Functional proteomics approach to investigate the biological activities of cDNAs implicated in breast cancer

Abigail E. Witt, Lisa M. Hines, Nicole L. Collins, Yanhui Hu, Ruwanthi N. Gunawardane, Donna Moreira, Jacob Raphael, Daniel Jepson, Malvika Koundinya, Andreas Rolfs, Barbara Taron, Steven J. Isakoff, Joan S. Brugge, Joshua LaBaer

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

Functional proteomics approaches that comprehensively evaluate the biological activities of human cDNAs may provide novel insights into disease pathogenesis. To systematically investigate the functional activity of cDNAs that have been implicated in breast carcinogenesis, we generated a collection of cDNAs relevant to breast cancer, the Breast Cancer 1000 (BC1000), and conducted screens to identify proteins that induce phenotypic changes that resemble events which occur during tumor initiation and progression. Genes were selected for this set using bioinformatics and data mining tools that identify genes associated with breast cancer. Greater than 1000 cDNAs were assembled and sequence verified with high-throughput recombination-based cloning. To our knowledge, the BC1000 represents the first publicly available sequence-validated human disease gene collection. The functional activity of a subset of the BC1000 collection was evaluated in cell-based assays that monitor changes in cell proliferation, migration, and morphogenesis in MCF-10A mammary epithelial cells expressing a variant of ErbB2 that can be inducibly activated through dimerization. Using this approach, we identified many cDNAs, encoding diverse classes of cellular proteins, that displayed activity in one or more of the assays, thus providing insights into a large set of cellular proteins capable of inducing functional alterations associated with breast cancer development.

Original languageEnglish (US)
Pages (from-to)599-610
Number of pages12
JournalJournal of Proteome Research
Volume5
Issue number3
DOIs
StatePublished - Mar 1 2006
Externally publishedYes

Keywords

  • Acinar morphogenesis
  • BC1000
  • Breast cancer
  • Cell-based assay
  • Functional proteomics
  • High-throughput
  • Invasion
  • Migration
  • Proliferation
  • cDNA expression

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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  • Cite this

    Witt, A. E., Hines, L. M., Collins, N. L., Hu, Y., Gunawardane, R. N., Moreira, D., Raphael, J., Jepson, D., Koundinya, M., Rolfs, A., Taron, B., Isakoff, S. J., Brugge, J. S., & LaBaer, J. (2006). Functional proteomics approach to investigate the biological activities of cDNAs implicated in breast cancer. Journal of Proteome Research, 5(3), 599-610. https://doi.org/10.1021/pr050395r