Our results indicated that miR-133b level was down-regulated in the tissues and cells of ESCC, which promoted the proliferation, anoikis resistance, anchorage-independent growth and eventually led to ESCC invasion and metastasis via targeting EGFR and the downstream ITGB4/FAK/Grb2, AKT and ERK pathways. Here, ITGB4 is linked to esophageal squamous cell carcinoma.