MOLECULAR CHARACTERIZATION OF BREAST CANCER CELL LINES BY A LOW-DENSITY MICROARRAY.
Françoise de Longueville*, Marc Lacroix*, Anna-Maria Barbuto, Vincent Bertholet, Dominique Gallo,
Denis Larsimont, Laurence Marcq, Nathalie Zammatteo, Sophie Boffe, Guy Leclercq and Jose Remacle
( *: equal contributions)
International Journal of Oncology (2005) 27, 881-892
We designed a low-density microarray carrying 132 DNA capture sequences
highly specific for genes known to be differentially expressed among breast tumors and BCC lines or
associated with specific tumor properties (cell-cycle alteration, proteolysis, adhesion, hormone
sensitivity, etc). We analyzed gene expression in 11 BCC lines among which 6 had already been extensively
studied (BT-474, Hs578T, MCF-7, MDA-MB-231, MDA-MB-453, T-47D) and 5 were still poorly characterized
(Evsa-T, IBEP-1, IBEP-2, IBEP-3, KPL-1). Some data obtained were verified or extended by real-time
polymerase chain reaction (real-time PCR), Northern-blotting, Western blotting, immunohistochemistry and
cell growth studies. Clustering analysis of the low-density microarray data allowed the sorting of BCC
lines into two classes and supported a major discriminatory role for ER-alpha, confirming data from
previous studies. A few genes that are highly and specifically expressed in one cell line were identified
such as MGB1 / SCGB2A2 (mammaglobin 1 / secretoglobin family 2A, member 2) in Evsa-T cells, and
PIP / GCDFP15
(prolactin-inducible protein / gross cystic disease fluid protein-15 ) in MDA-MB-453 BCC, suggesting an apocrine origin for these latter cells.
Two BCC lines (IBEP-1 and IBEP-3) that had been previously characterized as ER-alpha-negative, were
classified by the low-density microarray among ER-alpha-positive lines
(MCF-7, T-47D, IBEP-2, BT-474, KPL-1) and were indeed confirmed as receptor-positive (at both mRNA and
protein levels) and hormone-responsive cells. In conclusion, our results support the use
of a low-density microarray approach in cases where the cost and exhaustiveness of high-density
microarrays may constitute a drawback; for instance, in obtaining a rapid phenotype evaluation
in cell populations freshly isolated from breast tumors.
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To obtain additional information on the breast cancer cell lines used in this article, see also:
ESTABLISHMENT AND CHARACTERIZATION OF THREE NEW BREAST-CANCER CELL LINES (1998).
RELEVANCE OF BREAST CANCER CELL LINES AS MODELS FOR BREAST TUMOURS: AN UPDATE (2004).
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To obtain additional information on genes of interest in breast cancer, see also:
A LOW-DENSITY DNA MICROARRAY FOR ANALYSIS OF MARKERS IN BREAST CANCER (Review article) (2002).
STABLE PORTRAIT OF BREAST TUMORS DURING PROGRESSION. DATA FROM BIOLOGY, PATHOLOGY, AND GENETICS (2004).
The Eppendorf Human Breast Cancer Dual Chip gene list.
Affiliations:
Eppendorf Array Technologies (EAT), Namur, Belgique (Belgium) (F.d.-L., A.-M.B., V.B., L.M., N.Z., S.B.)
Laboratoire Jean-Claude Heuson de Cancérologie Mammaire, Institut Jules Bordet - Université Libre de Bruxelles (ULB), Bruxelles
(Brussels), Belgique (Belgium) (M.L., D.G., G.L.)
Service d'Anatomie Pathologique, Institut Jules Bordet, Université Libre de Bruxelles (ULB), Bruxelles (Brussels), Belgique (Belgium) (D.L.)
Laboratoire de Biochimie et de Biologie Cellulaire, Université de Namur, Namur, Belgique (Belgium) (J.R.)
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