顺式调控元件NF-κB在尼古丁诱导ICAM-1表达中的作用  

NF-κB site of promotor mediates nicotine-induced ICAM-1 expression in human umbilical vein endothelial cells

作　　者：周卫辉[1] 陈祥银[1] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所病理生理室,北京100005

出　　处：《中国病理生理杂志》2003年第2期145-149,共5页Chinese Journal of Pathophysiology

基　　金："九五"国家科技攻关项目 (No .96 - 90 6 - 0 2 - 0 3)

摘　　要：目的 :研究 5’上游序列在尼古丁诱导细胞间粘附分子 - 1 (ICAM - 1 )基因转录调控中的作用。方法 :首先采用分子克隆技术构建实验所需的DNA片段 ,然后采用真核细胞转染技术将重组DNA片段与内参照质粒一起转染体外培养的内皮细胞并检测尼古丁刺激的转染细胞荧光素酶的相对活性。结果 :成功构建了 6种含有不同长度ICAM - 1基因启动子序列以及启动子中NF -κB和Sp - 1位点突变序列的荧光素酶报告基因质粒。与对照组相比尼古丁可促进含有启动子序列 - 579bp(pGL3E - 579/ +36) (P <0 .0 5) ,- 2 30bp (pGL3E - 2 30 / +36) (P<0 .0 5)以及启动子Sp- 1位点突变体 (pGL3E -Sp - 1 -MU) (P <0 .0 5)的荧光素酶基因表达 ,而下游NF -κB位点的删切重组体 (pGL3E - 1 34/ +36)和含有NF -κB位点突变体 (pGL3E -NF -κB -MU)的重组荧光素酶报告基因质粒在尼古丁诱导下与对照组相比荧光素酶表达量无明显差异。结论 :ICAM - 1基因启动子顺式作用元件下游NF -κB位点在尼古丁诱导ICAM -AIM: To study the mechanisms of nicotine-induced expression of intercellular adhesion molecule-1(ICAM-1). METHODS: Related luciferase reporter gene plasmids were constructed with molecular cloning techniques;above plasmids and intracontrol plasmid pSV-β-gal were co-transfected into human umbilical vein endothelial cells(HUVECs) with eukaryotic gene transfection techniques; the relative luciferase activities were detected in the transfected HUVECs. RESULTS: Series of luciferase reporter gene containing different sequences of human ICAM-1 promotor and site-directed mutants of NF-κB and Sp-1 in promotor were successfully constructed; Nicotine could increase the expression of luciferase reporter gene plasmid containing-579 bp(pGL3E-579/+36),-230 bp(pGL3E-230/+36) and mutated Sp-1 version(pGL3E-Sp-1-MU)( P< 0.05 vs control) of ICAM-1 promotor in the transfected HUVECs, whereas deletion derivative (pGL3E-134/+36) and mutation (pGL3E- NF-κB -MU) of downstream NF-κB site of ICAM-1 promotor prevent nicotine-induced increase in expression of luciferase reporter gene plasmid. CONCLUSION: NF-κB site of promotor mediates nicotine-induced ICAM-1 expression in human umbilical vein endothelial cells.

关 键 词：烟碱 NF-ΚB 细胞粘着因子 

分 类 号：R971.91[医药卫生—药品] R392.11[医药卫生—药学]