Abstract
Treatment of human gastric cancer clones in vitro with low doses of DFMO (5 mM) produced elongation of the cell population doubling times and lowering of the saturation densities. By contrast, DFMO treatment of normal human skin fibroblasts altered only the saturation density. The lack of an effect of 5 mM DFMO on the doubling time of normal fibroblasts may be directly related to baseline intracellular putrescine levels, which were about 2.5 times higher than in the cancer cells. The same dose of DFMO caused a rapid decrease in intracellular polyamine levels in the tumor clones. The effects on the doubling time and saturation density were almost totally abolished by the addition of 50 μM putrescine to the growth medium during the first 24 h of treatment with DFMO. Exposure to 5 mM DFMO for 24 h caused the human gastric cancer cells to become blocked in G1 phase only, and this led to a reduction in the fraction of cells in S phase. The G1 block was reversible and this cohort of cells eventually passed through S phase and then through G2 and M.
A higher 100 mM dose of DFMO and longer exposure times for both doses produced cell cycle changes and death of more than 90% of the cell population. These data suggest that cell kinetics changes observed under these experimental conditions may reflect polyamine-related alterations in the biochemical events of cell cycle progression kinetics; but may also be the result of DFMO-induced loss of cell viability.
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Abbreviations
- DFMO:
-
α-Difluoromethylornithine
- ODC:
-
ornithine decarboxylase
- ARA C:
-
cytosine arabinosidede
- MGBG:
-
methylglyoxal bis(guanylhydrazone)
References
Tobey RA, Gurley LR, Hildebrand CE, Ratliff RL, Walters RA: Sequential biochemical events in preparation for DNA replication and mitosis. In: Clarkson B, Baserga R (eds) Control of Proliferation in Animal Cells. Cold Spring Harbor, 1974, pp 665–679
Baserga R: The Cell Cycle and Cancer. Marcel Dekker, Inc., New York, 1971
Clarkson B, Baserga R: Control of proliferation in animal cells. Cold Spring Harbor, 1974
Mamont PS, Duchesne M-C, Grove J, Bey P: Antiproliferative properties of DL-α-difluoromethylornithine in cultured cells. A consequence of the irreversible inhibition of ornithine decarboxylase. Biochem Biophys Res Comm 81:58–66, 1978
Herbst EJ, Elliot QD: Role of polyamines in HeLa cell proliferation. Med Biol 59:410–416, 1981
Alhonen-hongisto L, Poso H, Janne J: Inhibition of polyamine accumulation and cell proliferation by derivatives of diaminopropane in Ehrlich ascites cells grown in culture. Biochem Biophys Acta 564:473–487, 1979
Heby O, Marton LJ, Wilson CB, Gray JW: Effect of methylglyoxal-Bis(Guanylhydrazone), an inhibitor of spermidine and spermine synthesis on cell cycle traverse. Euro J Cancer 13:1009–1017, 1977
Luk GD, Civin CI, Weissman RM, Baylin SB: Ornithine decarboxylase: essential in proliferation but not differentiation of human promyelocytic leukemia cells. Science 216:75–77, 1982
Porter CW, Bergeron RJ: Spermidine requirement for cell proliferation in eukaryotic cells: structural specificity and quantitation. Science 219:1083–1085, 1983
Prakash NJ, Schechter PJ, Mamont PS, Grove J, KochWeser J, Sjoerdsma A: Inhibition of EMT6 tumor growth by interference with polyamine biosynthesis; effects of α-difluoromethylornithine, an irreversible inhibitor of ornithine decarboxylase. Life Sciences 26:181–194, 1979
Gerner EW, Russel DH: The relationship between polyamine accumulation and DNA replication in synchronized Chinese hamster ovary cells after heat shock. Cancer Res 37:482–489, 1977
Sunkara PS, Prakash NJ, Chang CC, Sjoerdsma A: Cytotoxicity of methylglyoxal Bis(guanylhydrazone) in combination with α-difluoromethylornithine against HeLa cells and mouse L1210 leukemia. JNCI 70:505–509, 1983
Warrell Jr RP, Coonley CJ, Burchenal JH: Sequential inhibition of polyamine synthesis A phase I trial of DFMO (α-difluoromethylornithine) and methyl-GAG ‘methylglyoxal-bis(guanylhydrazone)’. Cancer Chemother Pharmacol 11:134–136, 1983
Barranco SC, Ford PJ, Townsend Jr CM: Heterogeneous survival responses of human gastric cancer clones to α-difluoromethylornithine in vitro. Invest New Drugs 4:337–345, 1986
Barranco SC, Townsend Jr CM, Casartelli C, Macik BG, Burger NL, Boerwinkle WR, Gourley WK: Establishment and characterization of an in vitro model system for human adenocarcinoma of the stomach. Cancer Res 43:1703–1709, 1983
Zante J, Schumann J, Barlogie B, Gohde W, Buchner T: New preparation and staining procedures for specific and rapid analysis of DNA distribution. In: Gohde W, Schumann J, Buchner T (eds) Pulse-Cytophotometry. European Press, Ghent, Belgium, pp 97–106, 1976
Guseman LF, Bryant J: Mathematic modeling and analysis of flow microfluorometry DNA distributions. In: D Lutz (ed) Pulse Cytophotometry, European Press, Ghent, Belgium, pp 79–92, 1978
Gerner EW, Stickney DG, Herman TS, Fuller DJM: Polyamines and polyamine biosynthesis in cells exposed to hyperthermia. Radiation Res 93:340–352, 1983
Prakash NJ, Sunkara PS: Combination chemotherapy involving α-difluoromethylornithine and 1-β-D-Arabinofuranosylcytosine in murine L1210 leukemia. Cancer Res 43:3192–3196, 1983
Sugiura M, Shafman T. Kufe D: Effects of polyamine depletion on proliferation and differentiation of murine erythroleukemia cells. Cancer Res 44:1440–1444, 1984
Gerner EW, Glass JR, Fuller DJM: Activation of ornithine decarboxylase in monolayer cells treated with trypsin. J Cell Physiol, 1985, in press
Heby O: Cell cycle phase specificity and therapeutic effectiveness of polyamine synthesis inhibitors. In: 13th International Cancer Congress, Part C: Biology of Cancer (2). Alan R Liss Inc, 150 Fifth Avenue, New York, pp 189–198, 1983
Mamont PS, Bey P, Koch-Weser J: Biochemical consequences of drug-induced polyamine deficiency in mammalian cells. In: Gaugas JM (ed) Polyamines in Biomedical Research, John Wiley and Sons, New York, 1980, p 147
Oredsson SM, Gray JW, Deen DF, Marton LJ: Decreased cytotoxicity of 1-β-D-Arabinofuranosylcytosine in 9L rat brain tumor cells pretreated with α-difluoromethylornithine in vitro. Cancer Res 43:2541–2544, 1983
Pohjanpelto P, Langstrom E, Linden M, Amehus S, Knuutila S, Holtta E, Heby O: Cell kinetics of a polyamine auxotrophic mammalian cell line as studied by flow cytometry. Proc Am Assoc Cancer Res 23:33–35, 1982
Seidenfeld J, Gray JW, Marton LJ: Depletion of 9L rat brain tumor cell polyamine content by treatment with DL-α-difluoromethylornithine inhibits proliferation and the G1 to S transition. Exp Cell Res 131:209, 1981
Sano Y, Deen DF, Oredsson SM, Marton LJ: Effects of α-difluoromethylornithine on the growth of 9L rat brain tumor multicellular spheroids and their response to 1,3-Bis-(2-chloroethyl)-1-nitrosourea. Cancer Res 144:577–581, 1984
Koch-Weser J, Schechter PJ, Bey P, Danzin C, Fozard JR, Jung MJ, Mamont PS, Seiler N, Prakash NJ, Sjoerdsma A: Potential of ornithine decarboxylase inhibitors as therapeutic agents. In: Morris DR, Marton LJ (eds) Polyamines in Biology and Medicine, Marcel Dekker, New York, 1981, p 437
Manni A, Wright C: Effect of Tamoxifen and α-difluoromethylornithine on clones of nitrosomethylureainduced rat mammary tumor cells grown in soft agar culture. Cancer Res 43:1084–1086, 1983
Cohen SS, Barner HD: Studies on unbalanced growth in Escherichia coli. Proc Natl Acad Sci U S 40:885–893, 1954
Abeloff MD, Slavik M, Luk GD, Griffin CA, Hermann J, Blanc O, Sjoerdsma A, Baylin SB: Phase 1 trial and pharmacokinetic studies of α-difluoromethylornithine — an inhibitor of polyamine biosynthesis. J Clin Oncol 2:124–130, 1984
Chang BK, Black Jr O, Gutman R: Inhibition of growth of human or hamster pancreatic cancer cell lines by -difluoromethylornithine alone and combined with cis-Diamminedichloroplatinum (II). Cancer Res 44:5100–5104, 1984
Siimes M, Seppanen P, Alhonen-hongisto L, Janne J: Synergistic action of two polyamine antimetabolites leads to a rapid therapeutic response in childhood leukemia. Int J Cancer 28:567–570, 1981
Sunkara PS, Prakash NJ, Rosenberger AL, Hagan AC, Lachmann PJ, Mayer GD: Potentiation of antitumor and antimetastatic activities of difluoromethylornithine by Interferon inducers. Cancer Res 44:2799–2802, 1984
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Barranco, S.C., Ford, P.J. & Townsend, C.M. Cell cycle kinetics responses of human stomach cancer cells to reduction in polyamine levels by treatment with αDifluoromethylornithine in vitro. Invest New Drugs 4, 347–357 (1986). https://doi.org/10.1007/BF00173507
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DOI: https://doi.org/10.1007/BF00173507