Direct-current electrical field (DCEF) induces directional migration in many cell types by activating intracellular signaling pathways. However, the mechanisms coupling the extracellular electric stimulation to the intracellular signals remain largely unknown. In this study, we show that DCEF directs migration of HT-1080
fibrosarcoma cells to the cathode, stimulates generation of
hydrogen peroxide and
superoxide through the activation of
NADPH oxidase, induces
anode-facing cytoskeleton polarization, and activates ERK signaling. Subsequent studies demonstrate that the electrotaxis of HT-1080
fibrosarcoma cells is abolished by
NADPH oxidase inhibitor and overexpression of
manganese superoxide dismutase (MnSOD), an
enzyme that hydrolyzes
superoxide. In contrast, overexpression of catalases, which hydrolyze
hydrogen peroxide, does not affect electrotaxis. MnSOD overexpression also eliminates cytoskeleton polarization as well as the activation of AKT, ERKs, and p38. In contrast, under
catalase overexpression, the cytoskeleton still polarizes and p38 activation is affected. Finally, we show that inhibition of ERK activation also abolishes DCEF-induced directional migration and cytoskeleton polarization. Collectively, our results indicate that
superoxide plays critical roles in DCEF-induced directional migration of
fibrosarcoma cells, possibly by regulating the activation of ERKs. This study provides novel insights into the current understanding of DCEF-mediated
cancer cell directional migration and
metastasis.