Investigating the role of ADP-ribosylation factor 6 in tumor cell invasion and extracellular signal-regulated kinase activation.
Tumor cell invasion is a coordinated process involving the formation of invadopodia and the localized degradation of the extracellular matrix (ECM). The process of cell invasion is regulated by cell-signaling proteins such as Ras-related GTPases and members of the mitogen-activated protein kinase (MAPK) family. Our studies have focused on the role of the ADP-ribosylation factor 6 (ARF6) GTPase in the process of tumor cell invasion. Using activated and dominant negative mutants of ARF6 in a tumor cell culture model, our laboratory has demonstrated that the GTPase cycle of ARF6 regulates invadopodia formation and matrix degradation. Furthermore, ARF6-mediated cell invasion was found to be dependent on the activation of the extracellular signal-regulated kinase (ERK). These findings demonstrate a critical role for ARF6 in ERK activation and tumor cell invasion. To investigate the role of ARF6 in tumor cell invasion and ERK activation, a number of methods were employed. These procedures include transfection of LOX cells, in vitro matrix-degradation assays, immunofluorescence microscopy, and biochemical assays. These approaches can be applied effectively to measure the degree of invasiveness fostered by ARF6 and/or other GTPases and to examine the subcellular distribution of the molecular players that are trafficked or recruited to sites of cell invasion.
Hoover
Holly
H
Department of Biological Sciences, University of Notre Dame, Indiana, USA.
Muralidharan-Chari
Vandhana
V
Tague
Sarah
S
D'Souza-Schorey
Crislyn
C
D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000502:physiology*; D045744:Cell Line, Tumor; D004789:Enzyme Activation / Q000502:physiology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D009361:Neoplasm Invasiveness / Q000503:physiopathology*; D014162:Transfection
D000089982:ADP-Ribosylation Factor 6; D048049:Extracellular Signal-Regulated MAP Kinases; D020727:ADP-Ribosylation Factors; C000715348:ARF6 protein, human