AIM:To investigate the feasibility of radionuclide therapy of colon tumor cells by baculovirus vector-mediated transfer of the sodium/iodide symporter(NIS) gene.METHODS:A recombinant baculovirus plasmid carrying the N...AIM:To investigate the feasibility of radionuclide therapy of colon tumor cells by baculovirus vector-mediated transfer of the sodium/iodide symporter(NIS) gene.METHODS:A recombinant baculovirus plasmid carrying the NIS gene was constructed,and the viruses(BacNIS) were prepared using the Bac-to-Bac system.The infection efficiency in the colon cancer cell line SW1116 of a green fluorescent protein(GFP) expressing baculovirus(Bac-GFP) at different multiplicities of infection(MOI) with various concentrations of sodium butyrate was determined by flow cytometry.An in vitro cytotoxicity assay was also conducted after infection of SW1116 cells with Bac-NIS.Iodine uptake of Bac-NIS infected SW1116 cells and inhibition of this uptake by sodium perchlorate was examined,and the effect of Bac-NISmediated 131 I in killing tumor cells was evaluated by cell colony formation tests.RESULTS:Infection and transgene expression in SW1116with Bac-GFP were significantly enhanced by sodium butyrate,as up to 72% of SW1116 cells were infected with the virus at MOI of 400 and sodium butyrate at 0.5 mmol/L.No obvious cytotoxicity was observed under these conditions.Infection of SW1116 with Bac-NIS allowed uptake of 131 I in these tumor cells,which could be inhibited by sodium perchlorate.The viability of SW1116 cells infected with Bac-NIS was significantly lower than with Bac-GFP,suggesting that NIS gene-mediated 131 I uptake could specifically kill tumor cells.CONCLUSION:Baculovirus vector-mediated NIS gene therapy is a potential approach for treatment of colon cancer.展开更多
Radioiodine i s a routine therapy for differentiated thyroid cancers.Non-thyroid cancers can intake radioiodine after transfection of the human sodium iodide symporter (hNIS) gene.The human telomerase reverse transcri...Radioiodine i s a routine therapy for differentiated thyroid cancers.Non-thyroid cancers can intake radioiodine after transfection of the human sodium iodide symporter (hNIS) gene.The human telomerase reverse transcriptase (hTERT) promoter,an excellent tumor-specific promoter,has potential value for targeted gene therapy of glioma.We used the hTERT promoter to drive the expression of the hNIS and human thyroid peroxidase (hTPO) gene as a primary step for testing the effects of radioiodine therapy on malignant glioma.The U87 and U251 cells were co-transfected with two adenoviral vectors,in which the hNIS gene had been coupled to the hTERT promoter and the hTPO gene had been coupled to the CMV promoter,respectively.Then,we performed Western blot,125I intake and efflux assays,and clonogenic assay with cancer cells.We also did 99mTc tumor imaging of nude mice models.After co-transfection with Ad-hTERT-hNIS and Ad-CMV-hTPO,glioma cells showed the 125I intake almost 1.5 times higher than cells transfected with Ad-hTERT-hNIS alone.Western blots revealed bands of approximately 70 kDa and 110 kDa,consistent with the hNIS and hTPO proteins.In clonogenic assay,approximately 90% of cotransfected cells were killed,compared to 50% of control cells after incubated with 37 MBq of 131I.These results demonstrated that radioiodine therapy was effective in treating malignant glioma cell lines following induction of tumor-specific iodide intake by the hTERT promoter-directed hNIS expression in vitro.Cotransfected hNIS and hTPO genes can result in increased intake and longer retention of radioiodine.Nude mice harboring xenografts transfected with Ad-hTERT-NIS can take 99mTc scans.展开更多
基金Supported by Grants from the National Natural Science Foundation of China,No.30570525the Shanghai Leading Academic Discipline Project,No.S30203
文摘AIM:To investigate the feasibility of radionuclide therapy of colon tumor cells by baculovirus vector-mediated transfer of the sodium/iodide symporter(NIS) gene.METHODS:A recombinant baculovirus plasmid carrying the NIS gene was constructed,and the viruses(BacNIS) were prepared using the Bac-to-Bac system.The infection efficiency in the colon cancer cell line SW1116 of a green fluorescent protein(GFP) expressing baculovirus(Bac-GFP) at different multiplicities of infection(MOI) with various concentrations of sodium butyrate was determined by flow cytometry.An in vitro cytotoxicity assay was also conducted after infection of SW1116 cells with Bac-NIS.Iodine uptake of Bac-NIS infected SW1116 cells and inhibition of this uptake by sodium perchlorate was examined,and the effect of Bac-NISmediated 131 I in killing tumor cells was evaluated by cell colony formation tests.RESULTS:Infection and transgene expression in SW1116with Bac-GFP were significantly enhanced by sodium butyrate,as up to 72% of SW1116 cells were infected with the virus at MOI of 400 and sodium butyrate at 0.5 mmol/L.No obvious cytotoxicity was observed under these conditions.Infection of SW1116 with Bac-NIS allowed uptake of 131 I in these tumor cells,which could be inhibited by sodium perchlorate.The viability of SW1116 cells infected with Bac-NIS was significantly lower than with Bac-GFP,suggesting that NIS gene-mediated 131 I uptake could specifically kill tumor cells.CONCLUSION:Baculovirus vector-mediated NIS gene therapy is a potential approach for treatment of colon cancer.
基金supported by a grant from Tianjin Basic Research and Leading Edge Science Project of China (No. 08JCZDJC23900)
文摘Radioiodine i s a routine therapy for differentiated thyroid cancers.Non-thyroid cancers can intake radioiodine after transfection of the human sodium iodide symporter (hNIS) gene.The human telomerase reverse transcriptase (hTERT) promoter,an excellent tumor-specific promoter,has potential value for targeted gene therapy of glioma.We used the hTERT promoter to drive the expression of the hNIS and human thyroid peroxidase (hTPO) gene as a primary step for testing the effects of radioiodine therapy on malignant glioma.The U87 and U251 cells were co-transfected with two adenoviral vectors,in which the hNIS gene had been coupled to the hTERT promoter and the hTPO gene had been coupled to the CMV promoter,respectively.Then,we performed Western blot,125I intake and efflux assays,and clonogenic assay with cancer cells.We also did 99mTc tumor imaging of nude mice models.After co-transfection with Ad-hTERT-hNIS and Ad-CMV-hTPO,glioma cells showed the 125I intake almost 1.5 times higher than cells transfected with Ad-hTERT-hNIS alone.Western blots revealed bands of approximately 70 kDa and 110 kDa,consistent with the hNIS and hTPO proteins.In clonogenic assay,approximately 90% of cotransfected cells were killed,compared to 50% of control cells after incubated with 37 MBq of 131I.These results demonstrated that radioiodine therapy was effective in treating malignant glioma cell lines following induction of tumor-specific iodide intake by the hTERT promoter-directed hNIS expression in vitro.Cotransfected hNIS and hTPO genes can result in increased intake and longer retention of radioiodine.Nude mice harboring xenografts transfected with Ad-hTERT-NIS can take 99mTc scans.
