TY - JOUR
T1 - Telomerase immortalization of human mammary epithelial cells derived from a BRCA2 mutation carrier
AU - Lewis, Cheryl M.
AU - Herbert, Brittney Shea
AU - Bu, Dawei
AU - Halloway, Shane
AU - Beck, Adam
AU - Shadeo, Ashleen
AU - Zhang, Cindy
AU - Ashfaq, Raheela
AU - Shay, Jerry W.
AU - Euhus, David M.
N1 - Funding Information:
The authors would like to thank Dr. Charles Sherr for kindly providing the plasmid for cdk4, Dr. Brenda Grimes for advice, and Gregory Thompson, Denise LaRue, and the Wan Lam Laboratory array CGH group for their excellent technical assistance. Grant Support: This work was supported by a generous gift from the Charles M. Solomon Foundation, Genome Canada/Genome British Columbia and by the Department of Defense Breast Cancer Research Program DAMD-17-00-1-0438, to B-S.H. while a postdoctoral fellow at UT-Southwestern, and the Indiana Genomics Initiative (INGEN). INGEN of Indiana University is supported in part by Lilly Endowment Inc.
PY - 2006/9
Y1 - 2006/9
N2 - 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.
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.
KW - BRCA2
KW - Cellular aging and senescence
KW - Human mammary epithelial cells
KW - Immortalization
KW - Telomerase
UR - http://www.scopus.com/inward/record.url?scp=33746704989&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33746704989&partnerID=8YFLogxK
U2 - 10.1007/s10549-006-9189-9
DO - 10.1007/s10549-006-9189-9
M3 - Article
C2 - 16541310
AN - SCOPUS:33746704989
SN - 0167-6806
VL - 99
SP - 103
EP - 115
JO - Breast Cancer Research and Treatment
JF - Breast Cancer Research and Treatment
IS - 1
ER -