Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Methioninase gene therapy with selenomethionine induces apoptosis in bcl-2-overproducing lung cancer cells

Cancer Gene Therapy volume 10, pages 445–450 (2003)Cite this article

Abstract

We have previously shown that the toxic pro-oxidant methylselenol is released from selenomethionine (SeMET) by cancer cells transformed with the adenoviral methionine α,γ-lyase (methioninase, MET) gene cloned from Pseudomonas putida. Methylselenol damaged the mitochondria via oxidative stress, and caused cytochrome c release into the cytosol thereby activating caspase enzymes and thereby apoptosis. However, gene therapy strategies are less effective if tumor cells overexpress the antiapoptotic mitochondrial protein bcl-2. In this study, we investigated whether rAdMET/SeMET was effective against bcl-2-overproducing A549 lung cancer cells. We established two clones of the human lung cancer A549 cell line that show moderate and high expression levels of bcl-2, respectively, compared to the parent cell line, which has very low bcl-2 expression. Staurosporine-induced apoptosis was inhibited in the bcl-2-overproducing clones as well as in the parental cell line. In contrast to staurosporine, apoptosis was induced in the bcl-2-overproducing clones as well as the parental cell line by AdMET/SeMET. Apoptosis in the rAdMET–SeMET-treated cells was determined by fragmentation of nuclei, and release of cytochrome c from mitochondria to the cytosol. A strong bystander effect of AdMET/SeMET was observed on A549 cells as well as the bcl-2-overproducing clones. rAdMET/SeMET prodrug gene therapy is therefore a promising novel strategy effective against bcl-2 overexpression, which has blocked other gene therapy strategies.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  1. Miki K, Xu M, Gupta A, et al. Methioninase cancer gene therapy with selenomethionine as suicide prodrug substrate. Cancer Res. 2001;61:6805–6810.

    CAS  PubMed  Google Scholar 

  2. Wolter KG, Hsu YT, Smith CL, Nechushtan A, Xi XG, Youle RJ . Movement of Bax from the cytosol to mitochondria during apoptosis. J Cell Biol. 1997;139:1281–1292.

    Article  CAS  Google Scholar 

  3. Kroemer G, Reed JC . Mitochondrial control of cell death. Nat Med. 2000;6:513–519.

    Article  CAS  Google Scholar 

  4. Costantini P, Jacotot E, Decaudin D, Kroemer G . Mitochondrion as a novel target of anticancer chemotherapy. J Nat Cancer Inst. 2000;92:1042–1053.

    Article  CAS  Google Scholar 

  5. Susin SA, Zamzami N, Castedo M, et al. Bcl-2 inhibits the mitochondrial release of an apoptogenic protease. J Exp Med. 1996;184:1331–1341.

    Article  CAS  Google Scholar 

  6. Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD . The release of cytochrome c from mitochondria: a primary site for Bcl-2 reguration of apoptosis. Science. 1997;275:1132–1136.

    Article  CAS  Google Scholar 

  7. Yang J, Liu X, Bhalla K, et al. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science. 1997;275:1129–1132.

    Article  CAS  Google Scholar 

  8. Weller M, Malipiero U, Aguzzi A, Reed JC, Fontana A . Protooncogene bcl-2 gene transfer abrogates Fas/APO-1 antobody-mediated apoptosis of human malignant glioma cells and confers to resistance to chemotherapeutic drugs and therapeutic irradiation. J Clin Invest. 1995;95:2633–2643.

    Article  CAS  Google Scholar 

  9. Hamel W, Magnelli L, Chiarugi VP, Israel MA . Herpes simplex virus thymidine kinase/ganciclovir-mediated apoptosis death of bystander cells. Cancer Res. 1996;56:2697–2702.

    CAS  PubMed  Google Scholar 

  10. Carsten F, Christoph S, Barbara H, et al. Bcl-2 expression in high-grade human glioma: clinical and experimental study. J Neurooncol. 2000;48:207–216.

    Article  Google Scholar 

  11. Tu Y, Xu FH, Liu J, et al. Upregulated expression of BCL-2 in multiple myeloma cells induced by exposure to doxorubicin, etoposide, and hydrogen peroxide. Blood. 1996;88:1805–1812.

    CAS  PubMed  Google Scholar 

  12. Elstein KH, Zucker RM . Comparison of cellular and nuclear flow cytometirc techniques for discriminating apoptotic subpopulations. Exp Cell Res. 1994;211:322–331.

    Article  CAS  Google Scholar 

  13. Susin SA, Lorenzo HK, Zamzami N, et al. Mitochondrial release of caspase-2 and -9 during the apoptotic process. J Exp Med. 1999;189:381–394.

    Article  CAS  Google Scholar 

  14. Susin SA, Lorenzo HK, Zamzami N, et al. Molecular characterization of mitochondrial apoptosis-inducing factor. Nature. 1999;397:441–446.

    Article  CAS  Google Scholar 

  15. Nutt LK, Chandra J, Pataer A, et al. Bax-mediated Ca2+ mobilization promotes cytochrome c release during apoptosis. J. Biol Chem. 2002;277:20301–20308.

    Article  CAS  Google Scholar 

  16. Spallholz JE . On the nature of selenium toxicity and carcinostatic activity. Free Radic Biol Med. 1994;17:45–64.

    Article  CAS  Google Scholar 

  17. Stewart MS, Spallholz JE, Nelder KH, Pence BC . Selenium compounds have disparate abilities to impose oxidative stress and induce apoptosis. Free Radic Biol Med. 1999;26:42–48.

    Article  CAS  Google Scholar 

  18. Clark LC, Combs GF, Turnbull BW, et al. Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin. A randomized controlled trial. JAMA. 1996;276:1957–1963.

    Article  CAS  Google Scholar 

  19. Chaudiere J, Courtin O, Leclaire J . Glutathione oxidase activity of selenocystamine: a mechanistic study. Arch Biochem Biophys. 1992;296:328–336.

    Article  CAS  Google Scholar 

  20. Yan L, Spallholz E . Generation of reactive oxygen species from the reaction of selenium compounds with thiols and mammary tumor cells. Biochem Pharmacol. 1993;45: 429–437.

    CAS  PubMed  Google Scholar 

  21. Green DR, Reed JC . Mitochondria and apoptosis. Science. 1998;281:1309–1312.

    Article  CAS  Google Scholar 

  22. Cai J, Yang J, Jones DP . Mitochondrial control of apoptosis: The role of cytochrome c. Biochim Biophys Acta. 1998;1366:139–149.

    Article  CAS  Google Scholar 

  23. Li P, Nijhawan D, Budihardjo I, et al. Cytochrome c and dATP-dependent formation of Apaf–caspase-9 complex initiates an apoptotic protease cascades. Cell. 1997;91:479–489.

    Article  CAS  Google Scholar 

  24. Thornberry NA, Lazebnik Y . Caspase: enemies within. Science. 1998;281:1312–1316.

    Article  CAS  Google Scholar 

  25. Tanaka H, Esaki N, Soda K . Properties of L-methionine γ-lyase from Pseudomonas ovalis. Biochemistry. 1977;16:100–106.

    Article  CAS  Google Scholar 

  26. Inoue H, Inagaki K, Sugimoto M, Esaki N, Soda K, Tanaka H . Structure analysis of the L-methionine γ-lyase gene from Pseudomonas putida. J Biochem. 1995;117:1120–1125.

    Article  CAS  Google Scholar 

  27. Esaki N, Tanaka H, Uemura S, Suzuki T, Soda K . Catalytic action of L-methionine gamma-lyase on selenomethionine and selenols. Biochemistry. 1979;18:407–410.

    Article  CAS  Google Scholar 

  28. Akhaven-Tafti H, et al. Clin Chem. 1995; 41:1368–1369.

  29. Iwahashi H, Eguchi Y, Yasuhara N, et al. Synergistic anti-apoptotic activity between Bcl-2 and SMN implicated in spinal atrophy. Nature. 1997;390:413–417.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. AntiCancer, Inc., 7917 Ostrow Street, San Diego, 92111, California, USA

    Norio Yamamoto, Anshu Gupta, Mingxu Xu, Kenji Miki & R M Hoffman

  2. Department of Surgery, University of California at San Diego, 200 West Arbor Drive, San Diego, 92103-8220, California, USA

    Norio Yamamoto, Anshu Gupta, A R Moossa & R M Hoffman

  3. Department of Orthopedic Surgery, Graduate School of Medicine, Kanazawa University, 13-1 Takaramachi, Kanazawa, 920-8641, Japan

    Norio Yamamoto, Hiroyuki Tsuchiya & Katsuro Tomita

  4. Department of Surgery, Graduate School of Medicine, University of Tokyo, Japan

    Kenji Miki

  5. Graduate School of Medicine, Osaka University, Japan

    Yoshihide Tsujimoto

Authors
  1. Norio Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anshu Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mingxu Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kenji Miki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yoshihide Tsujimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hiroyuki Tsuchiya
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Katsuro Tomita
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. A R Moossa
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. R M Hoffman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to R M Hoffman.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yamamoto, N., Gupta, A., Xu, M. et al. Methioninase gene therapy with selenomethionine induces apoptosis in bcl-2-overproducing lung cancer cells. Cancer Gene Ther 10, 445–450 (2003). https://doi.org/10.1038/sj.cgt.7700587

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.cgt.7700587

Keywords

This article is cited by

Search

Advanced search

Quick links