abstract:Objective To investigate the effects on the phenotype,especially the mineralization ability of human embryonic lung fibroblasts(HELFs) by transfection with adenovirus vector ecoding human bone morphogenetic protein-2 gene(Ad-BMP-2).Methods The HELFs were primarily cultured,then transfected with Ad-BMP-2.The morphologic characteristics of the cells were observed.The cell proliferation,alkaline phosphatase activities,BMP-2 protein expression,and the mineralization ability were detected with the methods of MTT,ALP staining,Western blot,and alizarin red S staining,respectively.Results After transfection,the shape of HELFs changed from silm spindle to multifigure,the cells became bigger than before.The colonies changed from unilaminar into multilaminar.The proliferation of HELFs was severely inhibited after transfection.An obvious BMP-2 lane was shown in Western blotting.Most cells presented positive in ALP staining,and large number of nacarat mineralized nodes were observed after alizarin red S staining.Conclusion HELFs were capable of transforming into osteoblast-like phenotype,and were endowed with the ability of mineralization while being transfected with Ad-BMP-2.
Keywords:bone morphogenetic protein-2   edenovirus vector   human embryonic lung fibroblasts
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