L-Canavanine as a radiosensitization agent for human pancreatic cancer cells

  • Aimee K. Bence
  • Val R. Adams
  • Peter A. Crooks
Part of the Molecular and Cellular Biochemistry book series (DMCB, volume 40)


This study evaluated the in vitro effect of L-canavanine on cell cycle progression in the two human pancreatic cancer cells lines PANC-1 and MIA PaCa-2. After 72 h of exposure to L-canavanine, the percentage of cells in the radiosensitive G2/M phase of the cell cycle increased 6-fold in PANC-1 cells and 4-fold in MIA PaCa-2 cells, when compared to untreated cells. The capacity of L-canavanine to redistribute cells into the G2/M phase of the cell cycle was both concentration-and time-dependent. Since many drugs that cause cells to accumulate in the G2/M phase of the cell cycle are effective radiosensitization agents, the potential of L-canavanine to synergistically enhance the effects of ionizing radiation also was evaluated. The interaction between these treatment modalities was quantified using the median-effect equation and combination index analysis. LCanavanine was found to be synergistic with radiation when either PANC-1 or MIA PaCa-2 cells were exposed to L-canavanine for 72 h prior to irradiation. These results suggest that L-canavanine in combination with radiation may have clinical potential in the treatment of pancreatic cancer. (Mol Cell Biochem 244: 37-43, 2003)

Key words

L-canavanine cell cycle analysis ionizing radiation pancreatic cancer radiosensitization median-effect analysis 


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  1. 1.
    Greenlee RT, Hill-Harmon MB, Murray T, Thun M: Cancer statistics, 2001. CA Cancer J Clin 51: 15–39, 2001PubMedCrossRefGoogle Scholar
  2. 2.
    Warshaw AL, Fernandez-del Castillo C: Pancreatic carcinoma. N Engl J Med 326: 455–465, 1992PubMedCrossRefGoogle Scholar
  3. 3.
    Rosenthal GA: The biological effects and mode of action of L-canavanine, a structural analogue of L-arginine. Q Rev Biol 52: 155–178, 1977PubMedCrossRefGoogle Scholar
  4. 4.
    Thomas DA, Rosenthal GA, Gold DV, Dickey K: Growth inhibition of a rat colon tumor by L-canavanine. Cancer Res 46: 2898–2903, 1986PubMedGoogle Scholar
  5. 5.
    Green MH, Brooks TL, Mendelsohn J, Howell SB: Antitumor activity of L-canavanine against L1210 murine leukemia. Cancer Res 40: 535–537, 1980PubMedGoogle Scholar
  6. 6.
    Naha PM: An experimental model for selective inhibition of proliferating cells by chemotherapeutic agents. Cell Biol Int Rep 4: 829–839, 1980PubMedCrossRefGoogle Scholar
  7. 7.
    Naha PM, Silcock JM, Fellows L: Reappraisal of L-canavanine as an anti-tumour agent. Cell Biol Int Rep 4: 155–166, 1980PubMedCrossRefGoogle Scholar
  8. 8.
    Crooks PA, Rosenthal GA: Use of L-canavanine as a therapeutic agent for the treatment of pancreatic cancer. U.S. Patent 5,552,440. September 3, 1996Google Scholar
  9. 9.
    Thomas DA, Rosenthal GA: Toxicity and pharmacokinetics of the non-protein amino acid L-canavanine in the rat. Toxicol Appl Pharmacol 91: 395–405, 1987PubMedCrossRefGoogle Scholar
  10. 10.
    Rosenthal GA, Dahlman DL: L-Canavanine and protein synthesis in the tobacco hornworm Manduca sexta. Proc Natl Acad Sci USA 83: 14–18, 1986PubMedCrossRefGoogle Scholar
  11. 11.
    Rosenthal GA, Lambert J, Hoffmann D: Canavanine incorporation into the antibacterial proteins of the fly, Phormia terranovae (Diptera), and its effect on biological activity. J Biol Chem 264: 9768-9771, 1989PubMedGoogle Scholar
  12. 12.
    Rosenthal GA, Reichhart JM, Hoffmann JA: L-Canavanine incorporation into vitellogenin and macromolecular conformation. J Biol Chem 264: 13693–13696, 1989PubMedGoogle Scholar
  13. 13.
    Mattei E, Damasi D, Mileo AM, Delpino A, Ferrini U: Stress response, survival and enhancement of heat sensitivity in a human melanoma cell line treated with L-canavanine. Anticancer Res 12: 757–762, 1992PubMedGoogle Scholar
  14. 14.
    Laszlo A, Li GC: Effect of amino acid analogs on the development of thermotolerance and on thermotolerant cells. J Cell Physiol 154: 419–432, 1993PubMedCrossRefGoogle Scholar
  15. 15.
    Ackerman WW, Cox DC, Dinka S: Control of histones and DNA synthesis with canavanine, puromycin, and polio virus. Biochem Biophys Res Commun 19: 745–750, 1965CrossRefGoogle Scholar
  16. 16.
    Hare JD: Reversible inhibition of DNA synthesis by the arginine analog canavanine in hamster and mouse cells in vitro. Exptl Cell Res 58: 170–174, 1969PubMedCrossRefGoogle Scholar
  17. 17.
    Schactele CF, Rogers P: Canavanine death in Escherichia coll. J Mol Biol 34: 474–489, 1965CrossRefGoogle Scholar
  18. 18.
    Amorino GP, Freeman ML, Choy H: Enhancement of radiation effects in vitro by the estrogen metabolite 2-methoxyestradiol. Radiat Res 153: 384–391, 2000PubMedCrossRefGoogle Scholar
  19. 19.
    Buolamwini JK: Cell cycle molecular targets in novel anticancer drug discovery. Curr Pharm Des 6: 379–392, 2000PubMedCrossRefGoogle Scholar
  20. 20.
    Choy H, Rodriguez FF, Koester S, Hilsenbeck S, Von Hoff DD: Investigation of taxol as a potential radiation sensitizer. Cancer 71: 3774–3778, 1993PubMedCrossRefGoogle Scholar
  21. 21.
    Bodner WR, Hilaris BS, Mastoras DA: Radiation therapy in pancreatic cancer: Current practice and future trends. J Clin Gastroenterol 30: 230–233, 2000PubMedCrossRefGoogle Scholar
  22. 22.
    Green MH, Ward JF: Enhancement of human tumor cell killing by Lcanavanine in combination with y-radiation. Cancer Res 40: 4180–4182, 1983Google Scholar
  23. 23.
    Rosenthal GA: Preparation and colorimetric analysis of L-canavanine. Anal Biochem 77: 147–151, 1977PubMedCrossRefGoogle Scholar
  24. 24.
    Rosenthal GA: The preparation and colorimetric analysis of L-canaline. Anal Biochem 51: 354–361, 1973PubMedCrossRefGoogle Scholar
  25. 25.
    Bence AK, Crooks PA: Synthesis of L-indospicine. Syn Commun 32: 2075–2082, 2002CrossRefGoogle Scholar
  26. 26.
    Cory AH, Owen TC, Barltrop JA, Cory JG: Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Commun 3: 207–212, 1991PubMedGoogle Scholar
  27. 27.
    Chou TC: Derivation and properties of Michaelis-Menten type and Hill type equations for reference ligands. J Theor Biol 59: 253–276, 1976PubMedCrossRefGoogle Scholar
  28. 28.
    Chou TC, Talalay P: Quantitative analysis of dose-effect relationships: The combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22: 27–55, 1984PubMedCrossRefGoogle Scholar
  29. 29.
    Merlin JL: Concepts of synergism and antagonism. Anticancer Res 14: 2315–2319, 1994PubMedGoogle Scholar
  30. 30.
    Ding Y, Matsukawa Y, Ohtani Fujita N, Kato D, Dao S, Fujii T, Naito Y, Yoshikawa T, Sakai T, Rosenthal GA: Growth inhibition of A549 human lung adenocarcinoma cells by L-canavanine is associated with p21/WAF1 induction. Jpn J Cancer Res 90: 69–74, 1999PubMedCrossRefGoogle Scholar
  31. 31.
    Gu Y, Turck CW, Morgan DO: Inhibition of CDK2 activity in vivo by an associated 20K regulatory subunit. Nature 366: 707–710, 1993PubMedCrossRefGoogle Scholar
  32. 32.
    Redston MS, Caldas C, Seymour AB, Hruban RH, da Costa L, Yeo CJ, Kern SE: p53 mutations in pancreatic carcinoma and evidence of corn-mon involvement of homocopolymer tracts in DNA microdeletions. Cancer Res 54: 3025–3033, 1994PubMedGoogle Scholar
  33. 33.
    Bouvet M, Bold RJ, Lee J, Evans DB, Abbruzzese JL, Chiao PJ, McConkey DJ, Chandra J, Chada S, Fang B, Roth JA: Adenovirusmediated wild-type p53 tumor suppressor gene therapy induces apoptosis and suppresses growth of human pancreatic cancer. Ann Surg Oncol 5: 681–688, 1998PubMedCrossRefGoogle Scholar
  34. 34.
    Mogaki M, Hirota M, Chaney WG, Pour PM: Comparison of p53 protein expression and cellular localization in human and hamster pancreatic cancer cell lines. Carcinogenesis 14: 2589–2594, 1993PubMedCrossRefGoogle Scholar
  35. 35.
    Wang SR, Su HL, Chiu CC, Huang MH, Tsai CY, Tsai JJ, Yu CL: Alteration of intracellular DNA and RNA patterns by liver arginase studied with flow cytometry. Chung Hua I Hsueh Tsa Chih (Taipei) 50: 267–272, 1992Google Scholar
  36. 36.
    Schmidlin A, Wiesinger H: Transport of L-arginine in cultured glial cells. Glia 11: 262–268, 1994PubMedCrossRefGoogle Scholar
  37. 37.
    Rosenthal GA, Harper L: L-Homoarginine studies provide insight into the antimetabolic properties of L-canavanine. Insect Biochem Mol Biol 26: 389–394, 1996PubMedCrossRefGoogle Scholar
  38. 38.
    Swaffar DS, Ang CY: Growth inhibitory effect of L-canavanine against MIA PaCa-2 pancreatic cancer cells is not due to conversion to its toxic metabolite canaline. Anticancer Drugs 10: 113–118, 1999PubMedCrossRefGoogle Scholar
  39. 39.
    Terasima T, Tolmach LJ: X-ray sensitivity and DNA synthesis in synchronous populations of HeLa Cells. Science 140: 490–492, 1963PubMedCrossRefGoogle Scholar
  40. 40.
    Sinclair WK, Morton RA: X-ray sensitivity during the cell generation cycle of cultured Chinese hamster cells. Radiat Res 29: 450–474, 1966PubMedCrossRefGoogle Scholar
  41. 41.
    Brown KD, Lataxes TA, Shangary S, Mannino JL, Giardina JF, Chen J, Baskaran R: Ionizing radiation exposure results in up-regulation of Ku70 via a p53/ataxia-telangiectasia-mutated protein-dependent mechanism. J Biol Chem 275: 6651–6656, 2000PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Aimee K. Bence
    • 1
  • Val R. Adams
    • 2
  • Peter A. Crooks
    • 1
  1. 1.Division of Pharmaceutical SciencesUSA
  2. 2.Division of Pharmacy Practice and ScienceCollege of Pharmacy, University of KentuckyLexingtonUSA

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