Abstract: Objective Construct a recombinant adenovirus containing the human hydroperoxidase(hTPO)gene,transfer the hTPO genes and human sodium/iodide symporter(hNIS)gene into glioma cells and study the iodide uptake ability of the cells and the killing effect of radioiodine,so as to investigate the feasibility of radioiodine therapy on glioma.Methods The recombinant adenovirus AdTPO was constructed by AdEasy system.Tranfecting hNIS gene into human glioma cell line U251 through recombinant plasmid,the stably expressing hNIS gene cell line hNIS-U251 was determined as negative control group.Then hTPO gene was transfected into hNIS-U251 by recombinant adenovirus AdTPO,achieving AdTPO-hNIS-U251,determined as testing group.U251 cell line without hNIS or hTPO gene was applied as control group.We investigated the iodide uptake assay,perchlorate suppressive assay,iodide organification assay to confirm gene expression and function.Killing effect of radioiodide on tumor cells was studied by clonogenic assay in vitro.Results Recombinant adenovirus AdTPO was correctly constructed.we were successful in co-transfecting hTPO gene into stably expressing hNIS cell line(hNIS-U251),obtaining AdTPO-hNIS-U251.In hNIS-U251 group,the iodide uptake ability(55769.96±4353.26count·min^-1)was 110 folds higher than that in control group(507.67±57.69count·min^-1),the effective half-life time was 7 minutes,the organification percentage was 0.1%,and the rate of clonal forming was(9.08±2.86)%,that was 10 folds lower compared with control group.While in AdTPO-hNIS-U251 group,the iodide uptake ability(74647.53±3605.88count·min^-1)was 147 folds higher than that in control group,the effective half-life time prolonged to 13 minutes,the organification percentage reached to 10%,and the rate of clonal forming was(6.80±2.09)%,which was 13 folds lower than control group respectively.Conclusion By co-transfection of hTPO and hNIS genes into glioma cell line U251,the iodide uptake ability increased markedly,hTPO increased the retension of iodide in the cells,and the ¹³¹I could kill tumor cells effectively.