Adenovirus-mediated thymidine kinase gene therapy induces apoptosis in human epithelial ovarian cancer cells and damages PARP-1.
Adenoviral (ADV) gene therapy with the thymidine kinase gene (TK) under control of the Rous sarcoma virus (RSV) promotor followed by the administration of acyclovir leads to replication errors in transcription and to cell death. This concept of ADV-RSV-TK has been established for the treatment of ovarian cancer cells. The purpose of this investigation was to clarify whether cell death after ADV-RSV-TK gene therapy and acyclovir administration is indeed due to apoptosis induction, whether the synergistic effect of ADV-RSV-TK gene therapy with chemotherapy was limited to the primary mechanism of action or whether the vector transduction itself exerted any pro-apoptotic effect was examined using the epithelial cell lines OVCAR-3 and MDAH-2774, established from human poorly differentiated serous ovarian cancer. Fluorimetric assay of caspase-3 activity was performed, as well as ELISA of the CK 18 split product M30. PARP cleavage was analysed by Western blotting. Apoptosis induction was established in this investigation as the mechanism of the ADV-RSV-TK gene therapy effect of acyclovir administration by caspase activity and subsequent CK 18 cleavage. Neither acyclovir nor vector administration alone showed any apoptotic activity. The synergistic effect of TK gene therapy and chemotherapeutic agents was shown to be TK induced. Significant anti-PARP 1 activity was found to be an ADV-RSV-TK treatment effect after acyclovir addition.
HELIOS Medical Center, Aue, Germany. dirk.kieback@helios-kliniken.de
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D000212:Acyclovir / Q000494:pharmacology; D000256:Adenoviridae / Q000235:genetics; D000998:Antiviral Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D045744:Cell Line, Tumor; D003131:Combined Modality Therapy; D018284:Cystadenocarcinoma, Serous / Q000235:genetics / Q000628:therapy*; D004357:Drug Synergism; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D010051:Ovarian Neoplasms / Q000235:genetics / Q000628:therapy*; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D052736:Rous sarcoma virus / Q000235:genetics; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics*
D000998:Antiviral Agents; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; D013937:Thymidine Kinase; D000212:Acyclovir