A cell motility screen reveals role for MARCKS-related protein in adherens junction formation and tumorigenesis.
Invasion through the extracellular matrix (ECM) is important for wound healing, immunological responses and metastasis. We established an invasion-based cell motility screen using Boyden chambers overlaid with Matrigel to select for pro-invasive genes. By this method we identified antisense to MARCKS related protein (MRP), whose family member MARCKS is a target of miR-21, a microRNA involved in tumor growth, invasion and metastasis in multiple human cancers. We confirmed that targeted knockdown of MRP, in both EpRas mammary epithelial cells and PC3 prostate cancer cells, promoted in vitro cell migration that was blocked by trifluoperazine. Additionally, we observed increased immunofluoresence of E-cadherin, beta-catenin and APC at sites of cell-cell contact in EpRas cells with MRP knockdown suggesting formation of adherens junctions. By wound healing assay we observed that reduced MRP supported collective cell migration, a type of cell movement where adherens junctions are maintained. However, destabilized adherens junctions, like those seen in EpRas cells, are frequently important for oncogenic signaling. Consequently, knockdown of MRP in EpRas caused loss of tumorigenesis in vivo, and reduced Wnt3a induced TCF reporter signaling in vitro. Together our data suggest that reducing MRP expression promotes formation of adherens junctions in EpRas cells, allowing collective cell migration, but interferes with oncogenic beta-catenin signaling and tumorigenesis.
Finlayson
Alexander E
AE
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Freeman
Kevin W
KW
D022005:Adherens Junctions / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism; D002148:Calmodulin-Binding Proteins; D002450:Cell Communication; D045744:Cell Line, Tumor; D002465:Cell Movement; D002471:Cell Transformation, Neoplastic; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D035683:MicroRNAs / Q000378:metabolism; D008840:Microfilament Proteins; D008856:Microscopy, Fluorescence / Q000379:methods; D009362:Neoplasm Metastasis; D015398:Signal Transduction; D014945:Wound Healing; D051176:beta Catenin / Q000378:metabolism
D015820:Cadherins; D002148:Calmodulin-Binding Proteins; C070177:MARCKSL1 protein, human; C524940:MIRN21 microRNA, human; D008565:Membrane Proteins; D035683:MicroRNAs; D008840:Microfilament Proteins; D051176:beta Catenin