转录因子 JunB 和 JunD 调控成釉细胞 MMP-20基因转录活性的研究
전록인자 JunB 화 JunD 조공성유세포 MMP-20기인전록활성적연구
Study of Transcription Factor JunB and JunD on Regulating the Expression of Gene MMP-20
  • 万方数据
    潍坊医学院学报 유방의학원학보
    JOURNAL OF WEIFANG MEDICAL COLLEGE
    2013年 1期 14-17 ,共4页

    郝佳*%张新新%袁杰%张娟娟%刘晓影%孙岩%高玉光

    학가*%장신신%원걸%장연연%류효영%손암%고옥광

    转录因子JunB,JunD%MMP20%双荧光素酶基因检测报告%基因定点突变 轉錄因子JunB,JunD%MMP20%雙熒光素酶基因檢測報告%基因定點突變
    전록인자JunB,JunD%MMP20%쌍형광소매기인검측보고%기인정점돌변
    JunB,JunD transcription factor%MMP20%Double luciferase genetic testing report%Gene point mutation
    目的通过研究转录因子JunB,JunD对成釉细胞中MMP-20基因表达的调控作用,从而进一步明确Jun家族在牙釉质形成中的影响.方法首先构建JunB和JunD真核表达载体重组质粒并瞬时转染重组质粒进入成釉细胞中;利用双荧光素酶基因检测报告系统分析不同浓度JunB,JunD对MMP-20启动子特征性序列区域的转录活性的影响;确定有明显作用的启动子区段,利用基因定点突变和双荧光素酶基因检测报告系统分析JunB对MMP-20基因启动子转录活性的影响;最后再观察JunB与JunD共转染时对MMP-20基因活性表达的改变.结果成功构建JunB,JunD真核重组表达载体,顺利将重组质粒转染入成釉细胞;双荧光素酶基因检测报告系统分析显示JunB对MMP20启动子活性表达上调,而JunD对MMP20启动子活性表达无作用;突变MMP20启动子AP1的两个结合位点后,MMP20启动子的转录活性下降,同时JunB也失去了上调MMP20启动子转录活性的作用.最后将JunB和JunD共转染时,在JunB存在的前提下,随着JunD转染量的增加,小鼠成釉细胞MMP20启动子转录活性明显减弱.结论该研究表明转录因子JunB与成釉细胞内MMP-20启动子的特征性序列相互作用,从而调控MMP20的表达水平;而JunD对MMP20启动子特征性序列无明显作用.所以为进一步研究Jun家族成员在釉质发育过程中的作用建立了重要的生物学基础.
    목적통과연구전록인자JunB,JunD대성유세포중MMP-20기인표체적조공작용,종이진일보명학Jun가족재아유질형성중적영향.방법수선구건JunB화JunD진핵표체재체중조질립병순시전염중조질립진입성유세포중;이용쌍형광소매기인검측보고계통분석불동농도JunB,JunD대MMP-20계동자특정성서렬구역적전록활성적영향;학정유명현작용적계동자구단,이용기인정점돌변화쌍형광소매기인검측보고계통분석JunB대MMP-20기인계동자전록활성적영향;최후재관찰JunB여JunD공전염시대MMP-20기인활성표체적개변.결과성공구건JunB,JunD진핵중조표체재체,순리장중조질립전염입성유세포;쌍형광소매기인검측보고계통분석현시JunB대MMP20계동자활성표체상조,이JunD대MMP20계동자활성표체무작용;돌변MMP20계동자AP1적량개결합위점후,MMP20계동자적전록활성하강,동시JunB야실거료상조MMP20계동자전록활성적작용.최후장JunB화JunD공전염시,재JunB존재적전제하,수착JunD전염량적증가,소서성유세포MMP20계동자전록활성명현감약.결론해연구표명전록인자JunB여성유세포내MMP-20계동자적특정성서렬상호작용,종이조공MMP20적표체수평;이JunD대MMP20계동자특정성서렬무명현작용.소이위진일보연구Jun가족성원재유질발육과정중적작용건립료중요적생물학기출.
    Objective Through the research of transcription factor JunB and JunD on regulating the expres-sion of gene MMP20 in ameloblast,thus further clearing the influence of Jun family in enamel development .Methods Firstly recombinant plasmid of eukaryotic expression vector of JunB and JunD was constructed and recombinant plasmid was transfected into ameloblast instantly .Double luciferase genetic testing report system were performed to measure the effect of different concentration of JunB and JunD to transcription activity of MMP 20 promoter.To make sure the promoter section of apparent effect,and to analyse the influence of JunB/D to MMP20 gene promoter transcription activity by using the gene point mutation and double luciferase genetic testing report system .Finally the changes of MMP20 gene expres-sion were observed when the JunB and JunD commonly transfect .Results Eukaryotic recombination expression vector of JunB and JunD were constructed successfully ,and the recombinant plasmid was transfect into ameloblast smoothly .Doub- @@@@le luciferase genetic testing report system analysis showed that JunB can rise the activity expression of MMP 20 promoter, bur JunD have no action to MMP20 promoter activity express.After mutating MMP20 promoter AP1 two binding site, transcription activity of MMP20 promoter declined,and at the same time JunB had lost its action to rise MMP20 promoter transcription activity.Finally When JunB and JunD commonly transfect into ameloblast ,JunB existed in the premise,with the increased dose of JunD transfection ,mice into glaze cell transcription activity of MMP20 promoter decreased signifi-cantly in mice ameloblast.Conclusion The study suggests that transcription factor JunB were interacted with the char-acteristic sequence of MMP20 promoter in ameloblast,so as to control MMP20 expression level.But transcription factor JunD have no influence on the characteristic sequence of MMP 20 promoter.Further study the function of the Jun family in the process of enamel development can establish the important biological basis .
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