Here, we showed that antcin-H-mediated inhibition of FAK-Src-paxillin signaling axis and RCC cell migration was accompanied by decrease of lamellipodium, indicating that disruption of the lamellipodium formation as well as inhibition of migration and wound closure attributed to reducing formation of focal adhesion and actin bundles through suppressing FAK-Src-paxillin signaling pathway in antcin-H-treated cells. This evidence concerns the gene PTK2 and renal cell carcinoma.