The Study Of Radioiodine Therapy Mediated By Transfer Of HNIS Gene Into Glioma Cell Controlled By HTERT Promoter In Vivo

Objective:Glioma is one of the worst of human survival prognosis tumors, with a high fatality rate and the characteristics of the low cure rate, the efficacy is not always ideal. A hot topic in current cancer research is gene therapy,131I treatment is a relatively efficient and safe method of treatment, we assume that banding the two methods together to treat glioma.131I for differentiated thyroid cancer with high efficiency and specificity of anti-effect, the molecular basis of its iodine uptake and storage of iodine is that the cell membrane contains these two encoded proteins of the human sodium iodide symporter (hNIS). We assume that these two protein coding genes cloned into glioma cells for specific expression, then radioactive iodine treatment.This study introduces human telomerase reverse transcriptase (hTERT) promoter. to built and apply recombinant adenovirus as a gene transfer vector. the hNIS gene’s transfection into glioma cells, while establishing glioma animalmodel, and further analysis of the hTERT’s targeted guiding hNIS-specific expression in glioma cells. to evaluate the131I ’s killing effects.Method:1Verify that the expression’s active of telomerase in glioma cells. hTERT core promoter was amplified by PCR; Constructing recombinant adenovirus carring hNIS gene activated by hTERT; weston-blot analysis the recombinant adenovirus2The establishment of U87cells in nude mice bearing tumor model, the overall level of hNIS and hNIS gene transfection mediated glioma radionuclides99mTcO4-imaging.31I whole body imaging, transplanted tumors and other organs’ intake of131I and131I’therapeutic effect on the transplanted tumor transFected hNIS gene in nude mice.Result:1Successfully constructed recombinant adenovirus Ad-hTERT-hNIS containing the hTERT promoter and the hNIS gene.2Successful establishment of U87cells in nude mice bearing tumor model, and the99mTcO4-imaging and131I scintigraphy of the transplanted tumors were positive, indicating that the gene transfection success, laying the foundation for further131I therapy.3Analysis radioactive counting of various organs of nude mice after the radionuclide131I imaging. senting tumor tissue to pathology for immunohistochemical analysis. the NIS protein positive.4Radionuclide therapy of gliomas. the experimental group compared with the tumor cell shrinkage, the median survival time of nude mice was significantly prolonged, a large patch of cellstissue necrosis in the tumors after iodine treatment for pathology, radionuclide therapy was successful.Conclusion:the expression of hTERT are different activity in glioma of U87and U251, therefore applying the hTERT as promoter guiding the specific expression of therapeutic genes in glioma cells carring the hNIS gene.buiding recombinant adenovirus, transferred glioma cells,enable them to make a strong iodine uptake ability, then mediate radioactive iodine killing tumor cells successfully.