Telomerase immortalization of human mammary epithelial cells derived from a BRCA2 mutation carrier.
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.
Hamon Center for Therapeutic Oncology Research and Department of Surgery, University of Texas Southwestern Medical Center, Dallas, TX 75390-9039, USA.
Cheryl M
CM
Lewis
Brittney-Shea
BS
Herbert
Dawei
D
Bu
Shane
S
Halloway
Adam
A
Beck
Ashleen
A
Shadeo
Cindy
C
Zhang
Raheela
R
Ashfaq
Jerry W
JW
Shay
David M
DM
Euhus
D000818:Animals; D001940:Breast / Q000201:enzymology* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D003094:Collagen / Q000494:pharmacology; D020552:Comet Assay; D004338:Drug Combinations; D004847:Epithelial Cells / Q000201:enzymology* / Q000378:metabolism; D024522:Genes, BRCA2; D006579:Heterozygote; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D009368:Neoplasm Transplantation; D011509:Proteoglycans / Q000494:pharmacology; D019098:Telomerase / Q000378:metabolism*; D016615:Telomere / Q000648:ultrastructure; D014407:Tumor Cells, Cultured
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen; D019098:Telomerase