High mobility group box 1 (HMGB1) is a ubiquitous nuclear protein with multi-functions and plays an important role in tumorigenesis and metastasis in various human cancers. In the present study, we found that HMGB1 induced migration of in human non-small cell lung cancer (NSCLC) cells by up-regulating integrin alpha v beta 3 expression. Further investigation evidenced that HMGB1 activated Toll-like receptor 4 (TLR4) and NF-kappa B, which was responsible for alpha v beta 3 up-regulation. Furthermore, HMGB1-induced integrin alpha v beta 3 expression led to focal adhesion kinase (FAK) phosphorylation and increased paxillin and talin mRNA expression. Knockdown HMGB1 inhibited xenograft tumor metastasis in athymic mice. Taken together, our findings suggested that HMGB1 enhances tumor cell migration ability by activating alpha v beta 3/FAK through TLR4/NF-kappa B signaling, leading to metastasis of NSCLC. (C) 2016 Elsevier Inc. All rights reserved.