Journal
JOURNAL OF PROTEOME RESEARCH
Volume 5, Issue 3, Pages 599-610Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr050395r
Keywords
functional proteomics; breast cancer; cell-based assay; cDNA expression; proliferation; migration; invasion; acinar morphogenesis; BC1000; high-throughput
Categories
Funding
- NCI NIH HHS [P01 CA080111-109001, P01 CA080111-079001, CA080111, P01 CA080111-099001, P01 CA080111-069001, P01 CA080111, CA089393, CA099191, P01 CA080111-089001] Funding Source: Medline
- NIGMS NIH HHS [U54 GM64346-04] Funding Source: Medline
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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.
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