Skip to main content
SpringerLink
Log in

Proliferation and differentiation of prepubertal mouse vaginal epithelial cells in vitro and the specificity of estrogen-induced growth retardation

  • Regular Papers
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

Mouse vaginal epithelial cells were isolated from intact 21-day-old BALB/cCrgl mice and cultured in a serum-free medium (SF20: basal medium supplemented with insulin, epidermal growth factor, transferrin, and bovine serum albumin—fraction V) to examine the proliferation, differentiation, and specificity of estrogen-induced growth retardation in vitro. Histologic and ultrastructural studies showed that vaginal epithelial cells undergo differentiative changes in vitro in the absence of estrogen, and that these changes are similar to those induced in vivo by estrogen. Addition of 17β-estradiol inhibited cellular proliferation in a dose-dependent manner. Whereas other estrane derivatives (17α-estradiol and estriol) also significantly retarded cellular proliferation, cholesterol, testosterone, and progesterone had no effect. Keoxifene, an antiestrogen, significantly reversed estrogen-induced growth inhibition, resulting in proliferation of estrogen-treated cells equivalent to that of the untreated control. The results suggest that both proliferation and differentiation of prepubertal mouse vaginal epithelial cells in vitro are estrogen-independent, and that the growth inhibition is a specific estrogen-induced response.

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. Adler, A. J.; Nelson, J. F. Aging and chronic exposure impair estradiol-induced cornification but not proliferation of vaginal epithelium in C57BL/6J mice. Biol. Reprod. 38:175–182; 1988.

    Article  PubMed  CAS  Google Scholar 

  2. Bern, H. A. Is estrogen a cellular signal for female genital tract epithelium? In: Lavia, L. A., ed. Cellular signals controlling uterine function. New York: Plenum. In press; 1991.

    Google Scholar 

  3. Bindal, R. D.; Carlson, K. E.; Katzenellenbogen, B. S., et al. Lipophilic impurities, not phenolsulfonphthalein, account for the estrogenic activity in commercial preparations of phenol red. J. Steroid Biochem. 31:287–293; 1988.

    Article  PubMed  CAS  Google Scholar 

  4. Black, L. J.; Jones, C. D.; Falcone, J. F. Antagonism of estrogen action with a new benzothiophene derived antiestrogen. Life Sci. 32:1031–1036; 1982.

    Article  Google Scholar 

  5. Buckley, A.; Davidson, J. M.; Kamerath, C. D., et al. Sustained release of epidermal growth factor accelerates wound repair. Proc. Natl. Acad. Sci. USA 82:7340–7344; 1985.

    Article  PubMed  CAS  Google Scholar 

  6. Conti, C. J.; Tasat, D. R. Regulation of cultured rat vaginal epithelial cells by 17β-estradiol and progesterone. J. Steroid Biochem. 24:747–751; 1986.

    Article  PubMed  CAS  Google Scholar 

  7. Cooke, P. S.; Uchima, F.-D. A.; Fujii, D. K., et al. Restoration of normal morphology and estrogen responsiveness in cultured vaginal and uterine epithelia transplanted with stroma. Proc. Natl. Acad. Sci. USA 83:2109–2113; 1986.

    Article  PubMed  CAS  Google Scholar 

  8. Cunha, G. R.; Fujii, H. Stromal-parenchymal interactions in normal and abnormal development of the genital tract. In: Herbst, A. L.; Bern, H. A., eds. Developmental effects of diethylstilbestrol (DES) in pregnancy. New York: Thieme-Stratton; 1981:179–193.

    Google Scholar 

  9. Eiger, S.; Mills, K. T.; Bern, H. A. Steroid binding alterations in tissue compartments of the vagina of control and neonatally diethylstilbestrol-treated adult mice. J. Steroid Biochem. 35:617–621; 1990.

    Article  PubMed  CAS  Google Scholar 

  10. Galand, P.; Rodesch, F.; Leroy, F., et al. Radioautographic evaluation of the estrogen-dependent proliferative pool in the stem cell compartment of the mouse uterine and vaginal epithelia. Exp. Cell Res. 48:595–604; 1967.

    Article  PubMed  CAS  Google Scholar 

  11. Galand, P.; Leroy, F.; Chretien, J. Effect of oestradiol on cell proliferation and histological changes in the uterus and vagina of mice. J. Endocrinol. 49:243–252; 1971.

    PubMed  CAS  Google Scholar 

  12. Gilchrest, B. A.; Karassik, R. L.; Wilkins, L. M., et al. Autocrine and paracrine growth stimulation of cells derived from human skin. J. Cell. Physiol. 117:235–240; 1983.

    Article  PubMed  CAS  Google Scholar 

  13. Hinegardner, R. T. An improved fluorometric assay for DNA. Anal. Biochem. 39:197–201; 1971.

    Article  PubMed  CAS  Google Scholar 

  14. Iguchi, T.; Uchima, F.-D. A.; Ostrander, P. L., et al. Growth of normal mouse vaginal epithelial cells in and on collagen gels. Proc. Natl. Acad. Sci. USA 80:3743–3747; 1983.

    Article  PubMed  CAS  Google Scholar 

  15. Iguchi, T.; Uchima, F.-D. A.; Bern, H. A. Growth of mouse vaginal epithelial cells in culture: effect of sera and supplemented serum-free media. In Vitro Cell. Dev. Biol. 23:535–539; 1987.

    Article  PubMed  CAS  Google Scholar 

  16. Ikeda, T.; Sirbasku, D. A. Purification and properties of a mammary-uterine-pituitary tumor cell growth factor from pregnant sheep uterus. J. Biol. Chem. 259:4049–4061; 1984.

    PubMed  CAS  Google Scholar 

  17. Kirkland, J. L.; Gardner, R. M.; Ireland, J. S., et al. The effect of hypophysectomy on the uterine response to estradiol. Endocrinology 101:403–410; 1977.

    PubMed  CAS  Google Scholar 

  18. Knabbe, C.; Lippman, M. E.; Wakefield, L. M., et al. Evidence that transforming growth factor-β is a hormonally regulated negative growth factor in human breast cancer cells. Cell 48:417–428; 1987.

    Article  PubMed  CAS  Google Scholar 

  19. Kronenberg, M. S.; Clark, J. H. Changes in keratin expression during the estrogen-mediated differentiation of rat vaginal epithelium. Endocrinology 117:1480–1489; 1985.

    PubMed  CAS  Google Scholar 

  20. Licht, P.; McCreery, B. R.; Barnes, R., et al. Seasonal and stress related changes in plasma gonadotropins, sex steroids and corticosterone in the bullfrog,Rana catesbeiana. Gen. Comp. Endocrinol. 50:124–145; 1983.

    Article  PubMed  CAS  Google Scholar 

  21. Lin, T.-H.; Kirkland, J. L.; Mukku, V. R., et al. Regulation of deoxyribonucleic acid polymerase activity in uterine luminal epithelium after multiple doses of estrogen. Endocrinology 122:1403–1409; 1988.

    Article  PubMed  CAS  Google Scholar 

  22. MacLusky, N. J.; Naftolon, F.; Krey, L. C., et al. The catechol estrogens. J. Steroid Biochem. 15:111–124; 1981.

    Article  PubMed  CAS  Google Scholar 

  23. Martin, L.; Finn, C. A.; Trinder, G. Hypertrophy and hyperplasia in the mouse uterus after oestrogen treatment: an autoradiographic study. J. Endocrinol. 56:133–144; 1973.

    PubMed  CAS  Google Scholar 

  24. Seegers, J. C.; Aveling, M.-L.; van Aswegen, C. H., et al. The cytotoxic effects of estradiol-17β, catecholestradiols and methoxyestradiols on dividing MCF-7 and HeLa cells. J. Steroid Biochem. 32:797–809; 1989.

    Article  PubMed  CAS  Google Scholar 

  25. Sirbasku, D. A. Estrogen induction of growth factors specific for hormone-responsive mammary, pituitary, and kidney tumor cells. Proc. Natl. Acad. Sci. USA 75:3786–3790; 1978.

    Article  PubMed  CAS  Google Scholar 

  26. Snell, G. D. Reproduction. In: Snell, G. D., ed. Biology of the laboratory mouse. New York: Dover Publication; 1956:55–88.

    Google Scholar 

  27. Sonnenschein, C.; Pierce, G. B. Symposium on cell multiplication and cancer: positive or negative control. Cancer Res. 41:4742–4743; 1981.

    Google Scholar 

  28. Sonnenschein, C.; Olea, N.; Pasanen, M. E., et al. Negative controls of cell proliferation: human prostate cancer cells and androgens. Cancer Res. 49:3474–3481; 1989.

    PubMed  CAS  Google Scholar 

  29. Soto, A. M.; Sonnenschein, C. The role of estrogens on the proliferation of human breast tumor cells (MCF-7). J. Steroid Biochem. 23:87–94; 1985.

    Article  PubMed  CAS  Google Scholar 

  30. Soto, A. M.; Sonnenschein, C. Cell proliferation of estrogen sensitive cells: the case for negative control. Endocrin. Rev. 8:44–52; 1987.

    Article  CAS  Google Scholar 

  31. Soto, A. M.; Sonnenschein, C. Mechanism of estrogen action on cellular proliferation: evidence for indirect and negative control on cloned breast tumor cells. Biochem. Biophys. Res. Comm. 122:1097–1103; 1984.

    Article  PubMed  CAS  Google Scholar 

  32. Taguchi, O.; Bigsby, R. M.; Cunha, G. R. Estrogen responsiveness and the estrogen receptor during development of the murine female reproductive tract. Dev. Growth & Differ. 30:301–313; 1988.

    Article  CAS  Google Scholar 

  33. Uchima, F.-D. A.; Edery, M.; Mills, K. T., et al. Estrogen and progestin receptors in mouse vaginal epithelium and fibromuscular wall. Biochim. Biophys. Acta 841:135–138; 1985.

    PubMed  CAS  Google Scholar 

  34. Uchima, F.-D. A.; Edery, M.; Iguchi, T., et al. Growth of mouse vaginal epithelial cells in culture: functional integrity of the estrogen receptor system and failure of estrogen to induce proliferation. Cancer Lett. 17:115–123; 1987.

    Google Scholar 

  35. Vijayasaradhi, S.; Khar, A.; Gupta, P. D. Effect of steroid hormones on vaginal epithelial cells: an in vitro model for steroid hormone action. J. Biosci. 12:257–265; 1987.

    CAS  Google Scholar 

  36. Wood, E. J.; Raxworthy, M. J.; Holland, D. B. Retinoids and the epidermis. Biochem. Soc. Trans. 16:668–671; 1988.

    PubMed  CAS  Google Scholar 

  37. Yang, J.; Flynn, D.; Larson, L., et al. Growth in primary culture of mouse submandibular epithelial cells embedded in collagen gels. In Vitro 18:435–442; 1982.

    PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Francis-Dean A. Uchima

    Present address: Telios Pharmaceuticals, Inc., 2909 Science Park Rd., 92121, San Diego, CA

Authors and Affiliations

  1. Department of Integrative Biology and Cancer Research Laboratory, University of California, 94720, Berkeley, California

    Pei-San Tsai, Francis-Dean A. Uchima, Susan T. Hamamoto & Howard A. Bern

Authors
  1. Pei-San Tsai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Francis-Dean A. Uchima
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Susan T. Hamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Howard A. Bern
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by grants CA-05388 and CA-09041 from the National Institutes of Health, Bethesda, MD.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tsai, PS., Uchima, FD.A., Hamamoto, S.T. et al. Proliferation and differentiation of prepubertal mouse vaginal epithelial cells in vitro and the specificity of estrogen-induced growth retardation. In Vitro Cell Dev Biol - Animal 27, 461–468 (1991). https://doi.org/10.1007/BF02631145

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02631145

Key words

Search

Navigation