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Annals of Surgical Oncology

, Volume 5, Issue 8, pp 681–688 | Cite as

Adenovirus-mediated wild-typep53 tumor suppressor gene therapy induces apoptosis and suppresses growth of human pancreatic cancer

  • Michael Bouvet
  • Richard J. Bold
  • Julia Lee
  • Douglas B. Evans
  • James L. Abbruzzese
  • Paul J. Chiao
  • David J. McConkey
  • Joya Chandra
  • Sunil Chada
  • Bingliang Fang
  • Jack A. Roth
Original Articles

Abstract

Background: Thep53 tumor suppressor gene is mutated in up to 70% of pancreatic adenocarcinomas. We determined the effect of reintroduction of the wild-typep53 gene on proliferation and apoptosis in human pancreatic cancer cells using an adenoviral vector containing the wild-typep53 tumor suppressor gene.

Methods: Transduction efficiencies of six p53-mutant pancreatic cancer cell lines (AsPC-1, BxPC-3, Capan-1, CFPAC-1, MIA PaCa-2, and PANC-1) were determined using the reporter gene construct Ad5/CMV/β-gal. Cell proliferation was monitored using a3H-thymidine incorporation assay, Western blot analysis forp53 expression was performed, and DNA laddering and fluorescence-activated cell sorter analysis were used to assess apoptosis.p53 gene therapy was tested in vivo in a subcutaneous tumor model.

Results: The cell lines varied in transduction efficiency. The MIA PaCa-2 cells had the highest transduction efficiency, with 65% of pancreatic tumor cells staining positive for beta-galactosidase (β-gal) at a multiplicity of infection (MOI) of 50. At the same MOI, only 15% of the CFPAC-1 cells expressed the β-gal gene. Adenovirus-mediatedp53 gene transfer suppressed growth of all human pancreatic cancer cell lines in a dose-dependent manner. Western blot analysis confirmed the presence of the p53 protein product at 48 hours after infection. DNA ladders demonstrated increased chromatin degradation, and fluorescence-activated cell sorter analysis demonstrated a four-fold increase in apoptotic cells at 48 and 72 hours following infection with Ad5/CMV/p53 in the MIA PaCa-2 and PANC-1 cells. Suppression of tumor growth mediated by induction of apoptosis was observed in vivo in an established nude mouse subcutaneous tumor model following intratumoral injections of Ad5/CMV/p53.

Conclusions: Introduction of the wild-typep53 gene using an adenoviral vector in pancreatic cancer withp53 mutations induces apoptosis and inhibits cell growth. These data provide preliminary support for adenoviral mediatedp53 tumor suppressor gene therapy of human pancreatic cancer.

Key Words

p53 Pancreatic cancer Apoptosis Gene therapy 

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Copyright information

© The Society of Surgical Oncology, Inc. 1998

Authors and Affiliations

  • Michael Bouvet
    • 1
  • Richard J. Bold
    • 1
  • Julia Lee
    • 5
  • Douglas B. Evans
    • 1
  • James L. Abbruzzese
    • 2
  • Paul J. Chiao
    • 1
  • David J. McConkey
    • 3
  • Joya Chandra
    • 3
  • Sunil Chada
    • 5
  • Bingliang Fang
    • 4
  • Jack A. Roth
    • 4
  1. 1.From the Department of Surgical OncologyThe University of Texas M. D. Anderson Cancer CenterUSA
  2. 2.the Department of Medical OncologyThe University of Texas M. D. Anderson Cancer CenterUSA
  3. 3.the Department of Cell BiologyThe University of Texas M. D. Anderson Cancer CenterUSA
  4. 4.the Section of Thoracic Molecular Oncology, Department of Thoracic and Cardiovascular SurgeryThe University of Texas M. D. Anderson Cancer CenterUSA
  5. 5.Introgen TherapeuticsHouston

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