Abstract

The specific aim of this proposed research is to develop and extensively characterize human mammary epithelial cells (HMEC) transformed in vitro by chemical carcinogens, as a means of understanding the number and nature of the events occurring during immortal and/or malignant transformation. The relationship of these events to expression of functional differentiation, involvement of known oncogenes, and response to growth controlling factors will be explored in order to suggest possible mechanisms contributing to the development of the immortal and/or malignant phenotype. We have already demonstrated that benzo(a)pyrene (BaP) can induce an extended life in culture as well as the immortal transformation of HMEC from at least one specimen donor and that the infection of immortal cells with the Kirsten mouse sarcoma virus (K-MSV) will render them tumorigenic in nude mice. We will continue these studies by (1) characterizing the extended life, immortally transformed, and malignantly transformed HMEC, for a variety of properties associated with malignant progression and/or state of functional differentiation. We will look for the presence or absence of coordinate expression of these different markers with each other, and with the growth properties of the various cell types. (2) Obtaining a larger number and variety of transformed cell types by exposing HMEC from at least two other individual donors to BaP, and by exposing the existing extended life and immortal cells to further chemical carcinogens and/or promoters. (3) Fusing combinations of immortal cell lines, extended life, and normal cells to determine dominance or recessiveness, and possible complementation groups, for immortality and abnormal properties. (4) Screening the various HMEC substrates for alteration or activation of known oncogenes, cellular processing of the p21 protein, and susceptibility to K-MSV induced malignant transformation. (5) Providing HMEC substrates to other investigators interested in applying their specific expertise to this system. It is hoped that this research will provide a well characterized human epithelial cell system for the study of the etiology and mechanisms of human cellular carcinogenesis.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA024844-07
Application #
3166592
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1979-09-01
Project End
1989-08-31
Budget Start
1985-09-01
Budget End
1986-08-31
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Lawrence Berkeley National Laboratory
Department
Type
Organized Research Units
DUNS #
078576738
City
Berkeley
State
CA
Country
United States
Zip Code
94720

  Publications

Garbe, James C; Vrba, Lukas; Sputova, Klara et al. (2014) Immortalization of normal human mammary epithelial cells in two steps by direct targeting of senescence barriers does not require gross genomic alterations. Cell Cycle 13:3423-35
Garbe, James C; Bhattacharya, Sanchita; Merchant, Batul et al. (2009) Molecular distinctions between stasis and telomere attrition senescence barriers shown by long-term culture of normal human mammary epithelial cells. Cancer Res 69:7557-68
Nijjar, Tarlochan; Bassett, Ekaterina; Garbe, James et al. (2005) Accumulation and altered localization of telomere-associated protein TRF2 in immortally transformed and tumor-derived human breast cells. Oncogene 24:3369-76
Stampfer, Martha R; Garbe, James; Nijjar, Tarlochan et al. (2003) Loss of p53 function accelerates acquisition of telomerase activity in indefinite lifespan human mammary epithelial cell lines. Oncogene 22:5238-51
Stampfer, Martha R; Yaswen, Paul (2003) Human epithelial cell immortalization as a step in carcinogenesis. Cancer Lett 194:199-208
Yaswen, P; Stampfer, M R (2001) Epigenetic changes accompanying human mammary epithelial cell immortalization. J Mammary Gland Biol Neoplasia 6:223-34
Stampfer, M R; Garbe, J; Levine, G et al. (2001) Expression of the telomerase catalytic subunit, hTERT, induces resistance to transforming growth factor beta growth inhibition in p16INK4A(-) human mammary epithelial cells. Proc Natl Acad Sci U S A 98:4498-503
Nonet, G H; Stampfer, M R; Chin, K et al. (2001) The ZNF217 gene amplified in breast cancers promotes immortalization of human mammary epithelial cells. Cancer Res 61:1250-4
Stampfer, M R; Yaswen, P (2000) Culture models of human mammary epithelial cell transformation. J Mammary Gland Biol Neoplasia 5:365-78
Kim, D W; Sovak, M A; Zanieski, G et al. (2000) Activation of NF-kappaB/Rel occurs early during neoplastic transformation of mammary cells. Carcinogenesis 21:871-9

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