Molecular characterization of breast cancer cell lines by a low-density microarray

Int J Oncol. 2005 Oct;27(4):881-92.

Abstract

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 (mammaglobin 1) in Evsa-T cells, and PIP (prolactin-inducible protein) 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 utility 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotinylation
  • Blotting, Northern
  • Blotting, Western
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / metabolism*
  • Breast Neoplasms / pathology
  • Cell Adhesion
  • Cell Line, Tumor
  • Cell Proliferation
  • Cluster Analysis
  • DNA, Complementary / metabolism
  • Estrogen Receptor alpha / metabolism
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Image Processing, Computer-Assisted
  • Immunohistochemistry
  • Mammaglobin A
  • Neoplasm Proteins / metabolism
  • Nucleic Acid Hybridization
  • Oligonucleotide Array Sequence Analysis*
  • Phenotype
  • Polymerase Chain Reaction
  • RNA / metabolism
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Uteroglobin / metabolism

Substances

  • DNA, Complementary
  • Estrogen Receptor alpha
  • Mammaglobin A
  • Neoplasm Proteins
  • RNA, Messenger
  • SCGB2A2 protein, human
  • RNA
  • Uteroglobin