Inhibitory Effects Of Blockage Targeting At RAGE-ligand Axis On Invasion And Proliferation Of Pancreatic Neoplasms Cell Line PANC-1

BACKGROUND & OBJECTIVE: Receptor for advanced glycation end-products (RAGE) is a multifunctional receptor with a wide array of lignads that seem to dictate its involvement in several diseases, including diabetes, inflammation, and Alzheimer's disease. Recent evidence indicates that RAGE and its ligands also have an important role in cancer. RAGE-ligand axis can interact in an autocrine manner to directly activate cancer cells and stimulate proliferation, invasion, metastasis, and chemoresistance. This study is to investigate RAGE expression in high-invasive pancreatic neoplasms cell line PANC-1, evaluate the inhibitory effects of RAGE-ligand biological axis blockage via Monoclonal antibody method on the proliferation and invasion activity of PANC-1 in vitro, and explore the related molecular mechanism.Methods:①RAGE mRNA and protein expression in PANC-1 cell was seperatively detected by RT-PCR and flow cytometery (FCM).②Anti-RAGE McAb with different concentrations were cultured with PANC-1 cell in vitro. Cell growth change was examined by MTT. Flow cytometery (FCM) shew the cell cycle and apoptosis rate change. Apoptosis-associated protein such as NF-κb, Bcl-2 and Caspase-3 expression were assessed through cell immunohistochemistry(IHC).③Cell invasion ability was evaluated by Transwell Invasion Assay. Semi-quantitative RT-PCR,ELISA Assay and IHC observed the change of MMP-2,9 expression in gene and protein level.Results:①RAGE was obviously expressed in the surface of PANC-1 cell.②Anti-RAGE McAb inhibited the cell proliferation and induce cell apoptosis, mainly retarded in G1 cell cycle, this effect was increased with a dose and time dependent manner. Anti-RAGE McAb increased the ratio in G1 stage of cell cycle, apoptosis rate and NF-κb,Bcl-2 expression were down-regulated highly, on the other hand ,Caspase-3 was activated subsequencely.③Anti-RAGE McAb inhibited the cell invasion ability, this effect was increased with a dose dependent manner. At the same time, MMP-2,9 expression were effectively suppressed.Conclusions: The expression of RAGE is strongly positive in the surface of PANC-1 cell. With the RAGE-ligand system, Anti-RAGE McAb blocked this RAGE-mediated signal pathway, hindered PANC-1 cell growth and invasion through down-regulation of NF-κb,Bcl-2,MMP-2,9 and up-regulation of Caspase-3. All these observations suggest RAGE-ligand axis can interact in an autocrine manner to sustain the autonomous growth of PANC-1 cell, and anti-RAGE McAb could partially intervene these biological functions.