Telomerase immortalization of human mammary epithelial cells derived from a BRCA2 mutation carrier

Cheryl M. Lewis, Brittney Shea Herbert, Dawei Bu, Shane Halloway, Adam Beck, Ashleen Shadeo, Cindy Zhang, Raheela Ashfaq, Jerry W. Shay, David M Euhus

Research output: Contribution to journalArticle

Abstract

A novel human mammary epithelial cell line, HME348, was established from benign breast tissue from a 44-year-old germ-line BRCA2 mutation carrier with a history of stage 1 breast cancer. Mutation analysis showed that the patient had a known 6872del4 BRCA2 heterozygous mutation. The human mammary epithelial cells passaged in culture exhibited cellular replicative aging as evidenced by telomere shortening, lack of telomerase activity, and senescence. Ectopic expression of telomerase (hTERT) reconstituted telomerase activity in these cells and led to the immortalization of the cells. When grown on glass, the majority of immortalized HME348 cells expressed ESA and p63 with a small population also expressing EMA. In three-dimensional Matrigel® culture, HME348 cells formed complex branching acini structures that expressed luminal (EMA, CK18) and myoepithelial (p63, CALLA, CK14) markers. Three clones derived from this culture were also p63+ /ESA+ /EMA+/- on glass but formed similar acinar structures with both luminal and myoepithelial cell differentiation in Matrigel® confirming the mammary progenitor nature of these cells. Additionally, the experimentally immortalized HME348 cells formed acini in cleared mammary fat pads in vivo. As this is the first report establishing and characterizing a benign human mammary epithelial cell line derived from a BRCA2 patient without the use of viral oncogenes, these cells may be useful for the study of BRCA2 function in breast morphogenesis and carcinogenesis.

Original languageEnglish (US)
Pages (from-to)103-115
Number of pages13
JournalBreast Cancer Research and Treatment
Volume99
Issue number1
DOIs
StatePublished - Sep 2006
Externally publishedYes

Fingerprint

Telomerase
Breast
Epithelial Cells
Mutation
Glass
Telomere Shortening
Cell Line
Germ-Line Mutation
Acinar Cells
Cell Aging
Morphogenesis
Oncogenes
Adipose Tissue
Cell Differentiation
Carcinogenesis
Stem Cells
Clone Cells
Breast Neoplasms
Population

Keywords

  • BRCA2
  • Cellular aging and senescence
  • Human mammary epithelial cells
  • Immortalization
  • Telomerase

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Telomerase immortalization of human mammary epithelial cells derived from a BRCA2 mutation carrier. / Lewis, Cheryl M.; Herbert, Brittney Shea; Bu, Dawei; Halloway, Shane; Beck, Adam; Shadeo, Ashleen; Zhang, Cindy; Ashfaq, Raheela; Shay, Jerry W.; Euhus, David M.

In: Breast Cancer Research and Treatment, Vol. 99, No. 1, 09.2006, p. 103-115.

Research output: Contribution to journalArticle

Lewis, CM, Herbert, BS, Bu, D, Halloway, S, Beck, A, Shadeo, A, Zhang, C, Ashfaq, R, Shay, JW & Euhus, DM 2006, 'Telomerase immortalization of human mammary epithelial cells derived from a BRCA2 mutation carrier', Breast Cancer Research and Treatment, vol. 99, no. 1, pp. 103-115. https://doi.org/10.1007/s10549-006-9189-9
Lewis, Cheryl M. ; Herbert, Brittney Shea ; Bu, Dawei ; Halloway, Shane ; Beck, Adam ; Shadeo, Ashleen ; Zhang, Cindy ; Ashfaq, Raheela ; Shay, Jerry W. ; Euhus, David M. / Telomerase immortalization of human mammary epithelial cells derived from a BRCA2 mutation carrier. In: Breast Cancer Research and Treatment. 2006 ; Vol. 99, No. 1. pp. 103-115.
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AB - A novel human mammary epithelial cell line, HME348, was established from benign breast tissue from a 44-year-old germ-line BRCA2 mutation carrier with a history of stage 1 breast cancer. Mutation analysis showed that the patient had a known 6872del4 BRCA2 heterozygous mutation. The human mammary epithelial cells passaged in culture exhibited cellular replicative aging as evidenced by telomere shortening, lack of telomerase activity, and senescence. Ectopic expression of telomerase (hTERT) reconstituted telomerase activity in these cells and led to the immortalization of the cells. When grown on glass, the majority of immortalized HME348 cells expressed ESA and p63 with a small population also expressing EMA. In three-dimensional Matrigel® culture, HME348 cells formed complex branching acini structures that expressed luminal (EMA, CK18) and myoepithelial (p63, CALLA, CK14) markers. Three clones derived from this culture were also p63+ /ESA+ /EMA+/- on glass but formed similar acinar structures with both luminal and myoepithelial cell differentiation in Matrigel® confirming the mammary progenitor nature of these cells. Additionally, the experimentally immortalized HME348 cells formed acini in cleared mammary fat pads in vivo. As this is the first report establishing and characterizing a benign human mammary epithelial cell line derived from a BRCA2 patient without the use of viral oncogenes, these cells may be useful for the study of BRCA2 function in breast morphogenesis and carcinogenesis.

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