activator protein 1 (ap-1) contributes to epcam-dependent breast cancer invasion激活蛋白1(ap-1)有助于epcam-dependent乳腺癌入侵.pdfVIP

Sankpal et al. Breast Cancer Research 2011, 13:R124 /content/13/6/R124 RESEARCH ARTICLE Open Access Activator protein 1 (AP-1) contributes to EpCAM- dependent breast cancer invasion 1 1 1 1,2 1,2* Narendra V Sankpal , John D Mayfield , Mike W Willman , Timothy P Fleming and William E Gillanders Abstract Introduction: EpCAM is a cell-surface glycoprotein that is overexpressed in the majority of epithelial carcinomas. However, the functional role of EpCAM in regulating cancer invasion remains controversial, and the mechanism(s) underlying EpCAM-mediated regulation of breast cancer invasion remain to be defined. Methods: EpCAM expression was manipulated in breast cancer cell lines using RNA interference and cDNA expression constructs. Recombinant EpCAM was used to rescue EpCAM signaling following specific ablation of EpCAM. Protein and gene expression, invasion, transcription factor activity, and protein phosphorylation were measured using standard molecular biology techniques. Results: In loss-of-function, and gain-of-function experiments we demonstrate that EpCAM expression is associated with increased breast cancer invasion in vitro and in vivo. We demonstrate further that specific ablation of EpCAM expression is associated with decreased activator protein-1 (AP-1) transcription factor activity. Phosphoprotein analyses confirm that specific ablation of EpCAM is associated with decreased phosphorylation of the AP-1 subunit c-Jun. Recombinant soluble extracellular EpCAM (rEpCAM) is able to rescue invasion, AP-1 transcription factor activity, and c-Jun phosphorylation in a dose-dependent fashion. Pharmacologic inhibitors, and constitutively active constructs of the c-Jun N-te