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Gene therapy of pancreatic cancer targeting the K-Ras oncogene

Cancer Gene Therapy volume 19, pages 862–869 (2012)Cite this article

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Abstract

Ras mutations are present in ∼95% of pancreatic cancer (PC) cases leading to increased proliferation and apoptosis resistance. The aim of this study is to selectively kill Ras-transformed cells by overexpressing the pro-apoptotic protein, p53 upregulated modulator of apoptosis (PUMA) under a Ras-responsive promoter. Colo357, Panc1 and MiaPaca, PC cell lines harboring K-Ras mutations, normal rat IEC18 enterocytes, and their K-Ras transformed R1 counterparts, were tested. We constructed adenoviral vectors containing the PUMA gene downstream to: (1) Four or five repetitive Ras-responsive elements (Ad-PY4/PY5-PUMA) and (2) a negative control (Ad-SV40-PUMA). Cell viability was estimated by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and apoptosis was evaluated by FACS. In vivo potency of the adenoviruses was evaluated in athymic nude mice. Infection with Ad-PY4/PY5-PUMA markedly inhibited cell growth (∼40–50%), and apoptosis was detected in all cells with high Ras activity, whereas IEC18 cells remained unaffected. The control vector, Ad-SV40-PUMA, did not induce any cell death. Selective and high expression of PUMA was detected in Ad-PY4-PUMA-infected cells. In vivo, Ad-PY4-PUMA inhibited by ∼35% the growth of established tumors compared with the Ad-SV40-PUMA. Selective overexpression of PUMA efficiently inhibits the growth of Ras-transformed cells while sparing the normal ones. This treatment modality may become a useful, effective and safe approach to selectively target Ras-mutated tumor cells.

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Acknowledgements

This study was partially supported by the Helen and Haim Lieber chair for cancer research and the Djerassi oncology center.

Author contributions

VL contributed to all the work described in this study was conducted by VL, writing of the manuscript, study concept and design. IN, SS, DK and AS contributed to technical support. NA and SK contributed to study supervision, study design, revision of the manuscript, professional counseling.

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Authors and Affiliations

  1. The Integrated Cancer Prevention Center, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel

    V Lisiansky, I Naumov, S Shapira, D Kazanov, N Arber & S Kraus

  2. Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel

    V Lisiansky, I Naumov, S Shapira, A Starr, N Arber & S Kraus

  3. Tel Aviv Sourasky Medical Center, Lung and Allergy Institute, Tel Aviv, Israel

    A Starr

  4. Department of Gastroenterology, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel

    N Arber

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  1. V Lisiansky
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Correspondence to N Arber.

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Lisiansky, V., Naumov, I., Shapira, S. et al. Gene therapy of pancreatic cancer targeting the K-Ras oncogene. Cancer Gene Ther 19, 862–869 (2012). https://doi.org/10.1038/cgt.2012.73

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  • DOI: https://doi.org/10.1038/cgt.2012.73

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