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...

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Veröffentlicht in:In vitro cellular & developmental biology. Animal 2005-11, Vol.41 (10), p.337-342
Hauptverfasser: RUBNER FRIDRIKSDOTTIR, AGLA J, GUDJONSSON, THORARINN, HALLDORSSON, THORHALLUR, BJÖRNSSON, JOHANNES, STEINARSDOTTIR, MARGRET, JOHANNSSON, OSKAR THOR, ÖGMUNDSDOTTIR, HELGA M
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container_issue 10
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container_title In vitro cellular & developmental biology. Animal
container_volume 41
creator RUBNER FRIDRIKSDOTTIR, AGLA J
GUDJONSSON, THORARINN
HALLDORSSON, THORHALLUR
BJÖRNSSON, JOHANNES
STEINARSDOTTIR, MARGRET
JOHANNSSON, OSKAR THOR
ÖGMUNDSDOTTIR, HELGA M
description 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 associations. In conclusion, we have established three breast epithelial cell lines from two patients carrying the BRCA2 Icelandic 999del5 founder mutation. These cell lines form the basis for further studies on carcinogenesis and malignant progression of breast cancer on a defined genetic background.
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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 associations. In conclusion, we have established three breast epithelial cell lines from two patients carrying the BRCA2 Icelandic 999del5 founder mutation. 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subjects BRCA2 mutation
Breast - cytology
Breast cancer
CELL AND TISSUE MODELS
Cell Line, Tumor
Cell lines
Chromosomal Instability - genetics
Cultured cells
Cytogenetic Analysis
DNA Mutational Analysis
DNA Primers
Epithelial cells
Epithelial Cells - cytology
Exons
Female
Genes, BRCA2
Genetic mutation
Genetic Vectors - genetics
Humans
Iceland
immortalization
Immunohistochemistry
Karyotyping
Keratins - metabolism
Mutation - genetics
Oncogenes - genetics
Retroviridae
Stem cells
Tumor cell line
Tumor stem cells
Tumors
title ESTABLISHMENT OF THREE HUMAN BREAST EPITHELIAL CELL LINES DERIVED FROM CARRIERS OF THE 999del5 BRCA2 ICELANDIC FOUNDER MUTATION
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