全反式维甲酸通过Sp1/Sp3位点调节HEK-293细胞中CD2AP基因的表达  

All-trans retinoic acid regulates CD2AP gene expression through Sp1/3-binding sites in HEK293 cells

作　　者：高珊 徐华国 金蕊 邱灵芝 朱亮华 蒋莉 庄丽丽 周国平 

机构地区：[1]南京医科大学第一附属医院儿科,江苏省210029

出　　处：《江苏医药》2013年第9期999-1002,共4页Jiangsu Medical Journal

基　　金：国家自然科学基金资助项目(81170661);国家自然科学基金资助项目(30872804);江苏高校优势学科建设工程资助项目(JX10231081);江苏省医学重点人才(RC2007050);教育部博士点基金(20113234110010);江苏省普通高校研究生科研创新计划项目(CX2211_0714)

摘　　要：目的研究全反式维甲酸(ATRA)对人胚胎肾HEK-293细胞中CD2相关蛋白(CD2AP)基因表达的影响。方法 ATRA作用HEK-293细胞后,采用RT-PCR和Western blot分别检测CD2AP mRNA及其蛋白的表达。将构建的不同Sp1/Sp3位点的定点突变、巢式缺失突变的CD2AP启动子质粒转染HEK-293细胞,检测报告质粒的荧光素酶活性。结果 ATRA明显上调HEK-293细胞中CD2AP mRNA和蛋白水平(P<0.05)。缺失突变及点突变研究显示,Sp1/Sp3-A、C位点在介导ATRA上调CD2AP过程中发挥重要作用。结论 ATRA通过Sp1/Sp3结合位点诱导HEK-293细胞中CD2AP的表达;ATRA对CD2AP基因缺陷肾病患者可能有治疗作用。Objective To investigate the effect of all-trans retinoic acid（ATRA） on the CD2- associated protein（CD2AP） gene expression in HEK293 cells. Methods After HEK-293 cells were treated with ATRA, the mRNA and protein expressions of CD2AP were detected by R-T-PCR and Western blot, respectively. Plasmids containing different lengths of deletion mutations of human CD2AP promoter were constructed and transfected into HEK-293 cells. Promoter activity was determined by luciferase analysis after treated with ATRA 20μM for 48 h. Results ATRA could significantly upregulate the mRNA and protein expressions of CD2AP in HEK-293 cells（P〈0. 05）. Progressive deletion and site-directed mutagenesis analysis indicated that ATRA increased CD2AP promoter activity through Sp1/Sp3-A, C binding sites. Conclusion ATRA induces CD2AP expression through Sp1/Sp3-binding sites in HEK-293 cells. ATRA may be effective in treating patients with CD2 AP-deficient.

关 键 词：全反式维甲酸 HEK-293细胞 CD2相关蛋白 

分 类 号：R725[医药卫生—儿科]