Establishment of three human breast epithelial cell lines derived from carriers of the 999del5 BRCA2 icelandic founder mutation
Germ line mutations in BRCA1 and BRCA2 account for a large proportion of inherited breast and ovarian cancer. Both genes are involved in DNA repair by homologous recombination and are thought to play a vital role in maintaining genomic stability. A major drawback for long-term functional studies of BRCA in general and BRCA2 in particular has been a lack of representative human breast epithelial cell lines. In the present study, we have established three cell lines from two patients harboring the 999del5 germ line founder mutation in the BRCA2 gene. Primary cultures were established from cellular outgrowth of explanted tissue and subsequently transfected with a retroviral construct containing the HPV-16 E6 and E7 oncogenes. Paired cancer-derived and normal-derived cell lines were established from one patient referred to as BRCA2-999del5-2T and BRCA2-999del5-2N, respectively. In addition, one cell line was derived from cancer-associated normal tissue from another patient referred to as BRCA2-999del5-1N. All three cell lines showed characteristics of breast epithelial cells as evidenced by expression of breast epithelial specific cytokeratins. Cytogenetic analysis showed marked chromosomal instability with tetraploidy and frequent telomeric association. In conclusion, we have established three breast cpithelial cell lines from two patients carrying the BRCA2 Icelandic 999del5 founder mutation. These cell lines from the basis for further studies on carcinogenesis and malignant progression of breast cancer on a defined genetic background.
Key wordsbreast cancer cell lines immortalization BRCA2 mutation
Unable to display preview. Download preview PDF.
- Arason, A.; Jonasdottir, A.; Barkardottir, R. B.; Bergthorsson, J. T.; Teare, M. D.; Easton, D. F.; Egilsson, V. A population study of mutations and LOH at breast cancer gene loci in tumours from sister pairs: two recurrent mutations seem to account for all BRCA1/BRCA2 linked breast cancer in Iceland. J. Med. Genet. 35:446–449; 1998.PubMedCrossRefGoogle Scholar
- Briand, P.; Petersen, O. W.; Van Deurs, B. A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium. In Vitro Cell. Dev. Biol. 23A:181–188; 1987.Google Scholar
- Fanjul, M.; Hollande, E. Morphogenesis of “duct-like” structures in three-dimensional cultures of human cancerous pancreatic duct cells (Capan-1). In Vitro Cell. Dev. Biol. 29A:574–584; 1993.Google Scholar
- ISCN. An international system for human cytogenetic nomenclature. Mitelman, F. (Ed.). Basel: S. Karger, 1995.Google Scholar
- Samouelian, V.; Maugard, C. M.; Jolicoeur, M.; Bertrand, R.; Arcand, S. L.; Tonin, P. N.; Provencher, D. M.; Mes-Masson, A. M. Chemosensitivity and radiosensitivity profiles of four new human epithelial ovarian cancer cell lines exhibiting genetic alterations in BRCA2, TGFbeta-RII, KRAS2, TP53 and/or CDNK2A. Cancer Chemother. Pharmacol. 54:497–504; 2004.PubMedCrossRefGoogle Scholar