E-box元件在p21基因表达调控中作用的研究  

Study on effect of the element of E-box in regulating the expression of p21 gene

作　　者：孙荣距[1] 夏照帆[2] 杨宗城[3] 姚咏明[1] 赵晓东[1] 苏踊跃[3] 罗向东[3] 

机构地区：[1]解放军总医院第一附属医院急救部,北京100037 [2]第二军医大学附属长海医院烧伤研究所,上海200433 [3]第三军医大学附属西南医院烧伤研究所,重庆400038

出　　处：《感染．炎症．修复》2008年第2期90-93,共4页Infection Inflammation Repair

基　　金：国家自然科学基金重点项目(10332060);国家自然科学基金资助项目(30700869)

摘　　要：目的:通过p21基因调控序列E-box位点的突变分析,研究E-box位点对p21基因表达调控的作用。方法:将人野生型p21基因启动子全长序列构建到荧光素酶报告基因载体pGL3-basic,并进一步对重组载体pGL3-p21p启动子E-box位点做定点突变,构建突变载体E1、E2、E3,分别转染人脐静脉血管内皮细胞株(ECV304细胞),DLR系统测定荧光素酶活性,并观察E-box位点突变前后荧光素酶活性变化。结果:测序结果表明野生型pGL3-p21p及突变载体E1、E2、E3构建成功。野生型pGL3-p21p荧光素酶活性(105.82±16.55)明显高于空载体pGL3-basic(1.56±0.62);突变载体E1、E2、E3的荧光素酶活性(70.71±16.46、37.87±3.26、83.25±11.48)较野生型载体不同程度下降(P<0.01)。结论:p21基因启动子某些E-box位点参与了p21基因的表达调控。Objective：To study point mutation of E-box of p21 gene promoter and its effect on expression of p21 gene. Methods：The wild-type human p21 gene promoter was subeloned into pGL3-basic plasrnid, and mutant vectors of E1 ,E2 and E3 were reconstructed by making point mutations of E-box elements of p21 gene promoter, and all the above vectors were transfected into ECV304 cells. The dual luciferase activities were assayed by DLR system. Results： Wild-type pGL3-p21p and mutant vectors were constructed successfully, and luciferase expression induced by pGL3-p21p （105. 82±16. 55） was much higher than that by pGL3-basie, while luciferase activity induced by mutant vectors E1 （70. 71±16.46）, E2（37.87±3.26）, E3（83.25±11.48） was declined at extent after being transfected into cells. Conclusion： Some E-box elements of promoter might be involved in the expression of p21 gene.

关 键 词：P21 E-box元件 报告基因 

分 类 号：R730.2[医药卫生—肿瘤] R735.202[医药卫生—临床医学]