Journal of Neuro-Oncology

, Volume 19, Issue 1, pp 25–35 | Cite as

In vitro andin vivo cytotoxicity of gossypol against central nervous system tumor cell lines

  • Thomas Coyle
  • Sharon Levante
  • Michele Shetler
  • Jeffrey Winfield
Laboratory Investigation


Gossypol is a lipid soluble polyphenolic compound isolated from cotton seed oil which has been previously shown to have antiproliferative activityin vitro against a variety of human solid tumor cell lines. It has been extensively tested in clinical trials as a male contraceptive agent and found to be well tolerated. Its mechanism of action is thought to be inhibition of cellular energy metabolism. It inhibits glycolysis through inhibition of LDH isoenzyme type 5, and it inhibits mitochondrial oxidative phosphorylation and electron transport. We tested thein vitro antiproliferative effect of gossypol against four well characterized human glioma cell lines, HS 683, U373, U87 and U138, and one rat glioma cell line, C6, using the colorimetric Microculture Tetrazolium Assay (MTT). Cytotoxicity was found to be concentration and time dependent and increased with incubation times up to 8 days. The relative sensitivity of the glioma cell lines to gossypol at 48 hour incubation correlated with their respective LDH isoenzyme profiles, with the more sensitive cell lines expressing increased cathodal LDH isoenzymes (LDH 5). Thein vitro cytotoxicity of gossypol to these CNS tumor lines was compared to the other non central nervous system solid tumor cell lines which had been previously reported as being sensitive to gossypol, including SW-13 (adrenal), MCF-7 (breast), T47-D (breast), and HeLa (cervical). Additional lines tested included SK-MEL-3 (melanoma), Colo 201 (colon) and BRW, a line established in our laboratory from a patient with a Primitive Neuroectodermal tumor. C6, HS 683, and BRW had similar IC50s as the sensitive solid tumor cell lines. U373, U87 and U138 had significantly less sensitivity at 48 hours. There was greater cytotoxicity and no significant differences in the IC50s between any of cell lines at 8 day incubations. Additionally, we tested the cytotoxicity of gossypol against BRWin vivo, using the nude mouse xenograft model. Gossypol, given at a dose of 30 mg/kg per day five days a week for four weeks orally via gavage, was found to decrease the mean tumor weight of treated xenografts by more than 50% as compared to untreated xenografts. These findings suggest that gossypol has potential for further study as an agent for the treatment of primary CNS malignancies.

Key words

gossypol glioma human MTT assay lactate dehydrogenase isoenzymes xenograft 


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  1. 1.
    Black PM: Brain tumors. New Engl J Med 324: 1471–1476, 1991Google Scholar
  2. 2.
    Winger MJ, MacDonald DR, Cairncross JG: Supratentorial anaplastic gliomas in adults. The prognostic importance of extent of resection and prior low grade glioma. J Neurosurg 71: 487–493, 1985Google Scholar
  3. 3.
    Whittle IR, Denholm SW, Gregor A: Management of patients aged 60 years with supratentorial glioma: lessons from an audit. Surg Neurol 36: 106–111, 1991Google Scholar
  4. 4.
    Shapiro WR: Treatment of neuroectodermal brain tumors. Ann Neurol 12: 231–237, 1982Google Scholar
  5. 5.
    Rodriquez LA, Levin VA: Does chemotherapy benefit the patient with a central nervous system tumor? Oncology 1: 29–36, 1986Google Scholar
  6. 6.
    Rodriquez LA, Prados M, Silver P, Levin VA: Reevaluation of procarbazine for the treatment of recurrent malignant central nervous system tumors. Cancer 64: 2420–2429, 1989Google Scholar
  7. 7.
    Levin VA, Silver P, Hannigan J, Wara WM, Gutin PH, Davis RL, Wilson CB: Superiority of post radiotherapy adjuvant chemotherapy with CCNU, procarbazine, and vincristine (PCV) over BCNU for anaplastic gliomas: NCOG 6G61 final report. Int J Radiat Oncol Biol Phys 18: 321–324, 1990Google Scholar
  8. 8.
    Fine MA, Dear KGB, Loeffler JS, Bladi PM, Canellos GP: Metaanalysis of radiotherapy with and without adjuvant chemotherapy for malignant glioma in adults. Proc Amer Soc Clin Oncol 18: 125a, 1991Google Scholar
  9. 9.
    Rosenblum ML, Gerosa MA, Bodell WJ, Talcot RL: Tumor Cell Resistance. Prog Exp Tumor Res 27: 191–214, 1984Google Scholar
  10. 10.
    Maingiardi JR, Yodice P: Metabolism of the malignant astrocytoma. Neurosurg 26: 1–19, 1990Google Scholar
  11. 11.
    Sherwin AL, Le Blanc FE, McCann WP: Altered LDH isoenzymes in brain tumors. Arch Neurol 18: 311–315, 1968Google Scholar
  12. 12.
    Tuszynski G, Cossu G: Differential cytotoxic effect of gossypol in human melanoma, colon carcinoma, and other tissue culture cell lines. Cancer Res 44: 768–771, 1984Google Scholar
  13. 13.
    Benz C, Keniry M, Goldberg H: Selective toxicity of gossypol against epithelial tumors and its detection by magnetic resonance spectroscopy. Contraception 37: 221–228, 1988Google Scholar
  14. 14.
    Wu YW, Chik CL, Knazek RA: Anin vitro andin vivo study of antitumor effects of gossypol on human SW-13 adrenocortical carcinoma. Cancer Res 49: 3754–3758, 1989Google Scholar
  15. 15.
    Jaroszewski J, Kaplan O, Cohen J: Action of gossypol and rhodamine 123 on wild type and multidrug-resistance MCF-7 human breast cancer cells:31P nuclear magnetic resonance and toxicity studies. Cancer Res 50: 6936–6943, 1990Google Scholar
  16. 16.
    Tso WW, Lee CS: Gossypol uncoupling of respiratory chain and oxidative phosphorylation in ejaculated boar spermatozoa. Contraception 25: 649–655, 1982Google Scholar
  17. 17.
    Stephens DT, Critchlow LM, Hoskins DD: Mechanism of inhibition by gossypol of glycolysis and motility of monkey spermatozoain vitro. J Reprod Fert 69: 447–452, 1983Google Scholar
  18. 18.
    Lee C, Moon Y, Yuan J, Chen A: Enzyme inactivation and inhibition by gossypol. Mol Cell Biochem 47: 65–70, 1982Google Scholar
  19. 19.
    Qian S, Wang Z: Gossypol: a potential antifertility agent for males. Ann Rev of Pharm and Toxicol 24: 329–360, 1984Google Scholar
  20. 20.
    Stein RC, Joseph ACA, Matlin SA, Cunningham DC, Ford HT, Coombes RC: A Preliminary Clinical Study of gossypol in advanced human cancer. Cancer Chemother and Pharmacol 30: 480–482, 1992Google Scholar
  21. 21.
    Flack M, Pyle RG, Mullen NM, Lorenzo B, Wu YW, Knazek RA, Nisula BC, Reidenberg M: Oral gossypol in the treatment of metastatic adrenal cancer. J Clin Endocrinol Metabol 76: 1019–1024, 1993Google Scholar
  22. 22.
    Hansen MB, Nielsen SE, Berg K: Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Meth 119: 203–210, 1989Google Scholar
  23. 23.
    Jordan JP, Hand CM, Markowitz RS, Black P: Test for chemotherapeutic sensitivity of cerebral gliomas: use of colorimetric MTT assay. J of Neuro-Oncol 14: 19–35, 1992Google Scholar
  24. 24.
    Nikkhah G, Tonn JC, Hoffmann O, Kraemer HP, Darling JL, Schachenmayr W, Schonmayr R: The MTT assay for chemosensitivity testing for human tumors of the central nervous system. Part II: Evaluation of patient and drug-specific variables. J of Neuro-Oncol 13: 13–24, 1992Google Scholar
  25. 25.
    Houchens D, Ovejera A: ‘Experimental therapy in subcutaneous transplants’. In: Boven E, Winograd B (eds) The Nude Mouse in Oncology Research. CRC Press Boca Raton Fl, 1991, p 140Google Scholar
  26. 26.
    Benz C, Hollander C, Keniry M, James T, Mitchell M: Lactic dehydrogenase isoenzymes,31P magnetic resonance spectroscopy andin vitro antimitochondrial tumor toxicity with gossypol and rhodamine-123. J Clin Invest 79: 517–523, 1987Google Scholar
  27. 27.
    Pagliacci MC, Spinozzi F, Migliorati G, Fumi G, Smacchia M, Grignani F, Riccardi C, Nicoletti I: Genistein inhibits tumour cell growthin vitro, but enhances mitochondrial reduction of tetrazolium salts: a further pitfall in the MTT assay for evaluating cell growth and survival. Eur J Cancer 29A: 1573–1577, 1993Google Scholar
  28. 28.
    Benz C, Keniry M, Ford J, Townsend A, Cox F, Palayoor S, Matlin S, Hait W, Cowan K: Biochemical correlates of the antitumor and antimitochondrial properties of gossypol enantiomers. Mol Pharm 37: 840–847, 1990Google Scholar
  29. 29.
    Strom-Hasen T, Cornett C, Jaroszewski J: Interaction of gossypol with amino acids and peptides as a model of enzyme inhibition. Int J Peptide Protein Res 34: 306–310, 1989Google Scholar
  30. 30.
    Tso W: Gossypol inhibits ascites tumor cell proliferation. Cancer Lett 24: 257–261, 1984Google Scholar
  31. 31.
    Rao P, Wang Y, Lotzova E, Khan A, Rao S, Stephens LC: Antitumor effects of gossypol on murine tumors. Cancer Chemother and Pharmacol 15: 20–25, 1985Google Scholar
  32. 32.
    Coutinho E, Melo J: Clinical experience with gossypol in non-Chinese men: a follow up. Contraception 37: 137–151, 1988Google Scholar
  33. 33.
    Wu DF, Y YW, Tang ZM, Wang MZ: Pharmacokinetics of (+/-), (+) and (-) gossypol in humans and dogs. Clin Pharm Ther 39: 613–618, 1986Google Scholar
  34. 34.
    Carapella CM, Paggi MG, Cattani F, Ciottoli GB, Floridi A, Iandolo B, Raus AF: The potential role of lonidamine (LND) in the treatment of malignant glioma: phase II study. J Neuro-oncol 7: 103–108, 1989Google Scholar
  35. 35.
    Carapella C, Paggi M, Calvosa F, Cattani F, Jandolo B, Mastrostefano R, Raus L, Riccio A: Lonidamine in the combined treatment of malignant gliomas: A randomized study. J Neurosurg Sci 34: 261–264, 1990Google Scholar

Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Thomas Coyle
    • 1
    • 2
  • Sharon Levante
    • 2
  • Michele Shetler
    • 2
  • Jeffrey Winfield
    • 2
  1. 1.Department of Medicine of the SUNY Health Science Center at SyracuseUSA
  2. 2.Department of Neurosurgery of the SUNY Health Science Center at SyracuseUSA

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