Summary
The biochemical effects of methyl chloride were investigated in tissues of F-344 rats and B6C3F1 mice (both sexes). Activities of GST were 2–3 times higher in livers of male B6C3F1 mice, compared with those of female mice, and with rats of both sexes. In kidneys GST activities of (male) mice were about 7 times lower than those found in livers. The activity of FDH was higher in livers of mice (both sexes) than in those of rats. No obvious sex difference was found in livers of rats and mice with respect to FDH. In kidneys, however, (minor) differences in FDH activities occurred between male and female B6C3F1 mice (4.7 vs. 3.1 nmol/min per mg). Sex differences of FDH activity in kidneys were not observed in F-344 rats. The microsomal transformation (by cytochrome P-450) of methyl chloride and S-methyl-L-cysteine to formaldehyde in tissues of B6C3F1 mice occurred preferentially in the liver. More formaldehyde was produced in liver microsomes of male, compared to those of female mice. Kidney microsomes metabolized methyl chloride to formaldehyde much less than liver microsomes. After a single exposure of mice of both sexes to 1000 ppm methyl chloride no elevation in formaldehyde concentrations was observed in livers and kidneys ex vivo. The determination of DNA lesions, using the alkaline elution technique, revealed no DNA-protein crosslinks in kidneys of male B6C3F1 mice after exposure to methyl chloride (1000 ppm, 6 h day-1, 4 days) and gave only minor evidence of singlestrand breaks. Lipid peroxidation (production of TBA reactive material), induced by single exposure to methyl chloride (1000 ppm, 6 h), was very pronounced in livers of male and female mice. Smaller increases in peroxidation were observed in the kidneys of exposed mice. The theory that renal tumors observed in male mice after chronic exposure of the test animals to high (1000 ppm) concentrations of methyl chloride, are evoked by intermediates and in situ produced formaldehyde is proven unlikely by our results.
Similar content being viewed by others
Abbreviations
- GST:
-
glutathione-S-transferase
- FDH:
-
formaldehyde dehydrogenase
- GSH:
-
glutathione
- CDNB:
-
1-chloro-2,4-dinitrobenzene (substrate for GST's)
- TBA:
-
Thiobarbituric acid (indicative of lipid peroxidation)
- TCA:
-
trichloroactetic acid
References
Andrews AW, Zawistowski ES, Valentine CR (1976) A comparison of the mutagenic properties of vinyl chloride and methyl chloride. Mutat Res 40:273–276
Anundi I, Högberg J, Stead AH (1979) Glutathione depletion in isolated hepatocytes: Its relation to lipid peroxidations and cell damage. Acta Pharmacol Toxicol 45:45–51
Battele Columbus Laboratories (1981) Final report on a chronic inhalation toxicology study in rats and mice exposed to methyl chloride submitted to the Chemical Industry Institute of Toxicology, vol 1–4. See also: Toxicologist 2:161 (1982) (abstract)
Bus JS (1982) Integrated studies of methyl chloride toxicity. CIIT Activities 2(1):3–4
Chausseaud LF (1979) The role of glutathione-S-transferase in metabolism of chemical carcinogens and other electrophilic agents. In: Klein G, Weinhouse S (eds) Advances in cancer research vol 29. Academic Press, New York, pp 175
DFG, Deutsche Forschungsgemeinschaft (1984) Methylchlorid/Chlormethan. In: Henschler D (ed) Toxikologisch-arbeitsmedizinische Begrüdungen von MAK-Werten. Verlag Chemie, Weinheim
Fornace Jr AJ, Little JB (1979) DNA-protein crosslinking by chemical carcinogens in mammalian cells. Cancer Res 39:704–710
Grafstrom RC, Fornace Jr AJ, Autrup H, Lechner JF, Harris CC (1983) Formaldehyde damage to DNA and inhibition of DNA repair in human bronchial cells. Science 220:216–218
Habig WH, Pabst MJ, Jacoby WB (1974) Glutathione-S-transferases — the first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139
Heck HDA, White EL, Casanova-Schmitz M (1982) Determination of formaldehyde in biological tissues by gas chromatography/mass spectrometry. Biomed Mass Spectrom 9:347–353
Huber W (1981) Die Mechanisierung der photometrischen Analyse mit den Bausteinen der Flüssigkeits-Chromatographie. 2. Mitteilung. Schnelle Präzisionsbestimmung für Formaldehyd. Fresenius Z Anal Chem 309:386–390
Kitabchi AE, Challoner DR, Williams RH (1968) Respiration and lipid peroxidation in tocopherol deficient rat hearts. Proc Soc Exp Med Biol 127:647–650
Kohn KW (1979) DNA as a target in cancer chemotherapy: measurement of macromolecular DNA damage produced in mammalian cells by anticancer agents and carcinogens. Methods Cancer Res vol XVI:291–340
Kohn KW, Erickson LC, Ewig RAG, Friedman CA (1976) Fractionation of DNA from mammalian cells by alkaline elution. Biochemistry 15:4629–4637
Kornbrust DJ, Bus JS (1982) Metabolism of methyl chloride to formate in rats. Toxicol Appl Pharmacol 65:135–143
Kornbrust DJ, Bus JS (1983) The role of glutathione and cytochrome P-450 in the metabolism of methyl chloride. Toxicol Appl Pharmacol 67:246–256
Kornbrust DJ, Bus JS (1984) Glutathione depletion by methyl chloride and association with lipid peroxidation in mice and rats. Toxicol Appl Pharmacol 72:388–399
Kornbrust DJ, Bus JS, Doerjer G, Swenberg JA (1982) Association of inhaled 14C-methyl chloride with macromolecules from various rat tissues. Toxicol Appl Pharmacol 72:388–399
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Nash T (1953) The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochem J 55:416–421
Peter H, Laib RJ, Ottenwälder H, Topp H, Rupprich N, Bolt HM (1985) DNA-binding assay of methyl chloride. Arch Toxicol 57:84–87
Remmer H, Greim H, Schenkman JB, Estabrook RW (1967) Methods for the elevation of hepatic microsomal mixed function oxidase levels and cytochrome P-450. Methods Enzymol 10:703–708
Simmon VF, Kanhanen K, Mortelmans K, Tardiff R (1977) Mutagenic activities of chemicals identified in drinking water. In: Scott D, Bridges BA, Sobels FA (eds) Progress in genetic toxicology. Elsevier/North Holland Biomedical Press, Amsterdam, pp 249–258
Sina JF, Bean LC, Dysart GR, Taylor VI, Bradley MO (1983) Evaluation of the alkaline elution/rat hepatocyte assay as a predictor of carcinogenic/mutagenic potential. Mutat Res 113:357–391
Sterzel W, Bedford P, Eisenbrand G (1985) Automated determination of DNA using the fluorochrome Hoechst 33258. Anal Biochem 147:462–467
Uotila L, Koivusalo M (1981) Formaldehyde dehydrogenase. In: Jakoby WB (ed) Methods in enzymology vol 77: detoxication and drug metabolism: conjugation and related systems. Academic Press, New York, pp 314–320
Younes M, Siegers C-P (1980) Lipid peroxidation as a consequence of glutathione depletion in rat and mouse liver. Res Commun Chem Pathol Pharmacol 27:119–127
Author information
Authors and Affiliations
Additional information
Dedicated to Professor Werner Kunz on occasion of his 65th birthday
Rights and permissions
About this article
Cite this article
Jäger, R., Peter, H., Sterzel, W. et al. Biochemical effect of methyl chloride in relation to its tumorigenicity. J Cancer Res Clin Oncol 114, 64–70 (1988). https://doi.org/10.1007/BF00390487
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00390487