重组人骨形态发生蛋白-2和-6对人骨肉瘤细胞株MG63和U2OS的作用  

Effects of Recombinant Human Bone Morphogenetic Protein-2 and -6 on Human Osteosarcoma Cell Lines MG63 and U2OS

作　　者：何焕玲[1,2] 李星星[1,2] 吴丽美[1,2] 陈英华[1,2] 卫佳[1,2] 苗静琨[1,2] 周兰[1,2] 

机构地区：[1]重庆医科大学生物医学检验系 [2]临床检验诊断学教育部重点实验室,重庆400016

出　　处：《科技导报》2009年第17期47-51,共5页Science & Technology Review

基　　金：重庆市自然科学基金项目(CSCT2005BB5320)

摘　　要：为研究重组人骨形态发生蛋白rhBMP-2和rhBMP-6对人骨肉瘤细胞株MG63和U2OS的作用,分别用含rhBMP-2和rhBMP-6及重组绿色荧光蛋白的条件培养液干预人骨肉瘤细胞MG63和U2OS,利用台盼蓝拒染法、TUNEL法、吖啶橙/溴乙啶(AO/EB)荧光双染法、Transwell小室和碱性磷酸酶活性测定法分别检测细胞增殖、凋亡、迁移以及成骨分化能力的变化。结果显示与未实施任何干预的空白对照组和用rGFP干预实验对照组相比,rhBMP-2和rhBMP-6的干预致两种骨肉瘤细胞株:细胞存活率均随时间逐渐降低;凋亡率呈时间依赖性增加;穿膜数明显减低;碱性磷酸酶(ALP)活性逐渐增加。以上差异均有统计学意义(P<0.01)。表明rhBMP-2和rhBMP-6可抑制人骨肉瘤细胞株MG63和U2OS的增殖和迁移、诱导其凋亡和向成骨细胞分化。To investigate the effects of recombinant human BMP （rhBMP）-2 and -6 on osteosarcoma in treating human osteosarcoma cell lines MG63 and U2OS with rhBMP-2 and rhBMP-6, the osteosarcoma cells in experimental groups were treated with rhBMPs-CMs, those in control groups with rGFP-CM, and those in blank groups with neither. Cell proliferation, apoptosis transmigration and differentiation were detected at different times by Trypan blue exclusion test, transwell-room, terminal deoynucleotidyl transferase mediated dUTP nick end labeling （TUNEL）/acridine orange-ethidium bromide （AO/EB） double fluorescent dye staining and the alkaline phosphatase （ALP） activity, respectively. The main resuhs are as follows. （1） There are no significant differences of all indexes tested between the control groups and the blank （all with P〉 0.05）. （2） Compared with the control groups and the blank groups, the cell survival rates see a significant decrease when being treated with rhBMP-2 and rhBMP -6 （P〈0.01）. （3） The apoptosis indexes are increased （P〈0.01） and the results from TUNEL and AO/EB method are consistent. （4） The cell numbers of transmembrane are significantly decreased in both MG63 and U2OS （P〈0.01）. （5） The ALP activity is increased after being treated with rhBMP-2 and rhBMP-6 for 3 days in MG63 and for 5 days in U2OS （P〈0.01）. It is suggested that rhBMP-2 and -6 could inhibit the proliferation and transmigration of MG63 and U2OS, induce their apoptosis and differentiation.

关 键 词：骨形态发生蛋白 细胞增殖 细胞凋亡 细胞迁移 碱性磷酸酶活性 

分 类 号：R738.1[医药卫生—肿瘤]