Signaling through ShcA Is Required for Transforming Growth Factor (cid:2) - and Neu/ErbB-2-Induced Breast Cancer Cell Motility and Invasion (cid:1) †

Cooperation between the Neu/ErbB-2 and transforming growth factor (cid:1) (TGF- (cid:1) ) signaling pathways enhances the invasive and metastatic capabilities of breast cancer cells; however, the underlying mechanisms mediating this synergy have yet to be fully explained. We demonstrate that TGF- (cid:1) induces the migration and invasion of mammary tumor explants expressing an activated Neu/ErbB-2 receptor, which requires signaling from autophosphorylation sites located in the C terminus. A systematic analysis of mammary tumor explants expressing Neu/ErbB-2 add-back receptors that couple to distinct signaling molecules has mapped the syner- gistic effect of TGF- (cid:1) -induced motility and invasion to signals emanating from tyrosine residues 1226/1227 and 1253 of Neu/ErbB-2. Given that the ShcA adaptor protein is known to interact with Neu/ErbB-2 through these residues, we investigated the importance of this signaling molecule in TGF- (cid:1) -induced cell motility and invasion. The reduction of ShcA expression rendered cells expressing activated Neu/ErbB-2, or add-back receptors signaling specifically through tyrosines 1226/1227 or 1253, unresponsive to TGF- (cid:1) -induced motility and invasion. In addition, a dominant-negative form of ShcA, lacking its three known tyrosine phosphorylation sites, completely abrogates the TGF- (cid:1) -induced migration and invasion of breast cancer cells expressing activated Neu/ErbB-2. Our results implicate signaling through the ShcA adaptor as a key component in the synergistic interaction between these pathways. Biosciences). Statistical analysis. Statistical significance values ( P values) for tumor growth, migration, invasion, and BrdU proliferation assays were obtained by performing a two-sample unequal-variance Student’s t test. of a of mediating

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