Our long term objective is to induce preneoplastic and neoplastic transformation of human mammary epithelial cells, under defined in vitro conditions, with direct- and indirect-acting chemical carcinogens. An ability to consistently transform human mammary cells under defined in vitro conditions is essential to designing experiments to understand the cellular-molecular events underlying transformation and histogenesis of human breast cancer(s). Our strategy is to transform human epithelial cells grown under serum- free conditions in a collagen gel culture system, as well as in monolayer culture systems using various matrices. Specifically, we will: a) analyze the growth regulation of normal, benign (fibrocystic) and carcinomatous human mammary cells in collagen culture, under serum-free conditions in the presence of hormones (estrogen, progesterone, prolactin, etc.), growth factors (EGF, FGF, TGF alpha, TGF beta, etc.) and nonhormonal agents (lipids, TPA, Li+); b) use these endpoints to design culture conditions which facilitate human mammary epithelial cell transformation at high yield, in a manner similar to those developed for transformation of murine epithelial cells; c) use cDNA probes to determine whether amplification, rearrangement or expression of plasminogen activators (tPA, urokinase), TGF alpha, TGF beta, EGF receptor, glutathione-s-transferase-P, gamma-glutamyl transpeptidase and 0-6 alkyl-DNA-alkyltransferase are reliable, quantitative molecular markers for transformation and tumor histogenesis; d) determine whether changes at the molecular level correlate with phenotype changes (karyotypic, immunocytochemical, ultramicroscopic); e) develop suitable in vitro and in vivo assay systems to identify those molecular-cellular changes which represent preneoplastic and neoplastic (malignant) transformants. Our experience and approach suggest that development of a system(s) for transformation of human mammary epithelial cell in vitro is an achievable goal.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA049374-04
Application #
3193406
Study Section
Special Emphasis Panel (SRC (50))
Project Start
1989-07-01
Project End
1994-05-31
Budget Start
1992-06-01
Budget End
1993-05-31
Support Year
4
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of California Berkeley
Department
Type
Schools of Arts and Sciences
DUNS #
094878337
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Popnikolov, N; Yang, J; Liu, A et al. (2001) Reconstituted normal human breast in nude mice: effect of host pregnancy environment and human chorionic gonadotropin on proliferation. J Endocrinol 168:487-96
Nandi, S; Guzman, R C; Yang, J (1995) Hormones and mammary carcinogenesis in mice, rats, and humans: a unifying hypothesis. Proc Natl Acad Sci U S A 92:3650-7
Popnikolov, N K; Yang, J; Guzman, R C et al. (1995) In vivo growth stimulation of collagen gel embedded normal human and mouse primary mammary epithelial cells. J Cell Physiol 163:51-60
Popnikolov, N K; Yang, J; Guzman, R C et al. (1995) Reconstituted human normal breast in nude mice using collagen gel or Matrigel. Cell Biol Int 19:539-46
Yang, J; Guzman, R C; Popnikolov, N et al. (1994) Phenotypic characterization of collagen gel embedded primary human breast epithelial cells in athymic nude mice. Cancer Lett 81:117-27
Yang, J; Popnikolov, N K; Sakthivel, R et al. (1994) Human breast cancers respond to growth factors in vivo but not in vitro. Cancer Lett 85:13-21
Sakthivel, R; Hamdan, M; Yang, J et al. (1993) Effect of TGF-alpha on growth of normal human breast epithelial cells in serum-free primary culture using 3-dimensional collagen gels. Cell Biol Int 17:387-97
Hamdan, M A; Guzman, R C; Yang, J et al. (1992) Depletion of O6-alkylguanine-DNA alkyltransferase by O6-benzylguanine in three-dimensional collagen cultures of normal human breast epithelial cells. Carcinogenesis 13:1743-9