Skip to main content
SpringerLink
Log in

Influence of age and parity on the susceptibility of rat mammary gland epithelial cells in primary cultures to 7,12-dimethylbenz(a) anthracene

  • Published:
In Vitro Aims and scope Submit manuscript

Summary

Mammary gland epithelial cells from rats of different ages or with different reproductive histories vary in their proliferative properties and susceptibility to dimethylbenz(a)anthracene (DMBA) carcinogenesis in vivo. The present study was carried out to determine whether these differences are maintained under in vitro conditions. Primary cultures of mammary gland epithelial cells of young virgin, old virgin, and parous rats were treated with various doses of DMBA. Growth rates, DNA synthesis, and dose-response curves were determined; the toxicity of DMBA was measured by its effect on cell growth. Cell morphology was studied by transmission and scanning electron microscopy. Epithelial cells from the mammary gland of young virgin rats adapted rapidly to the culture conditions, behaving as if the cells were in the logarithmic phase of growth prior to plating. Mammary gland epithelial cells from old virgin and parous rats required a lag period prior to cell growth during which the proliferating cells adapted to the culture conditions. Cells from each group had comparable doubling times, and DNA synthesis peaked approximately 1 d after initiation of growth in culture. The numbers of proliferating cells decreased with increasing age and parity of the donor. Mammary gland epithelial cells of young virgin rats were more susceptible to both low and high doses of DMBA than those of old virgin and parous rats when the carcinogen was added either 24 h after plating or at the peak of DNA synthesis. These results indicate that age and parity influence the proliferative status of the cells and their susceptibility to DMBA in vitro, simulating in that way the in vivo situation.

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

References

  1. Mazzaleni, G. F. Rapporti di frequenza fra carcinoma mammario e gravidanza. Riv. Anat. Oncol. 17:243–260; 1960.

    Google Scholar 

  2. Wynder, E. L.; Bross, I. J.; Hirayama, T. A study of the epidemiology of cancer of the breast. Cancer 13:559–601; 1960.

    Article  PubMed  CAS  Google Scholar 

  3. MacMahon, B. Etiology of human breast cancer. J. Natl. Cancer Inst. 50:21–42; 1973.

    PubMed  CAS  Google Scholar 

  4. Russo, J.; Saby, J.; Isenberg, W.; Russo, I. H. Pathogenesis of mammary carcinomas induced in rats by 7,12-dimethylbenz[a]anthracene. J. Natl. Cancer Inst. 59:435–445; 1977.

    Google Scholar 

  5. Huggins, C.; Briziarelli, G.; Sutton, H. Rapid induction of mammary carcinoma in the rat and the influence of hormones on the tumors. J. Exp. Med. 109:25–54; 1959.

    Article  PubMed  CAS  Google Scholar 

  6. Moon, R. C. Relationship between previous reproductive history and chemically induced mammary cancer in rats. Int. J. Cancer 4:312–317; 1969.

    Article  PubMed  CAS  Google Scholar 

  7. Russo, J.; Russo, I. H. DNA labeling index and structure of the rat mammary gland as determinants of its susceptibility to carcinogenesis. J. Natl. Cancer Inst. 61:1451–1459; 1978.

    PubMed  CAS  Google Scholar 

  8. Russo, J.; Russo, I. H. Influence of differentiation and cell kinetics on the susceptibility of the rat mammary gland to carcinogenesis. Cancer Res. 40:2677–2687; 1980.

    PubMed  CAS  Google Scholar 

  9. Cohen, L. A.; Tsuang, J.; Chang, P. C. Characterization of rat normal mammary epithelial cells and dimethylbenz[a]anthracene-induced mammary adenocarcinoma cells grown in monolayer culture. In Vitro 10:51–62; 1974.

    Article  PubMed  CAS  Google Scholar 

  10. Rivera, E. M.; Bern, H. A. Influence of insulin on maintenance and secretory stimulation of mouse mammary tissues by hormones in organ cultures. Endocrinology 69:340–353; 1961.

    Article  PubMed  CAS  Google Scholar 

  11. Mollenhauer, D. M. Plastic embedding mixtures for use in electron microscopy. Stain Technol. 39: 111–114; 1964.

    PubMed  CAS  Google Scholar 

  12. Lowry, O. H.; Rosebrough, N. J.; Farr, A. W.; Randall, R. J. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193: 265–275; 1951.

    PubMed  CAS  Google Scholar 

  13. Burton, K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem. M. 62:315–323; 1956.

    CAS  Google Scholar 

  14. Russo, J.; Furmanski, P.; Bradley, R.; Wells, P.; Rich, M. A. Differentiation of normal human mammary epithelial cells in culture: an ultrastructural study. Am. J. Anat. 145:57–78; 1976.

    Article  PubMed  CAS  Google Scholar 

  15. Russo, I. H.; Ireland, M.; Isenberg, W.; Russo, J. Ultrastructural description of three different epithelial cell types in rat mammary gland. Proc. Electron Microscopy Soc. Am. 34:146–147; 1976.

    Google Scholar 

  16. Hayflick, L. The limited in vitro lifetime of human diploid cell stains. Exp. Cell Res. 37:614–635; 1965.

    Article  PubMed  CAS  Google Scholar 

  17. Martin, G. M.; Sprague, C. A.; Epstein, C. J. Replicative life span of cultivated human cells. Effect of donor's age, tissue and genotype. Lab. Invest. 23:86–92; 1970.

    PubMed  CAS  Google Scholar 

  18. Goldstein, S.; Littlefield, J. N.; Saeldner, J. S. Diabetes mellitus and aging diminished plating efficiency of cultured human fibroblasts. Proc. Natl. Acad. Sci. USA 64:155–160; 1969.

    Article  PubMed  CAS  Google Scholar 

  19. Hayflick, L.; Moorhead, P. S. The serial cultivation of human diploid cell strains. Exp. Cell Res. 25:585–621; 1961.

    Article  Google Scholar 

  20. Schneider, E. L.; Mitsui, Y. The relationship between in vitro cellular aging and in vivo human age. Proc. Natl. Acad. Sci. USA 73:3584–3588; 1976.

    Article  PubMed  CAS  Google Scholar 

  21. Voyles, B. A.; McGrath, C. M. Differential response of malignant BALB/c mammary epithelial cells to the multiplication-stimulating activity of insulin. J. Natl. Cancer Inst. 62: 597–604; 1979.

    PubMed  CAS  Google Scholar 

  22. Bowman, P. D.; Meek, R. L.; Daniel, C. W. Aging of human fibroblasts in vitro; correlations between DNA synthetic ability and cell size. Exp. Cell Res. 93:184–190; 1975.

    Article  PubMed  CAS  Google Scholar 

  23. Tay, L. K.; Russo, J. Binding of 7,12-dimethylbenz[a]anthracene (DMBA) to DNA and subsequent induction of DNA excision repair in rat mammary epithelial cells in vitro. Proc. Am. Assoc. Cancer Res. 21:272a; 1980.

    Google Scholar 

  24. Russo, J.; Tay, L. K.; Wilgus, G. 7,12-Dimethylbenz[a]anthracene (DMBA) on rat mammary epithelial cells in culture. Fed. Proc. 38(3):1249; 1979.

    Google Scholar 

  25. Janss, D. H.; Moon, R. E.; Irving C. E. The binding of 7,12-dimethylbenz(a)anthracene to mammary parenchyma DNA and protein in vivo. Cancer Res. 32:254–258; 1972.

    PubMed  CAS  Google Scholar 

  26. Marquardt, H.; Sternberg S. S.; Phillips F. S. 7,12-Dimethylbenz(a)anthracene and hepatic neoplasia in regenerating rat liver. Chem. Biol. Interact. 2:401–403; 1970.

    Article  PubMed  CAS  Google Scholar 

  27. Prodi, G.; Rocchi, P.; Grilli, S. Binding of 7,12-dimethylbenz[a]anthracene and benzo(a)pyrene to nucleic acids and proteins of organs in rats. Cancer Res. 30:1020–1023; 1970.

    PubMed  CAS  Google Scholar 

  28. Tominaga, T.; Dao, T. L.; Libby, P. R. Effects of 7,12-dimethylbenz[a]anthracene on RNA polymerase in isolated mammary gland cell nuclei. Proc. Soc. Exp. Biol. Med. 136:694–697; 1971.

    PubMed  CAS  Google Scholar 

  29. Pound, A. W. Carcinogenesis and cell proliferation. New Zealand Med. J. 67:88–99; 1968.

    PubMed  Google Scholar 

  30. Shunkin, M. B.; Gruenstein, M.; Thatcher, D. Tritiated thymidine labeling of cells in rats following exposure to 7,12-dimethylbenz[a]anthracene. Cancer Res. 27:1494–1495; 1967.

    Google Scholar 

  31. Marquardt, H.; Bendick, A.; Philips, F. S.; Hoffman, D. Binding of [G-3H]7,12-dimethylbenz[a]anthracene to DNA of normal and of rapidly dividing hepatic cells of rats. Chem. Biol. Interact. 3:1–11; 1971.

    Article  PubMed  CAS  Google Scholar 

  32. Tominaga, T.; Libby, P. R.; Dao, T. L. An early effect of 7,12-dimethylbenz[a]anthracene on rat mammary gland DNA synthesis. Cancer Res. 30: 118–122; 1970.

    PubMed  CAS  Google Scholar 

  33. Tominaga, T.; Libby, P. R.; Dao, T. L. An early effect of 7,12-dimethylbenz[a]anthracene on rat mammary gland DNA synthesis. Cancer Res. 30: 118–122; 1970.

    PubMed  CAS  Google Scholar 

  34. Shunkin, M. B.; Gruenstein, M.; Thatcher, D. Tritiated thymidine labeling of cells in rats following exposure to 7,12-dimethylbenz[a]anthracene. Cancer Res. 27:1494–1495; 1967.

    Google Scholar 

  35. Russo, J.; Isenberg, W.; Ireland, M.; Russo, I. H. Ultrastructural changes in the mammary epithelial cell population during neoplastic development induced by a chemical carcinogen. Proc. Electron Microscopy Soc. Am. 34:250–251; 1976.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Experimental Pathology Laboratory, Department of Biology, Michigan Cancer Foundation, 110 E. Warren Avenue, 48201, Detroit, MI

    Jose Russo, Gerald Wilgus, Lawrence Tait & Irma H. Russo

Authors
  1. Jose Russo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gerald Wilgus
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lawrence Tait
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Irma H. Russo
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by Public Health Service Grants CA-23539 and CA-27026 from the National Cancer Institute and by an Institutional grant from the United Foundation of Greater Detroit.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Russo, J., Wilgus, G., Tait, L. et al. Influence of age and parity on the susceptibility of rat mammary gland epithelial cells in primary cultures to 7,12-dimethylbenz(a) anthracene. In Vitro 17, 877–884 (1981). https://doi.org/10.1007/BF02618283

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02618283

Key words

Search

Navigation