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. 1977 Jun 1;73(3):561–577. doi: 10.1083/jcb.73.3.561

Mitogenic activity of pituitary hormones on cell cultures of normal and carcinogen-induced tumor epithelium from rat mammary glands

PS Rudland, RC Hallowes, H Durbin, D Lewis
PMCID: PMC2111412  PMID: 873989

Abstract

Cell suspension containing normal or tumor epithelium were readily obtained by enzymatically digesting rat mammary glands from perphenazine-treated (prolactin-hypersecreting) cycling, female virgin animals or hormone- responsive mammary tumors from animal treated with dimethylbenzanthracene. Cell suspensions were fractioned into predominantly epithelial and predominantly stromal cells by their differential rates of attachment to culture dishes. Both normal mammary and tumor epithelial cells were characterized by the presence of specific cell-junctional complexes, desmosome-like structures, surface microvilli, and their ability to synthesize casein. Serum-dependent protease activity was greater in cultures derived from tumors, and cells from such cultures grew in agarose whereas those from the non-neoplastic gland did not. The addition of prolactin to the culture medium stimulated DNA synthesis in primary or secondary epithelial cultures from tumors, whereas additional insulin and hydrocortisone with prolactin were required for similar levels of DNA synthesis in cultures from non-neoplastic glands. The fraction of cells synthesizing DNA was, however, smaller than that with 10 percent serum measured in the same time period. Both growth hormone and epidermal growth factor stimulated DNA synthesis but to a lesser extent than did prolactin. Prolactin with hydrocortisone and insulin were relatively inactive in promoting DNA synthesis of the nonepithelial cells whereas pituitary fibroblast growth factor was more active. These mitogenic effects were obtained when the hormones were added to the medium at near physiological concentrations, and paralleled the known activities of the hormones in control of mammary gland growth and development in the rat.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amenomori Y., Chen C. L., Meites J. Serum prolactin levels in rats during different reproductive states. Endocrinology. 1970 Mar;86(3):506–510. doi: 10.1210/endo-86-3-506. [DOI] [PubMed] [Google Scholar]
  2. COHEN S. Isolation of a mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the new-born animal. J Biol Chem. 1962 May;237:1555–1562. [PubMed] [Google Scholar]
  3. Cohen L. A., Tsuang J., Chan P. C. Characteristics of rat normal mammary epithelial cells and dimethylbenzanthracene-induced mammary adenocarcinoma cells grown in monolayer culture. In Vitro. 1974 Jul-Aug;10:51–62. doi: 10.1007/BF02615338. [DOI] [PubMed] [Google Scholar]
  4. Dilley W. G. Relationship of mitosis to alveolar development in the rat mammary gland. J Endocrinol. 1971 Jul;50(3):501–505. doi: 10.1677/joe.0.0500501. [DOI] [PubMed] [Google Scholar]
  5. Feldman M. K., Ceriani R. L. A comparative immunologic and electrophoretic analysis of rat and mouse caseins. Comp Biochem Physiol. 1970 Dec 1;37(3):421–427. doi: 10.1016/0010-406x(70)90570-0. [DOI] [PubMed] [Google Scholar]
  6. Gilbert S. F., Migeon B. R. D-valine as a selective agent for normal human and rodent epithelial cells in culture. Cell. 1975 May;5(1):11–17. doi: 10.1016/0092-8674(75)90086-0. [DOI] [PubMed] [Google Scholar]
  7. Gospodarowicz D., Jones K. L., Sato G. Purification of a growth factor for ovarian cells from bovine pituitary glands. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2295–2299. doi: 10.1073/pnas.71.6.2295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HUGGINS C., BRIZIARELLI G., SUTTON H., Jr Rapid induction of mammary carcinoma in the rat and the influence of hormones on the tumors. J Exp Med. 1959 Jan 1;109(1):25–42. doi: 10.1084/jem.109.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hallowes R. C., Wang D. Y., Lewis D. J. The lactogenic effects of prolactin and growth hormone on mammary gland explants from virgin and pregnant Sprague-Dawley rats. J Endocrinol. 1973 May;57(2):253–264. doi: 10.1677/joe.0.0570253. [DOI] [PubMed] [Google Scholar]
  10. Holley R. W., Kiernan J. A. Control of the initiation of DNA synthesis in 3T3 cells: low-molecular weight nutrients. Proc Natl Acad Sci U S A. 1974 Aug;71(8):2942–2945. doi: 10.1073/pnas.71.8.2942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kamely D., Rudland P. Nutrient-dependent arrest of fibroblast growth is partially reversed by insulin but not fibroblast growth factor. Nature. 1976 Mar 4;260(5546):51–53. doi: 10.1038/260051a0. [DOI] [PubMed] [Google Scholar]
  12. LYONS W. R., LI C. H., JOHNSON R. E. The hormonal control of mammary growth and lactation. Recent Prog Horm Res. 1958;14:219–254. [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  15. Lewis D., Hallowes R. C. Correlation between the effects of hormones on the synthesis of DNA in explants from induced rat mammary tumours and the growth of the tumours. J Endocrinol. 1974 Aug;62(2):225–240. doi: 10.1677/joe.0.0620225. [DOI] [PubMed] [Google Scholar]
  16. Mayne R., Barry J. M. Biochemical changes during development of mouse mammary tissue in organ culture. J Endocrinol. 1970 Jan;46(1):61–70. doi: 10.1677/joe.0.0460061. [DOI] [PubMed] [Google Scholar]
  17. NANDI S. Endocrine control of mammarygland development and function in the C3H/ He Crgl mouse. J Natl Cancer Inst. 1958 Dec;21(6):1039–1063. [PubMed] [Google Scholar]
  18. Oka T., Topper Y. J. Is prolactin mitogenic for mammary epithelium? Proc Natl Acad Sci U S A. 1972 Jul;69(7):1693–1696. doi: 10.1073/pnas.69.7.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pollack R., Risser R., Conlon S., Rifkin D. Plasminogen activator production accompanies loss of anchorage regulation in transformation of primary rat embryo cells by simian virus 40. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4792–4796. doi: 10.1073/pnas.71.12.4792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rudland P. S., Eckhart W., Gospodarowicz D., Seifert W. Cell transformation mutants are not susceptible to growth activation by fibroblast growth factor at permissive temperatures. Nature. 1974 Jul 26;250(464):337–339. doi: 10.1038/250337a0. [DOI] [PubMed] [Google Scholar]
  21. Rudland P. S., Pearlstein E., Kamely D., Nutt M., Eckhart W. Independent regulation of cellular properties in thermosensitive transformation mutants of mouse fibroblasts. Nature. 1975 Jul 3;256(5512):43–46. doi: 10.1038/256043a0. [DOI] [PubMed] [Google Scholar]
  22. Rudland P. S., Seeley M., Seifert W. Cyclic GMP and cyclic AMP levels in normal and transformed fibroblasts. Nature. 1974 Oct 4;251(5474):417–419. doi: 10.1038/251417a0. [DOI] [PubMed] [Google Scholar]
  23. Shin S. I., Freedman V. H., Risser R., Pollack R. Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4435–4439. doi: 10.1073/pnas.72.11.4435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stockdale F. E., Topper Y. J. The role of DNA synthesis and mitosis in hormone-dependent differentiation. Proc Natl Acad Sci U S A. 1966 Oct;56(4):1283–1289. doi: 10.1073/pnas.56.4.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. TALWALKER P. K., MEITES J. MAMMARY LOBULO-ALVEOLAR GROWTH IN ADRENO-OVARIECTOMIZED RATS FOLLOWING TRANSPLANTATION OF "MAMMOTROPIC" PITUITARY TUMOR. Proc Soc Exp Biol Med. 1964 Oct;117:121–124. doi: 10.3181/00379727-117-29512. [DOI] [PubMed] [Google Scholar]
  26. TALWALKER P. K., MEITES J. Mammary lobulo-alveolar growth induced by anterior pituitary hormones in adrenoovariectomized and adreno-ovariectomized-hypophysectomized rats. Proc Soc Exp Biol Med. 1961 Aug-Sep;107:880–883. doi: 10.3181/00379727-107-26783. [DOI] [PubMed] [Google Scholar]
  27. Thrash C. R., Cunningham D. D. Stimulation of division of density inhibited fibroblasts by glucocorticoids. Nature. 1973 Apr 6;242(5397):399–401. doi: 10.1038/242399a0. [DOI] [PubMed] [Google Scholar]
  28. Turkington R. W., Lockwood D. H., Topper Y. J. The induction of milk protein synthesis in post-mitotic mammary epithelial cells exposed to prolactin. Biochim Biophys Acta. 1967 Nov 28;148(2):475–480. doi: 10.1016/0304-4165(67)90144-4. [DOI] [PubMed] [Google Scholar]
  29. Turkington R. W., Majumder G. C., Kadoama N., MacIndoe J. H., Frantz W. L. Hormonal regulation of gene expression in mammary cells. Recent Prog Horm Res. 1973;29:417–455. doi: 10.1016/b978-0-12-571129-6.50015-4. [DOI] [PubMed] [Google Scholar]
  30. Welsch C. W., Rivera E. M. Differential effects of estrogen and prolactin on DNA synthesis in organ cultures of DMBA-induced rat mammary carcinoma. Proc Soc Exp Biol Med. 1972 Feb;139(2):623–626. doi: 10.3181/00379727-139-36201. [DOI] [PubMed] [Google Scholar]

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