人成骨肉瘤MG-63细胞分化特性及分化过程中的基因表达  

Differentiation and gene expression of human osteosarcoma cell line MG-63

作　　者：罗湘杭[1] 廖二元[1] 周后德[1] 伍贤平[1] 

机构地区：[1]中南大学湘雅二医院代谢内分泌研究所,长沙410011

出　　处：《湖南医科大学学报》2001年第2期107-110,共4页Bulletin of Hunan Medical University

基　　金：国家"九五"攻关项目!( 96 90 6 0 5 0 5 );国家自然科学基金!( 3 9970 3 4 7)

摘　　要：观察人成骨肉瘤MG 6 3细胞分化特性及分化过程中的基因表达。在细胞培养的不同时间 ,用α 磷酸奈酚法测定碱性磷酸酶 (ALP)活性 ;放射免疫法测定骨钙素 (BGP)含量 ;半定量逆转录聚合酶链反应测I型胶原、基质金属蛋白酶 (MMP) 1和基质金属蛋白酶抑制因子 (TIMP) 1基因mRNA表达 ;VanGieSon氏苦味酸酸性复红染色法染色细胞I型胶原。结果表明MG 6 3细胞接种培养第 7d后I型胶原基因表达较高。MMP 1表达量随时间推移逐渐增加 ,至第 2 4d达到高峰。第 1～ 9dTIMP 1表达量逐渐增加 ,其后基本恒定。ALP活性第 0～ 12d逐渐增高 ,至第 12d达最高 ,其后逐渐下降。第 18d后 ,细胞有许多大小不等的结节形成 ,I型胶原结节染色较无结节处深。MG 6 3细胞具有成骨细胞表型特征。MG 6 3细胞生长分为细胞增殖、骨基质成熟、骨基质矿化阶段 ,且I型胶原 ,MMP 1,TIMP 1基因表达及ALP活性呈时间特异性。To observe the differentiation and gene expression of human osteosarcoma cell line MG 63 in culture. Alkaline phosphatase (ALP) activity was determined by p nitrophenyl phosphate assay; bone Gla protein (BGP) was measured by radioimmunoassay; type I collagen, matrix metalloproteinase (MMP) 1, tissue inhibitor of matrix metalloproteinase (TIMP) 1 mRNA were examined using semiquantitive reverse transcription polymerase chain reaction (RT PCR) analysis; MG 63 cells were stained by the Van GieSon method. Type I collagen mRNA expression achieved a maximum level on the 17th day in MG 63 cells; MMP 1 mRNA was not expressed until the 5th day of culture, and gradually increased; TIMP 1 mRNA was nearly constant; ALP activity gradually up regulated during 0～12 days, and decreased on the 18th day. By Van GieSon staining, MG 63 cells displayed nodule formation at the 12th day, and became more prominent on the 18th day. The results indicate that human osteosarcoma cell line MG 63 has the osteoblast phenotype; during the differentiation of MG 63 cells , there are the following three principle periods: proliferation, extracellular matrix maturation and mineralization.

关 键 词：MG-63细胞 细胞分化 骨肿瘤 基因表达 人成骨肉瘤 

分 类 号：R73-3[医药卫生—肿瘤]