Acquisition of the metastatic phenotype is accompanied by H2O2-dependent activation of the p130Cas signaling complex.
Reactive oxygen species (ROS) have emerged as cellular signaling molecules and are implicated in metastatic disease by their ability to drive invasion and migration. Here, we define the signaling adaptor protein p130Cas (Crk-associated substrate) as a key redox-responsive molecular trigger that is engaged in highly invasive metastatic bladder tumor cell lines. Endogenous shifts in steady-state hydrogen peroxide (H2O2) that accompany the metastatic phenotype increase p130Cas phosphorylation, membrane recruitment and association with the scaffolding protein Crk, and subsequent Rac1 activation and actin reorganization. Both enzymatic and nonenzymatic scavenging of H2O2 abrogates p130Cas-dependent signaling and the migratory and invasive activity of the metastatic bladder tumor cells. Disruption of p130Cas attenuates both invasion and migration of the metastatic variant (253J-BV). 253J-BV cells displayed an increase in global thiol oxidation and a concomitant decrease in total phosphatase activity, common target proteins of active-site cysteine oxidation. The dependence of phosphatases on regulation of p130Cas was highlighted when depletion of PTPN12 enhanced p130cas phosphorylation and the migratory behavior of a noninvasive parental bladder tumor control (253J). These data show that the metastatic phenotype is accompanied by increases in steady-state H2O2 production that drive promigratory signaling and suggest that antioxidant-based therapeutics may prove useful in limiting bladder tumor invasiveness.
College of Nanoscale Sciences and Engineering, University at Albany, SUNY, Albany, NY 12203, USA.
Nadine
N
Hempel
Toni R
TR
Bartling
Badar
B
Mian
J Andres
JA
Melendez
D000199:Actins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D050719:Crk-Associated Substrate Protein / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D009361:Neoplasm Invasiveness / Q000235:genetics* / Q000473:pathology; D009362:Neoplasm Metastasis; D010766:Phosphorylation; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000473:pathology
D000199:Actins; C493841:BCAR1 protein, human; D050719:Crk-Associated Substrate Protein; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; C516738:PTPN12 protein, human; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12