title: Gbetagamma signaling promotes breast cancer cell migration and invasion.
abstract: Signaling through G protein-coupled receptors (GPCRs) promotes breast cancer metastasis. G proteins convey GPCR signals by dissociating into Galpha and Gbetagamma subunits. The aim of the present study was to determine whether blockade of Gbetagamma signaling suppresses breast cancer cell migration and invasion, which are critical components of metastasis. Conditioned media (CM) of NIH-3T3 fibroblasts are widely used as chemoattractants in in vitro cancer metastasis studies. Expression of a Gbetagamma scavenger peptide attenuated NIH-3T3 CM-induced migration and invasion of both metastatic breast cancer MDA-MB-231 and MDA-MB-436 cells by 40 to 50% without effects on cell viability. Migration and invasion of cells in response to NIH-3T3 CM were also blocked by 8-(4,5,6-trihydroxy-3-oxo-3H-xanthen-9-yl)-1-naph-thalene-carboxylic acid) (M119K), a Gbetagamma inhibitor, with maximum inhibition exceeding 80% and half-maximal inhibitory concentration (IC50) values of 1 to 2 microM. M119K also attenuated Rac-dependent formation of lamellipodia, a key structure required for metastasis. Constitutively active Rac1 rescued Gbetagamma blockade-mediated inhibition of breast cancer cell migration, whereas dominant negative Rac1 inhibited cell migration similar to Gbetagamma blockade. Furthermore, M119K suppressed Gi protein-coupled CXC chemokine receptor 4 (CXCR4)-dependent MDA-MB-231 cell migration by 80% with an IC50 value of 1 microM, whereas tyrosine kinase receptor-dependent cell migration was significantly less inhibited. However, CXCR4-dependent inhibition of adenylyl cyclase, a Gialpha-mediated response in MDA-MB-231 cells, was not blocked by M119K but was blocked by pertussis toxin, which selectively inactivates Gialpha. This report is the first to directly demonstrate the role of Gbetagamma in cancer cell migration and invasion and suggests that targeting Gbetagamma signaling pathways may provide a novel strategy for suppressing breast cancer metastasis.
authors.affiliation: Department of Pharmacology, Creighton University School of Medicine, Omaha, Nebraska 68178, USA.
authors.forename: Joseph K
authors.initials: JK
authors.lastname: Kirui
authors.forename: Yan
authors.initials: Y
authors.lastname: Xie
authors.forename: Dennis W
authors.initials: DW
authors.lastname: Wolff
authors.forename: Haihong
authors.initials: H
authors.lastname: Jiang
authors.forename: Peter W
authors.initials: PW
authors.lastname: Abel
authors.forename: Yaping
authors.initials: Y
authors.lastname: Tu
mesh_terms: D000262:Adenylyl Cyclases / Q000187:drug effects; D001943:Breast Neoplasms / Q000503:physiopathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology; D003510:Cyclohexanes / Q000494:pharmacology; D005260:Female; D044387:GTP-Binding Protein beta Subunits / Q000494:pharmacology*; D044388:GTP-Binding Protein gamma Subunits / Q000494:pharmacology*; D006801:Humans; D008856:Microscopy, Fluorescence; D009361:Neoplasm Invasiveness / Q000503:physiopathology*; D009362:Neoplasm Metastasis / Q000188:drug therapy / Q000503:physiopathology; D010446:Peptide Fragments / Q000502:physiology; D011554:Pseudopodia / Q000187:drug effects; D019718:Receptors, CXCR4 / Q000502:physiology; D011994:Recombinant Proteins; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014966:Xanthenes / Q000494:pharmacology; D020744:rac GTP-Binding Proteins / Q000187:drug effects / Q000502:physiology
chemical_list: C527276:CXCR4 protein, human; D003510:Cyclohexanes; C118981:G-protein Beta gamma; D044387:GTP-Binding Protein beta Subunits; D044388:GTP-Binding Protein gamma Subunits; C509953:M119 compound; D010446:Peptide Fragments; D019718:Receptors, CXCR4; D011994:Recombinant Proteins; D014966:Xanthenes; D017868:Cyclic AMP-Dependent Protein Kinases; C114043:BARKct protein, recombinant; D020744:rac GTP-Binding Proteins; D000262:Adenylyl Cyclases