Abstract
Bolesatine, a glycoprotein fromBoletus satanas Lenz, has previously been shown to be mitogenic in rat and human lymphocytes at very low concentrations, whereas higher concentrations inhibited protein synthesisin vitro and in severalin vivo systems. The low concentrations (1–10 ng/ml) of bolesatine were shown to activate protein kinase C (PKC)in vitro (cell-free system) and in Vero cells. In the same time, Vero cells significantly proliferated when incubated with bolesatine concentrations ranging from 1 to 10 ng/ml; the DNA synthesis increased by 27–59% as referred to the control, and InsP3 release increased in a concentration-dependent manner, up to 142%. At higher concentrations, 1–10 μg in cell-free systems, bolesatine inhibits protein synthesis by hydrolyzing the nucleoside triphosphates GTP and ATP.
In the present work, the implication of other toxic mechanisms, such as lipid peroxidation and active radical production, was investigated in relation to inhibition of cell growth, whereas possible modifications of the ratio m5dC/dC+m5dC were determined in order to correlate with the biphasic action of bolesatine in Vero cells.
Low concentrations of bolesatine up to 10 ng/ml do not increase malonaldehyde (MDA) production, while they induce hypomethylation (5.2% as compared to 7.1%). Higher concentrations (above 20 ng/ml) increase MDA production, from 58 ng/mg of cellular proteins to 113 ng/mg at a concentration of 50 ng/ml, for example, and induce hypermethylation in Vero cell DNA. It is concluded that low concentrations of bolesatine that are proliferative induce hypomethylation, which could be one of the pathways whereby bolesatine induces cell proliferation. Higher concentrations which enhance lipid peroxidation also induce hypermethylation. These mechanisms could be at least partly implicated in the pathway whereby bolesatine induces cell death.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- MDA:
-
malonaldehyde
- TBA:
-
thiobarbituric acid
References
Bedford MT, Van Helden PD. Hypermethylation of DNA in pathological conditions of human prostate. Cancer Res. 1987;47:5274–6
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.
Cedar H, Razin A. DNA methylation and development. Biochim Biophys Acta. 1990;1049:1–8
De Bustro A, Nelkin BD, Silverman A, Ehrlich G, Poiesz B, Baylin SB. The short arm of chromosome 11 is a ‘hot spot’ for hypermethylation in human neoplasia. Proc Natl Acad Sci USA. 1988;85:5693–7.
Eilers FI, Nelson IR. Characterization and partial purification of the toxin ofLepoita morganii. Toxicon. 1974;2:557–63.
Ennamany R, Kretz O, Badoc A, Deffieux G, Creppy EE. Effect of bolesatine, a glycoprotein fromBoletus satanas, on rat thymus in vivo. Toxicology. 1994;89:113–8.
Ennamany R, Lavergne JP, Reboud JP, Dirheimer G, Creppy EE. Mode of action of bolesatine, a toxic protein fromBoletus satanas Lenz. Mechanistic approaches. Toxicology. 1995a;100:51–5.
Ennamany R, Kretz O, Creppy EE. Effect of bolesatine on phospholipid/calcium dependent protein kinase in Vero cells and in rat thymus. Arch Toxicol. 1995b;69:624–30.
Jones PA. Gene activation by 5-azacytidine. In: Razin A, Cedar H, Riggs AD, eds. DNA methylation: biochemistry and biological significance. New York: Springer-Verlag; 1984: 165–88.
Jones PA, Wolkowicz MJ, Rideout WM, Gonzales FA, Marziasz CM, Tapscott SJ. De novo methylation of the myo D1 CpG island during the establishment of immortal cell lines. Proc Natl Acad Sci USA. 1990;87:6117–21.
Kretz O, Boulanger Y, Creppy EE, Dirheimer G. Purification and some properties of bolesatine, a protein inhibiting in vitro protein synthesis, from the mushroomBoletus satanas Lenz (Boletaceae). Arch Toxic Suppl. 1989;13:422–7.
Kretz O, Creppy EE, Dirheimer G. Characterization of bolesatine, a toxic protein from the mushroomBoletus satanas Lenz, and its effects on kidney cells. Toxicology. 1991a;66: 213–24.
Kretz O, Creppy EE, Dirheimer G. Disposition of the toxic protein, bolesatine, in rats: its resistance to proteolytic enzymes. Xenobiotica. 1991b;21:65–73.
Kretz O, Barbieri L, Creppy EE, Dirheimer G. Inhibition of protein synthesis in liver and kidney of mice by bolesatine: mechanistic approaches to the mode of action at molecular level. Toxicology. 1992;73:297–304.
Licastro F, Cristina M, Kretz O, Dirheimer G, Creppy EE, Stirp F. Mitogenic activity and immunological properties of bolesatine, a lectin isolated from the mushroomBoletus satanas Lenz. Int J Biochem. 1993;25:789–92.
Marshall CJ. Tumour suppressor genes. Cell. 1991;64:313–26.
Münzel PA, Pfohl-Leszkowicz A, Rohrdanz E, Keith G, Dirheimer G, Bock KW. Site-specific hypermethylation of c-myc protooncogene in liver nodules and inhibition of DNA methylation byN-nitrosomorpholine. Biochem Pharmacol. 1991;42:365–73.
Pfohl-Leszkowicz A, Dirheimer G. Changes in vivo DNA-(cytosine-5-)methyltransferase activity in oncogenetically susceptible rat target tissues induced byN-methyl-N-nitrosourea. Cancer Res. 1986;46:1110–3.
Razanamparany JL, Creppy EE, Perreau-Bertrand J, Boulanger Y, Dirheimer G. Purification and characterization of bolaffinine, a toxic protein fromBoletus affinis Peck (Boletaceae). Biochemistry. 1986;68:1217–23.
Schillings RT, Ruelius HW. Poricine, an acidic protein with antitumor activity from a basidiomycete. Arch Biochem Biophys. 1968;127:672–9
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ennamany, R., Marzetto, S., Saboureau, D. et al. Lipid peroxidation induced by bolesatine, a toxin ofBoletus satanas: Implication in m5dC variation in Vero cells related to inhibition of cell growth. Cell Biol Toxicol 11, 347–354 (1995). https://doi.org/10.1007/BF01305906
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01305906