Retrovirus-mediated herpes simplex virus thymidine kinase gene transduction renders human thyroid carcinoma cell lines sensitive to ganciclovir and radiation in vitro and in vivo.
In an attempt to develop gene therapy for thyroid carcinomas, the present studies were undertaken to evaluate in vitro and in vivo therapeutic efficacy and toxicity of herpes simplex virus thymidine kinase (HSV-tk) gene and ganciclovir (GCV) treatment, a widely used prodrug/suicide gene therapy, in human thyroid carcinoma cell lines, FRO and WRO cells, using a means of retrovirus-mediated gene transduction. In vitro experiments demonstrated dose- and time-dependent cell killing by transduction of the HSV-tk gene followed by GCV treatment. The IC50 (the concentration required to elicit 50% growth inhibition) shifted from 250 to 0.5 mg/liter in FRO cells, and from 3,000 to 0.09 mg/liter in WRO cells with therapeutic indexes of 500 and 33,000, respectively. Treatment with 30 mg/liter GCV for 4 days led to complete cell death in HSV-tk tumor cells. Nontransduced cells mixed with transduced cells were also effectively killed by GCV (bystander effect). Low concentrations of GCV, which alone showed little cytotoxicity, enhanced radiation-induced cytotoxicity (radiosensitization). In vivo sc FRO-tk tumor models in nude mice also showed dose- and time-dependent tumor regression. The IC50 was less than 2 mg/kg, and treatment with 100 mg/kg GCV for 2 weeks completely eradicated all tumors. The bystander effect and radiosensitization were also obtained in vivo. These results suggest that the HSV-tk/GCV approach to human thyroid carcinoma cells appears to be very efficacious, with a wide therapeutic range, and exerts a bystander effect and radiosensitization both in vitro and in vivo. Thus, HSV-tk/GCV system, alone or in combination with radiotherapy, may be a promising suicide gene therapy for thyroid carcinomas.
Nishihara
E
E
Department of Nature Medicine, Atomic Bomb Disease Institute, Nagasaki University School of Medicine, Japan.
Nagayama
Y
Y
Mawatari
F
F
Tanaka
K
K
Namba
H
H
Niwa
M
M
Yamashita
S
S
D018263:Adenocarcinoma, Follicular / Q000473:pathology; D000818:Animals; D000998:Antiviral Agents / Q000627:therapeutic use*; D002277:Carcinoma / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D015774:Ganciclovir / Q000627:therapeutic use*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D011355:Prodrugs / Q000627:therapeutic use*; D011838:Radiation-Sensitizing Agents / Q000627:therapeutic use*; D012190:Retroviridae / Q000235:genetics; D018139:Simplexvirus / Q000201:enzymology*; D013937:Thymidine Kinase / Q000235:genetics*; D013964:Thyroid Neoplasms / Q000235:genetics / Q000532:radiotherapy* / Q000628:therapy*; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000528:radiation effects
D000998:Antiviral Agents; D011355:Prodrugs; D011838:Radiation-Sensitizing Agents; D013937:Thymidine Kinase; D015774:Ganciclovir