过表达牙本质涎磷蛋白的骨髓间充质干细胞部分牙向和骨向分化基因的表达  

Expression of Odontogenic and Osteogenic Genes in Bone Marrow Mesenchymal Stem Cells with Overexpression of Mouse Dentin Sialophosphoprotein

作　　者：朱锋[1] 聂蓉蓉[2] 吴凌[1] 刘磊[1] 汤炜[1] 田卫东[1] 

机构地区：[1]四川大学华西口腔医院口腔疾病研究国家重点实验室 [2]南京大学医学院附属口腔医院修复科

出　　处：《四川大学学报（医学版）》2008年第2期290-293,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30471904);中国博士后科学基金(20060391019);教育部重点研究项目(106134)资助

摘　　要：目的观察小鼠骨髓间充质干细胞(BM-MSC)过表达牙本质涎磷蛋白(DSPP)后的形态学改变,并研究其部分牙向及骨向分化基因的表达情况。方法体外培养小鼠BM-MSC,通过腺病毒介导的方式将DSPP基因转染小鼠BM-MSC,通过标记基因GFP检测转染效率,观察转染后细胞的形态学改变,RT-PCR方法检测过表达DSPP的BM-MSC有无成牙和成骨基因的mRNA表达。结果成功获得小鼠BM-MSC,经腺病毒介导的DSPP基因转染BM-MSC的转染效率可达42.7%,转染后的细胞出现分化,似成牙本质样细胞。转染后细胞表达DSPP、DMP1、Msx1/2、Pax9、Lhx6/7、Sox9、Cbfα1、Osx、Col等多种牙向和骨向发育分化基因。结论过表达DSPP的BM-MSC出现细胞分化,并可表达多种牙向及骨向发育分化的基因。Objective To observe the overexpression of dentin sialophosphoprotein （DSPP） in mouse bone marrow mesenchymal stem cells （BM-MSC） and the expression of specific gene involved in odontogenic and osteogenic differentiation in transfected cells. Methods Mouse bone marrow mesenchymal stem cells （BM-MSC） were cultured and then transfected with DSPP gene by adenovirus-mediated way in vitro. The transfection efficiency of DSPP gene was assessed by the marker gene green fluorescent protein（GFP）. With having observed the morphological transformation of transfected BM-MSC, we studied whether the transfected BM-MSC would express odontogenic and osteogenic genes by RT-PCR. Results The mouse BM-MSC were obtained and the adenovirus mediated DSPP gene used to transfect BM-MSC successfully. The transfected efficiency was 42. 7%. The transfected BM-MSC were induced to differentiate into odontoblast-like cells and meanwhile expressed specific odontogenic and osteogenic differentiation genes such as DSPP , DMP1, Msx1/2, Pax9, Lhx6/7 , Sox9, Cbfα1 , Osx, Col Ⅰ. Conclusion The bone marrow mesenchymal stem cells can make differentiation with the overexpression of dentin sialophosphoprotein and express specific odontogenic and osteogenic differentiation genes.

关 键 词：牙本质涎磷蛋白 骨髓间充质干细胞 腺病毒 牙向分化 

分 类 号：Q786[生物学—分子生物学]