Skip to main content
SpringerLink
Log in

Toxicodynamics of tumour promoters of mouse skin

II. Bindung to protein kinase C of some new diterpene esters and induction of luminol-enhanced chemoluminescence in mouse peritoneal neutrophils

  • Original Papers
  • Experimental Oncology
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Summary

In binding competition assays using a protein kinase C preparation from mouse brain (particulate fraction)3H-labelled 12-O-tetradecanoylphorbol-13-acetate (TPA), for a series of new diterpene esters (DTE) the relative binding affinity [rba=K ai (TPA)/K ai (DTE)] in relation to TPA was determined. A wide range of values was noticed, some of the DTE binding more strongly than TPA (rba >1), others binding less strongly than TPA (rba <1) In comparative terms, competition for specific binding sites appears to correlate better with irritant than with promoting acitvity of the DTE. Using mouse peritoneal neutrophils, binding of [3H]-TPA was determined by a modification of the “cold-acetone filter assay”; saturation of high-affinity sites (K ad =0.2 nM) was obtained at concentrations ≦ 1 nM, but there was also evidence for specific binding at “low-affinity” sites (K ad =26 nM). Induction of chemoluminescence in the presence of luminol in mouse peritoneal neutrophils with a set of DTE usually elecited two peaks; at concentrations ≧10 nM DTE a short-lived, “spike-like” response lasting only from 0 to about 5 min (phase A) ist followed by a “plateau” response from about 5–120 min (phase B). This latter phase of chemoluminescence stimulation with luminol correlated well with theirritant potential of the DTE used. The sequence of the two phases can be inverted partially by using first TPA at 2,5 nM followed by a quick concentration increase to 100 nM; this indicates two different concentration-dependent events. As regards the intensity of the chemoluminescent response, quantitative but not qualitative differences between DTE were observed, which show some correlation with strong and weak tumour-promoting activity. Inhibition studies suggest the involvement of the myeloperoxidase/H2O2/Cl− system in the luminogenic response; it is suggested that the release of hypochlorite or a closely related oxidant may be instrumental in tumour promotion.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

DMSO:

dimethylsulphoxide

DTE:

diterpene ester

K10:

Gnidia factor K10 (Kraussianin)

K ai ,K ad :

apparent inhibition and dissociation constants

muPMN:

mouse peritoneal neutrophils

P2:

Pimelea factor P2

PDD:

phorbol-12,13-didecanoate

RPA:

12-O-retinoylphorbol-13-acetate

rba:

relative binding affinity

TPA:

12-O-tetradecanoylphorbol-13-acetate

3-TI:

3-O-tetradecanoylingenol

References

  • Allen RC (1981) Lucigenin chemiluminescence: a new approach to the study of polymorphonuclear leukocyte redox activity. In: Bioluminescence and chemoluminescence: basic chemistry and analytical applications. DeLuca and McElroy (eds) Academic Press, N.Y. pp 63–73

    Google Scholar 

  • Ashendel CL, Boutwell RK (1981) Direct measurement of specific binding of highly lipophilic phorbol diester to mouse epidermal membranes using cold acetone. Biochem Biophys Res Commun 99:543–549

    Google Scholar 

  • Bach H, Goerttler K (1971) Morphologische Untersuchungen zur hyperplasiogenen Wirkung des biologisch aktiven Phorbolesters A1. Virchows Arch [B] 8:196

    Google Scholar 

  • Blackshear PJ, Nemenoff RA, Hovis JG, Halsey DL, Stumpo DJ, Huang JK (1987) Insulin action in normal and protein kinase C-deficient rat hepatoma cells. Effects on protein phosphorylation, protein kinase activities, and ornithine decarboxylase activities and messenger ribonucleic acid leves. Mol Endocrinol 1:44–52

    Google Scholar 

  • Blumberg PM, Dunn JA, Jaken S, Jeng AY, Leach KL, Sharkey NA, Yeh E (1984) Specific receptors for phorbol ester tumour promoters and their involvement in biological responses. In: Slaga TJ (ed) Mechanisms of tumor promotion, vol III. CRC Boca Raton, Fla. pp 143–184

    Google Scholar 

  • Borek C, Troll W (1983) Modifiers of free radicals inhibit in vitro the oncogenic actions of x-rays, bleomycin, and the tumor promoter 12-O-tetradecanoylphorbol-13-acetate. Proc Natl Acad Sci USA 80:1304–1307

    Google Scholar 

  • Briheim G, Stendahl O, Dahlgren C (1984) Intra- and extracellular events in luminol-dependent chemiluminescence of polymor-phonuclear leukocytes. Infect Immun 45:1–5

    Google Scholar 

  • Cadenas E, Sies H (1984) Low-level chemoluminescence as an indicator of singlet molecular oxygen in biological systems. Methods Enzymol 105:221–231

    Google Scholar 

  • Cerutti PA (1985) Prooxidant states and tumor promotion. Science 227:375–381

    Google Scholar 

  • DeChatelet LR, Shirley PS, Johnston RB Jr (1976) Effect of phorbol myristate acetate on the oxidative metabolism of human polymorphonuclear leukocytes. Blood 47:545–554

    Google Scholar 

  • DeChatelet LR, Long GD, Shirley PS, Bass DA, Thomas MJ, Henderson FW, Cohen MS (1982) Mechanism of luminol-dependent chemoluminescence of human neutrophils. J Immunol 129:1589–1593

    Google Scholar 

  • Dutton DR, Bowden GT (1985) Indirect induction of a clastogenic effect in epidermal cells by a tumor promoter. Carcinogenesis 6:1279–1284

    Google Scholar 

  • Fellhauer M (1987) Dissertation, University of Heidelberg

  • Fellhauer M, Hecker E (1986) Screening of Thymelaeaceae species for irritant, cocarcinogenic and antineoplastic activity. Planta Med 5:73

    Google Scholar 

  • Frenkel K, Chrzan K (1987) Hydrogen peroxide formation and DNA base modification by tumor promoter-activated polymor-phonuclear leukocytes. Carcinogenesis 8:455–460

    Google Scholar 

  • Fürstenberger G, Berry DL, Sorg B, Marks F (1981) Skin tumor promotion by phorbol esters is a two-stage process. Proc Natl Acad Sci USA 78:7722–7726

    Google Scholar 

  • Fürstenberger G, Richter H, Argyris TS, Marks F (1982) Effects of the phorbol ester 4-O-methyl-12-O-tetradecanoylphorbol-13-acetate on mouse skin in vivo: evidence for its uselesness as a negative control compound in studies on the biological effects of phorbol ester tumor promoters. Cancer Res 42:342–348

    Google Scholar 

  • Fürstenberger G, Gross M, Marks F (1984) Involvement of prostaglandins in the process of skin tumor promotion. In: Thaler-Dao H et al. (eds) Icosanoids and cancer. Raven, N.Y., p 91

    Google Scholar 

  • Hafez A, Adolf W, Hecker E (1983) Active Principles of the Thymelaeaceae: III. Skin irritant and co-carcinogenic factors fromPimelea simplex. Planta Med 49:3–8

    Google Scholar 

  • Hecker E (1978) Structure-activity relationships in diterpene esters irritant and cocarcinogenic to mouse skin. In: Slaga TJ, Sivak A, Boutwell RK (eds) Carcinogenesis, vol 2. Mechanisms of tumor promotion and cocarcinogenesis. Raven, N.Y., pp 11–48

    Google Scholar 

  • Hecker E, Rippmann F (1988) General quantitative dose/time(response) relationship for solitary cancerogenesis and initiation/promotion in skin of NMRI-mice and its implications for risk assessment. Naunyn-Schmiedeberg's Arch Pharmacol 338:R11

    Google Scholar 

  • Hergenhahn M, Hecker E (1981) Specific binding of the tumor promoter TPA in various mouse organs as measured by a “cold acetone-filter assay”. Carcinogenesis 2:1277–1281

    Google Scholar 

  • Hergenhahn M, Fellhauer M, Kloz U, Tremp GL, Hecker E (1987) Release of reactive oxygen intermediates from mouse neutrophils by diterpene esters — TPA and RPA may differ in the induction of an unknown prostanoid. 4. SEK-Symposium Heidelberg, 18–21 March 1987, J Cancer Res Clin Oncol 113(Suppl):CANC 18, 16

    Google Scholar 

  • Keisari Y, Geva J, Flescher E, Goldin H, Lavie G (1985) Stimulation of human monocyte oxidative burst and related cytotoxicity by tumor-promoting and non-tumour-promoting diterpene esters, indole alkaloids and polyacetate type agents. Int J Cancer 36:467–472

    Google Scholar 

  • Kennedy AR, Troll W, Little JB (1984) Role of free radicals in the initiation and promotion of radiation transformation in vitro. Carcinogenesis 5:1213–1218

    Google Scholar 

  • Kensler TW, Trush MA (1981) Inhibition of phorbol ester-stimulated chemiluminescence in human polymorphonuclear leukocytes by retinoic acid and 5,6-epoxy-retinoic acid. Cancer Res 41:216–222

    Google Scholar 

  • Kensler TW, Trush MA (1984) Role of oxygen radicals in tumor promotion. Environ Mutagen 6:593–616

    Google Scholar 

  • Kikkawa U, Takai Y, Tanaka Y, Miyake R, Nishizuka Y (1983) Protein kinase C as a possible receptor protein of tumor promoting phorbol esters. J Biol Chem 258:11442–11445

    Google Scholar 

  • Kinzel V, Fürstenberger G, Loehrke H, Marks F (1986) Three-stage tumorigenesis in mouse skin: DNA synthesis as a prerequisite for the conversion induced by TPA prior to initiation. Carcinogenesis 7:779–782

    Google Scholar 

  • Kloz U, Hergenhahn M, Fellhauer M, Hecker E (1989) Induction of Epstein-Barr virus early antigens by tumor promoters of the diterpene ester type in Raji cells and specific (receptor) binding as compared to irritant and promoting activities. J Cancer Res Clin Oncol 115:148–156

    Google Scholar 

  • Lehrer RI, Cohen L (1981) Receptor-mediated regulation of superoxide production in human neutrophils stimulated by phorbol myrisate acetate. J Clin Invest 68:1314–1320

    Google Scholar 

  • Lewis JG, Adams DO (1985) Induction of 5,6-ring-saturated thymine bases in NIH-3T3 cells by phorbol ester-stimulated macro-phages: role of reactive oxygen intermediates. Cancer Res 45:1270–1275

    Google Scholar 

  • Lewis JG, Hamilton T, Adams DO (1986) The effect of macrophage development on the release of reactive oxygen intermediates and lipid oxidation products, and their ability to induce oxidative DNA damage in mammalian cells. Carcinogenesis 7:813–818

    Google Scholar 

  • Marnett LJ, Siedlik PH, Fung LWM (1982) Oxidation of phenidone and BW755 C by prostaglandin endoperoxide synthetase. J Biol Chem 257:6957–6964

    Google Scholar 

  • Marx JL (1987) Oxygen free radicals linked to many diseases. Science 235:529–531

    Google Scholar 

  • Nygaard OF, Simic MG (eds) (1983) Radioprotectors and anticarcinogens. Academic Press, N.Y.

    Google Scholar 

  • Ohkawa Y, Jwata K, Shibuya H, Fujiki H, Inui N (1984) A rapid, simple screening method for skin-tumor promoters using mouse peritoneal macrophages in vitro. Cancer Lett 21:253–260

    Google Scholar 

  • Ohkubo S, Yamada E, Endo T, Itoh H, Hidaka H (1984) Vitamin A acid-induced activation of Ca2+-activated, phospholipid-dependent protein kinase from rabbit retina. Biochem Biophys Res Commun 118:460–466

    Google Scholar 

  • Pontremoli S, Melloni E, Sparatore B, Michetti M, Salamino F, Horecker BL (1990) Isozymes of protein kinase C in human neutrophils and their modification by two endogenous proteinases. J Biol Chem 265:706–712

    Google Scholar 

  • Schmidt R, Hecker E (1989) Biological assays for irritant, tumor-initiating and tumour-promoting activities: II. Standardized initiation/promotion protocol and semiquantitative estimation of promoting (and initiating) potencies in skin of NMRI mice. J Cancer Res Clin Oncol 115:516–524

    Google Scholar 

  • Schwarz M, Peres G, Kunz W, Fürstenberger G, Kittstein W, Marks F (1984) On the role of superoxide anion radicals in skin tumour promotion. Carcinogenesis 5:1663–1670

    Google Scholar 

  • Sha'afi RI (1989) Some effects of phorbol esters are not mediated by protein kinase C. Biochem J 261:688

    Google Scholar 

  • Sorg B, Schmidt R, Hecker E (1987) Structure/activity relationships of polyfunctional diterpenes of the ingenane type: I. Tumour promoting activity of homologous, aliphatic 3-esters of ingenol and ofδ7,8-isoingenol-3-tetradecanoate. Carcinogenesis 8:1–4

    Google Scholar 

  • Taffet SM, Greenfield ARL, Haddox MK (1983) Retinal inhibits TPA activated, calcium-dependent, phospholipid-dependent protein kinase (“C” kinase). Biochem Biophys Res Commun 114:1194–1199

    Google Scholar 

  • Tremp GL (1986) Dissertation, University of Heidelberg

  • Tremp GL, Hecker E (1988) On the chemistry of phorbol: XIX. Preparation of tritium-labelled 12-O-retinoylphorbol-13-acetate [(20-3H]-RPA). Z Naturforsch 43:1672–1675

    Google Scholar 

  • Verma AK, Shapas BG, Rice HM, Boutwell RK (1979) Correlation of the inhibition by steroids of tumor promoter-induced mouse epidermal ornithine decarboxylase activity and of skin tumour promotion. Cancer Res 39:419–425

    Google Scholar 

  • Verma AK, Bryan GT, Reznikoff CA (1985) Tumor promotor 12-O-tetradecanoylphorbol-13-acetate receptors in normal human transitional epithelial cells. Carcinogenesis 6:427–432

    Google Scholar 

  • Wilson E, Olcott MC, Bell RM, Merrill AH, Lambeth JD (1986) Inhibition of the oxidative burst in human neutrophils by sphingoid long-chain bases. J Biol Chem 261:12616–12623

    Google Scholar 

  • Zayed S, Adolf W, Hecker E (1982) On the active principles of the Thymelaeaceae family: I. The irritants and cocarcinogens ofPimelea prostrata. Planta Med 45:67–77

    Google Scholar 

  • Zimmerman R, Cerutti P (1981) Active oxygen acts as a promoter of transformation in mouse embryo C3H/10T 1/2 /C18 fibroblasts. Proc Natl Acad Sci USA 81:2085–2087

    Google Scholar 

Download references

Author information

Author notes

  1. G. L. Tremp

    Present address: C/O Rhone-Poulenc Rorer S.A., Centre de Recherche de Vitry Alfortville, 13, Quai Jules Guesde, F-94403, Vitry sur Seine, France

Authors and Affiliations

  1. Institute of Biochemistry, Deutsches Krebs-Forschungszentrum, W-6900, Heidelberg, Federal Republic of Germany

    M. Hergenhahn, U. Kloz, M. Fellhauer, G. L. Tremp & E. Hecker

Authors
  1. M. Hergenhahn
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. U. Kloz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Fellhauer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. L. Tremp
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E. Hecker
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

See loc. cit. Kloz et al. 1989 for I. Comm. of this series

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hergenhahn, M., Kloz, U., Fellhauer, M. et al. Toxicodynamics of tumour promoters of mouse skin. J Cancer Res Clin Oncol 117, 385–395 (1991). https://doi.org/10.1007/BF01612756

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01612756

Key words

Search

Navigation