The Role Of Receptor For Advanced Glycation End Products (RAGE) In The Biological Behavior Changes Of Human Hepatocellular Cancer Cell Line

Background:The receptor for advanced glycation end products (RAGE) is oncogenic and overexpressed in various human cancers, but its role in hepatocellular carcinoma remains unclear.Methods:We investigated RAGE expression in hepatocellular carcinoma Huh7and HepG2cell lines. The results indicated that Huh7cells express higher levels of RAGE mRNA than HepG2cells. Accordingly, we chose Huh7cells to investigate the role of RAGE in the proliferation of hepatocellular carcinoma. We transfected Huh7cells with RAGE-specific silencing RNA (siRNA), treated them with RAGE blocking antibody or treated them with high-mobility group box1protein (HMGB1).Results:We demonstrated that RAGE regulates cellular proliferation in HCC. Knockdown of RAGE by specific siRNA or hindering RAGE function by RAGE blocking antibody, inhibits cellular growth in the hepatocellular carcinoma cell line, Huh7, whereas the RAGE ligand, high mobility group box1protein (HMGB1) increases cellular proliferation. In addition, knockdown of RAGE by siRNA arrests cells in the G1phase and inhibits DNA synthesis (p<0.01), while HMGB1decreases the number of cells in the G1phase and increases the number in the S phase (p<0.05). Furthermore, RT-PCR, qRT-PCR and Western Blot results demonstrated that RAGE and HMGB1positively regulate nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB p65) expression in Huh7cells. Moreover, staining Huh7cells with Hoechst33258Stain demonstrated that RAGE antibody induces apoptosis, while HMGB1inhibits apoptosis. In addition, RT-PCR results indicated that RAGE antibody decreases the expression of cyclin D1and PCNA mRNA in Huh7cells.Conclusion:In this study we demonstrated that interfering with RAGE activity through the use of RAGE specific siRNA or RAGE blocking antibody could be of great benefit in reducing the proliferation of hepatocellular carcinoma. This suggests that RAGE and RAGE ligands are important targets for therapeutic intervention in hepatocellular carcinoma.