Objective: To observe whether apoptosis could be induced by ganciclovir (GCV) in the herpes simplex virus-thymidine kinase gene-ganciclovir system (HSV-tk-GCV system) gene therapy for glioma.
Methods: Transduced C6/tk glioma cell as treated group, wild C6 cell as control group, were studied with morphology, gel-electrophoresis of DNA fragment analysis, fluoroscopy, and flow cytometric study before or after exposed to GCV, and apoptosis of transduced C6/tk glioma cell was ascertained.
Results: When C6/tk cell was treated with GCV at 5 micrograms/ml for 72 hours, cell apoptosis occurred. Typical apoptotic morphological features included cell shrinkage, condensation, and margination of nuclear chromatin were showed by light and electron microscopy, condensed nuclear chromatin and the fragment of nuclei were demonstrated by fluoroscopy, DNA ladder was showed by DNA fragment analysis, apoptotic peak was identified by flow cytometric study. The apoptotic cells accounted for 23% of the cell population. Apoptosis was not observed in control group treated by GCV.
Conclusions: The apoptosis of transduced C6/tk glioma was induced by GCV, which fit in with the basis of HSV-tk-GCV gene therapy for glioma.