ABSTRACTAberrant overexpression of autocrine motility factor and its receptor (AMFR) is observed in many cancers but not in esophageal squamous cell cancer (ESCC). In this study, upregulated rho-associated protein kinase 2, p-cofilin and intracellular adhesion molecule-1, and downregulated E-cadherin were found in ESCC cells transfected with the plasmid pcDNA 3.1-AMFR-C, while opposite results were observed in ESCC cells transfected with siRNA against AMFR. Additionally, an elevated invasion of ESCC cells by AMFR was reversed by rho-associated protein kinase 2 inhibitor Y-27632, suggesting that AMFR pathway promotes invasion of ESCC cells and may be a potential target for ESCC therapy.