293T细胞转染表达ARG1及其对砷代谢的影响

Expression of ARG1 Gene in 293T Cell and Its Influence on Arsenic Metabolism

研究含有ARGI基因的真核表达载体PcDNA3.1-IE-ARG1转染真核细胞后,在细胞中的表达及对细胞砷代谢的影响。由脂质体介导将PcDNA3.1-IE-ARG1转染入293T中,用real-timePCR方法检测ARG1基因的表达,用原子吸收分光光度法测定24h砷染毒后细胞内砷含量及细胞排砷量。重组质粒成功转入293T细胞中且表达良好;与转染前细胞比较,转染重组质粒的细胞在不同浓度砷染毒下细胞内砷含量明显降低(P<0.05),并且细胞的排砷量也高于转染前的;此外,转染表达ARGI基因的细胞在砷染毒48h后其内GSH及GST的含量高于转染前细胞。这些表明,ARG1基因在真核细胞中相对高表达后在砷代谢排出中发挥了一定的作用。

To examine the expression ofARG1 in eukaryotic cell and the effect of arsenic metabolism after transfecting the eukaryotic cell vector.293T cell was transfected with the recombinant plasmid and the expression of recombinant ARG1 was examined by real-timePCR.Cells were exposed to arsenite for 24h,Arsenite accumulation and efflux of cell was determined by atomic absorption spectrophotometry.The recombinant plasmid was successfully transfected into 293T cell.Arsenite contents in the transfected-cells that were exposed to arsenite with different consistency were lower than those in the 293T cell（P0.05）,and the efflux of cell was higher than those in the 293T cell.The transfected-cells were exposed to arsenite for 48h,intracellular concentration of GSH and GST activity were higher than those in the 293T cell.ARG1 plays an important role in arsenic metabolism after high expression of ARG1 in eukaryotic cell.