S100A9 promotes human hepatocellular carcinoma cell growth and invasion through RAGE-mediated ERK1/2 and p38 MAPK pathways.

S100A9 belongs to the S100 family of calcium-binding proteins and is over-expressed in many human tumors including hepatocellular carcinoma (HCC). Recent study demonstrated that S100A9 is significantly elevated and is associated with tumor differentiation and vascular invasion in HCC. The functional role of S100A9 is, however, poorly understood. Here, we demonstrated that S100A9 treatment increased viability, invasiveness and clone formation in three HCC cell lines (HepG2, SMMC-7721 and Huh7). S100A9 also promoted tumor growth in vivo by a xenograft mouse model. In addition, we observed a co-localization of S100A9 with receptor for advanced glycation end products (RAGE) in human HCC intratumoral tissues and an interaction of S100A9 with RAGE in vitro. Treatment with RAGE blocking antibody blocked the enhanced viability, invasion, clone formation and tumor growth in vivo resulted by S100A9, suggesting that these effects were mediated via RAGE ligation. In order to investigate the signaling pathways, mitogen-activated protein kinase (MAPK) phosphorylation was characterized. S100A9 caused a significant increase in p-p38 and p-ERK1/2 levels, and inhibition of which blocked enhanced invasion and viability resulted by S100A9, respectively. Furthermore, treatment with RAGE blocking antibodies also abrogated the S100A9-induced p38 and ERK1/2 activation, suggesting that S100A9-induced MAPK activation is mediated via RAGE ligation. Our data demonstrate that S100A9 binds to RAGE and stimulates RAGE-dependent MAPK signaling cascades, promoting cell growth and invasion in HCC.