MEF2C调控成釉细胞MMP-20基因启动子转录活性的研究  

Transcription factor MEF2C regulates the promoter activity of MMP-20 gene in ameloblasts

作　　者：刘珩[1] 李伯翰[2] 韩婷婷[2] 李东亮[2] 王玉敏[2] 孙岩[1] 曲政[1] 高玉光[2] 

机构地区：[1]山东潍坊医学院口腔医学研究所,潍坊261042 [2]山东滨州医学院附属医院口腔内科,滨州256603

出　　处：《牙体牙髓牙周病学杂志》2013年第4期214-217,230,共5页Chinese Journal of Conservative Dentistry

基　　金：国家自然科学基金资助项目(30973327);山东省自然科学基金资助项目(ZR2010HM076)

摘　　要：目的:通过研究MEF2C转录因子对小鼠成釉细胞金属基质蛋白酶-20(matrix metalloprotei-nase-20,MMP-20)基因表达的调控作用,明确MEF2C的功能,为研究MEF2C在釉质形成中的作用奠定基础。方法:用RT-PCR的方法扩增小鼠MEF2C基因,并构建真核表达载体pcDNA3.1/myc-hisA-MEF2C;对MMP-20基因启动子区域(-1197～+23)中MEF2C潜在的结合位点进行定点突变,并构建至荧光报告载体pGL3-Basic中;利用双荧光素酶基因报告系统分析MEF2C对MMP-20基因启动子转录活性的影响。结果:在成釉细胞中过量表达MEF2C能够明显提高MMP-20基因启动子的转录活性,而对定点突变后的MMP-20基因启动子转录调控作用减弱。结论:小鼠成釉细胞核中MEF2C可通过MMP-20启动子上MEF2C潜在的结合位点,上调MMP-20基因表达水平。AIM ： To study the function of transcription factor MEF2C in the regulation the of matrix metal- loproteinase-20 （MMP-20） expression in ameloblasts. METHODS： The eukaryotic expression vector peDNA3.1/ myc-HisA-MEF2C was constructed through amplification of mouse MEF2C gene by RT-PCR. MEF2C possible binding sites were site-directed for mutagenesis in MMP-20 gene promoter region （ - 1197 N ＋ 23 ） with a fluorescent reporter vector pGL3-Basic. Transiently transfection was finished in mouse ameloblasts. The effect of MEF2C on transcriptional activity of MMP-20 was detected by dual-luciferase reporter assay. RESULTS ： Transcriptional activity of the MMP - 20 gene promoter（ -1197 to ＋ 23） in mouse ameloblasts was significantly promoted by over-expression of MEF2C, while the transcriptional activity of the MMP-20 gene promoter（ -1197 to ＋ 23 ） was reduced by site-directed muta- genesis. CONCLUSION： MEF2C up-regulates MMP-20 gene expression via the combination with the MEF2C bind- ing sites of MMP-20 gene promoter in ameloblasts.

关 键 词：转录因子MEF2C 基质金属蛋白酶-20 成釉细胞 定点突变 

分 类 号：R780.2[医药卫生—口腔医学]