NOR_1基因对HepG_2细胞E-选择素的影响及机制探讨  

Effect of NOR1 Gene on Level of E-selectin in HepG_2 Cells and its Mechanism

作　　者：李跃进[1] 王巍巍[2] 李登清[3] 

机构地区：[1]南华大学第二附属医院检验科,421001 [2]南华大学第一附属医院检验科,421001 [3]中南大学湘雅医学院检验系,410013

出　　处：《实用癌症杂志》2010年第1期9-12,共4页The Practical Journal of Cancer

基　　金：国家自然科学基金资助项目(编号:30300383)

摘　　要：目的探讨NOR1基因对HepG2细胞E-选择素表达水平的影响及NF-kB通路是否参与了NOR1基因对HepG2的E-选择素的影响。方法将NOR1基因通过脂质体转染入HepG2细胞。分别设立HepG2细胞对照组、HepG2/pcDNA3.1(+)空白质粒细胞组、HepG2/pcDNA3.1(+)/NOR1转染阳性克隆细胞组、PDTC处理的HepG2/pcDNA3.1(+)/NOR1转染阳性克隆细胞组(后简称PDTC处理组),分别检测4组细胞E-选择素的mRNA水平和蛋白水平。结果基因芯片筛选转染NOR1基因的HepG2细胞E-选择素表达水平上调(cy3/cy5>2.0);RT-PCR结果显示转染阳性克隆细胞组E-选择素mRNA的表达水平明显高于空白组、对照组和PDTC处理组(P<0.01),ELISA结果显示转染阳性克隆细胞组E-选择素表达水平明显高于空白组、对照组和PDTC处理组(P<0.01)。结论NOR1基因诱导HepG2细胞E-选择素表达水平增高,NF-kB通路参与了NOR1基因对HepG2细胞E-选择素的影响。Objective To study the effect of NOR1 gene on level of E-selectin in HepG2 cells and NF-kB pathway on E- selectin expression in HepG2 transfeeted by NOR1. Methods NOR1 gene was introduced into human liver eancer cell line HepG2 by using cation lipofectamine. The expression of E-selectin mRNA in HepG2 cells was detected by reverse transcription polymerase chain reaction （RT-PCR） and fluorescence quantitive polymerase chain reaction （FQ-PCR） which were used to confirm the results of the DNA microarray. E-selectin in cell culture medium of NORI gene transfection group were measured by ELISA. Results The DNA microarray data suggested E-selectin was the up-regulated gene （cy3/cy5 〉 2.0）. The levels of E-selectin mRNA in NOR1 gene transfection group were higher than those in blank group, control group and PDTC group （P 〈 0.01 ） through RT-PCR and FQ-PCR; the levels of E-selectin in cell culture medium of NORI gene transfection group were higher than those in blank group,control group and PDTC group （P 〈 0.01）. Conclusion NOR1 gene may induce increased expression of E-selectin in HepG2 cells and NF-kB pathway maybe involved in this change.

关 键 词：E-选择素 NOR1基因 基因芯片 HEPG2细胞 实时荧光定量PCR 

分 类 号：R73-34[医药卫生—肿瘤]