绿色荧光蛋白基因在脂肪细胞分化研究中的应用  

Application of green fluorescent protein gene in study of adipocyte differentiation

作　　者：祝骥[1,2] 马文丽[1] 毛向明[1] 李凌[1] 吴清华[1] 郑文岭[3] 

机构地区：[1]南方医科大学分子生物学研究所 [2]中国科学院华南植物研究所,广东广州510650 [3]广州军区广州总医院分子肿瘤研究所

出　　处：《第一军医大学学报》2005年第9期1119-1123,共5页Journal of First Military Medical University

基　　金：国家自然科学基金(39880032);广州市重点科技攻关项目(990448022)~~

摘　　要：目的建立人脂肪细胞分化的绿色荧光蛋白活体检测动态模型,为进一步进行脂肪细胞分化及其相关基因表达研究奠定基础。方法用PCR法从培养的人前脂肪细胞总DNA中扩增出过氧化物体酶增殖物激活受体γ2(PPARγ2)基因启动子,将该启动子插入真核细胞表达载体pEGFP-1后转染3T3成纤维细胞及人前脂肪细胞,并诱导脂肪细胞分化。结果在胰岛素、地塞米松及3-异丁基-1-甲基黄嘌呤诱导下,人前脂肪细胞分化为成熟脂肪细胞时表达绿色荧光蛋白基因,而3T3细胞则不能诱导为脂肪细胞及表达绿色荧光蛋白基因。结论脂肪组织特异表达的PPARγ2基因启动子与绿色荧光蛋白基因构成的重组基因转化人前脂肪细胞后,能够用来动态活体检测脂肪细胞分化状态。这一模型的建立为脂肪细胞分化调控的分子机制研究提供一个更加方便的检测途径。Objective To establish a model for dynamic detection of adipocyte differentiation in vivo by green fluorescent protein （GFP）. Methods The promoter of peroxisome proliferators-activated receptor gamma 2 （PPAR γ2） gene was amplified by PCR from the genomic DNA of human preadipocytes, inserted into the eukaryotic expression vector pEGFP-I and transferred into NIH3T3 fibroblasts and preadipocytes which were induced to differentiate into adipocyte. Results Upon induction with insulin, dexamethasone and 3-isobutyl-l-methyl xanthine, the transformed human preadipocytes differentiated into mature adipocytes and expressed GFP, but NIH3T3 fibroblasts could not differentiate into adipocytes or express GFP. Conclusion Transformation of human preadipocytes with the recombinant of PPARγ2 promoter with GFP gene can be employed to dynamically detect the differentiation of adipocytes in viro.

关 键 词：脂肪细胞分化 绿色荧光蛋白基因 过氧化物体酶增殖物激活受体γ2基因启动子 

分 类 号：R346[医药卫生—基础医学]