Procaspase 3 expression in ovarian carcinoma cells increases survivin transcription which can be countered with a dominant-negative mutant, survivin T34A; a combination gene therapy strategy.
Increased survivin expression is a negative prognostic marker in many tumours, including ovarian cancer. We show here that ovarian carcinoma cells upregulate survivin transcription in response to increased expression of the proapoptotic protein procaspase 3. We have utilized this observation in a combination gene therapy strategy using adenoviral constructs expressing the dominant-negative mutant survivin T34A (Ad Survivin T34A) and procaspase 3 (Ad Caspase 3) in ovarian carcinoma cell lines. Transfection of ovarian carcinoma cells with Ad Survivin T34A induces apoptosis via a caspase 9-mediated pathway that is not affected by cell cycle block prior to G2/M. Ad Survivin T34A-induced apoptosis can be significantly enhanced by cotransfection with Ad Caspase 3, and the combination of Ad Survivin T34A and Ad Caspase 3 leads to a significant increase in survival in a murine intraperitoneal ovarian carcinoma model with some long-term survivors. This suggests that inhibiting endogenous survivin activity while also delivering high levels of procaspase 3 allow proteolytic cleavage and activation of the terminal caspase cascade leading to tumour cell death.
McKay
Tristan R
TR
Cancer Research UK Molecular Oncology Unit, Imperial College School of Medicine, Hammersmith Hospital, London W12 0NN, UK.
Bell
Stephen
S
Tenev
Tencho
T
Stoll
Viola
V
Lopes
Rita
R
Lemoine
Nicholas R
NR
McNeish
Iain A
IA
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002277:Carcinoma / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle; D004354:Drug Screening Assays, Antitumor; D004792:Enzyme Precursors / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D005799:Genes, Dominant; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D051379:Mice; D008819:Mice, Nude; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D009154:Mutation; D009363:Neoplasm Proteins; D015739:Nocodazole / Q000494:pharmacology; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D015996:Survival Rate; D000077022:Survivin; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
C107489:BIRC5 protein, human; D004792:Enzyme Precursors; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D000077022:Survivin; C505407:CASP3 protein, human; C506433:CASP9 protein, human; C505409:Casp3 protein, mouse; C506434:Casp9 protein, mouse; D053148:Caspase 3; D053453:Caspase 9; D020169:Caspases; D015739:Nocodazole