title: Functional proteomics approach to investigate the biological activities of cDNAs implicated in breast cancer.
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.
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston Massachusetts 02115, USA.
authors.forename: Abigail E
authors.initials: AE
authors.lastname: Witt
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Hines
authors.forename: Nicole L
authors.initials: NL
authors.lastname: Collins
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Hu
authors.forename: Ruwanthi N
authors.initials: RN
authors.lastname: Gunawardane
authors.forename: Donna
authors.initials: D
authors.lastname: Moreira
authors.forename: Jacob
authors.initials: J
authors.lastname: Raphael
authors.forename: Daniel
authors.initials: D
authors.lastname: Jepson
authors.forename: Malvika
authors.initials: M
authors.lastname: Koundinya
authors.forename: Andreas
authors.initials: A
authors.lastname: Rolfs
authors.forename: Barbara
authors.initials: B
authors.lastname: Taron
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Isakoff
authors.forename: Joan S
authors.initials: JS
authors.lastname: Brugge
authors.forename: Joshua
authors.initials: J
authors.lastname: LaBaer
mesh_terms: D001943:Breast Neoplasms / Q000737:chemistry / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D018076:DNA, Complementary / Q000302:isolation & purification*; D005240:Feasibility Studies; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D040901:Proteomics; D015398:Signal Transduction / Q000235:genetics
chemical_list: D018076:DNA, Complementary