The present study showed for the first time that antcin-H, a steroid-like compound isolated from a famous anticancer medicinal mushroom A. cinnamomea, inhibited Src, FAK, and ERK1/2 signaling pathways and thereby decreased phosphorylated-paxillin, phosphorylated-C/EBP-β, and total c-Fos levels and downregulated vimentin and MMPs expression, finally leading to impaired lamellipodium formation and cellular migration/invasion at nontoxic concentrations in human RCC 786-0 cells. Here, PTK2 is linked to renal cell carcinoma.