重组Nox1在NIH3T3细胞中的表达及其对活性氧的影响  

Expression of Recombinant Nox1 in NIH3T3 Cells and its Impact on Reactive Oxygen Species

作　　者：仓辉[1] 邹嫣琼[1] 杨洁[1] 王毓美[1] 王英[1] 史桂英[1] 易静[1] 

机构地区：[1]上海第二医科大学基础医学院细胞生物学教研室,上海200025

出　　处：《上海第二医科大学学报》2005年第5期471-474,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金(30170475);上海市教委第四期重点学科(ZDXK2001)资助项目.

摘　　要：目的构建Nox1的真核表达载体,探讨其在NIH3T3细胞中表达后对活性氧水平和药物细胞毒易感性的影响。方法构建携带Nox1基因的真核表达载体pcDNA3-Nox1,并将其转染NIH3T3细胞,采用RT-PCR方法检测Nox1在mRNA水平上的表达,检测细胞活性氧和细胞活力。结果RT-PCR显示转染后的NIH3T3细胞可以有效地转录Nox1mRNA,细胞中的活性氧水平明显升高,同时对As2O3细胞毒作用更加敏感。结论含有Nox1cDNA的重组质粒pcDNA3-Nox1转染细胞后可有效表达Nox1,使活性氧水平升高,并因此增加细胞对药物细胞毒的敏感性。Objective To construct Nox1 eukaryotic expressing vector and to explore its impact upon reactive oxygen species (ROS) and cytotoxic susceptibility after its expression in cells. Methods Nox1 eukaryotic expressing vector pcDNA3-Nox1 was constructed and transfected to NIH3T3 cells. Nox1 expression was detected with RT-PCR. The cellular ROS level and cell viability were evaluated. Results RT-PCR showed that transfected NIH3T3 could effectively transcript Nox1 mRNA. Cellular ROS increased remarkably and susceptibility to arsenic trioxide-induced cytotoxicity was enhanced. Conclusion NIH3T3 cells transfected with pcDNA3-Nox1 could express Nox1 effectively. Nox1 expression increased cellular ROS to some extent, which then enhanced cellular sensitivity to As 2O 3 accordingly.

关 键 词：NIH3T3细胞 活性氧 表达及 RT-PCR方法 真核表达载体 MRNA水平 细胞毒作用 AS2O3 细胞活力 转染细胞 重组质粒 cDNA 易感性 1基因 Nox 敏感性 药物 检测 升高 

分 类 号：Q786[生物学—分子生物学]