PMID- 23302090 OWN - NLM STAT- MEDLINE DCOM- 20130614 LR - 20211021 IS - 1471-2407 (Electronic) IS - 1471-2407 (Linking) VI - 13 DP - 2013 Jan 9 TI - APC/beta-catenin-rich complexes at membrane protrusions regulate mammary tumor cell migration and mesenchymal morphology. PG - 12 LID - 10.1186/1471-2407-13-12 [doi] AB - BACKGROUND: The APC tumor suppressor is mutated or downregulated in many tumor types, and is prominently localized to punctate clusters at protrusion tips in migratory cells, such as in astrocytes where it has been implicated in directed cell motility. Although APC loss is considered an initiating event in colorectal cancer, for example, it is less clear what role APC plays in tumor cell motility and whether loss of APC might be an important promoter of tumor progression in addition to initiation. METHODS: The localization of APC and beta-catenin was analyzed in multiple cell lines, including non-transformed epithelial lines treated with a proteasome inhibitor or TGFbeta to induce an epithelial-to-mesenchymal transition (EMT), as well as several breast cancer lines, by immunofluorescence. APC expression was knocked down in 4T07 mammary tumor cells using lentiviral-mediated delivery of APC-specific short-hairpin (sh) RNAs, and assessed using quantitative (q) reverse-transcriptase (RT)-PCR and western blotting. Tumor cell motility was analyzed by performing wound-filling assays, and morphology via immunofluorescence (IF) and phase-contrast microscopy. Additionally, proliferation was measured using BrdU incorporation, and TCF reporter assays were performed to determine beta-catenin/TCF-mediated transcriptional activity. RESULTS: APC/beta-catenin-rich complexes were observed at protrusion ends of migratory epithelial cells treated with a proteasome inhibitor or when EMT has been induced and in tumor cells with a mesenchymal, spindle-like morphology. 4T07 tumor cells with reduced APC levels were significantly less motile and had a more rounded morphology; yet, they did not differ significantly in proliferation or beta-catenin/TCF transcriptional activity. Furthermore, we found that APC/beta-catenin-rich complexes at protrusion ends were dependent upon an intact microtubule cytoskeleton. CONCLUSIONS: These findings indicate that membrane protrusions with APC/beta-catenin-containing puncta control the migratory potential and mesenchymal morphology of mammary tumor cells and suggest that APC loss during later stages of tumor progression might impact tumor cell dissemination or colonization. FAU - Odenwald, Matthew A AU - Odenwald MA AD - Department of Surgery, University of Chicago, Chicago, IL 60637, USA. FAU - Prosperi, Jenifer R AU - Prosperi JR FAU - Goss, Kathleen H AU - Goss KH LA - eng GR - T32 HD007009/HD/NICHD NIH HHS/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20130109 PL - England TA - BMC Cancer JT - BMC cancer JID - 100967800 RN - 0 (APC protein, human) RN - 0 (Adenomatous Polyposis Coli Protein) RN - 0 (CTNNB1 protein, human) RN - 0 (CTNNB1 protein, mouse) RN - 0 (Multiprotein Complexes) RN - 0 (Proteasome Inhibitors) RN - 0 (Transforming Growth Factor beta) RN - 0 (beta Catenin) RN - EC 3.4.25.1 (Proteasome Endopeptidase Complex) SB - IM MH - Adenomatous Polyposis Coli Protein/genetics/*metabolism MH - Animals MH - Blotting, Western MH - Breast Neoplasms/genetics/*metabolism/pathology MH - *Cell Movement/drug effects MH - Cell Proliferation MH - *Cell Shape/drug effects MH - Cell Surface Extensions/drug effects/*metabolism/pathology MH - Cytoskeleton/metabolism MH - Dogs MH - *Epithelial-Mesenchymal Transition/drug effects MH - Female MH - Fluorescent Antibody Technique MH - Genes, Reporter MH - HCT116 Cells MH - Humans MH - Madin Darby Canine Kidney Cells MH - Mice MH - Microscopy, Phase-Contrast MH - Multiprotein Complexes MH - Neoplasm Invasiveness MH - Proteasome Endopeptidase Complex/drug effects/metabolism MH - Proteasome Inhibitors/pharmacology MH - RNA Interference MH - Reverse Transcriptase Polymerase Chain Reaction MH - Time Factors MH - Transfection MH - Transforming Growth Factor beta/metabolism MH - beta Catenin/*metabolism PMC - PMC3556124 EDAT- 2013/01/11 06:00 MHDA- 2013/06/15 06:00 PMCR- 2013/01/09 CRDT- 2013/01/11 06:00 PHST- 2012/06/22 00:00 [received] PHST- 2013/01/05 00:00 [accepted] PHST- 2013/01/11 06:00 [entrez] PHST- 2013/01/11 06:00 [pubmed] PHST- 2013/06/15 06:00 [medline] PHST- 2013/01/09 00:00 [pmc-release] AID - 1471-2407-13-12 [pii] AID - 10.1186/1471-2407-13-12 [doi] PST - epublish SO - BMC Cancer. 2013 Jan 9;13:12. doi: 10.1186/1471-2407-13-12.