V domain of RAGE interacts with AGEs on prostate carcinoma cells.
BACKGROUND
The expression of the scavenger receptor for advanced glycation end products (RAGE) and various ligands of RAGE correlate significantly with cancer progression. However, the mechanism of RAGE/sRAGE-induced cancer cell activation and ligand usage remain largely unknown.


METHODS
Androgen-independent, highly invasive, as well as androgen-dependent, non-invasive human prostate carcinoma (CaP) cells were investigated for their interaction with the soluble form of RAGE (sRAGE). Using neutralizing antibodies and soluble proteins, the ligand for RAGE was identified on CaP cells and ligand binding with sRAGE was biochemically characterized.


RESULTS
Both androgen-independent, highly invasive and androgen-dependent, non-invasive CaP cells interacted with immobilized sRAGE in a surprisingly strong manner. Using C-terminal truncation variants of RAGE we identified the V domain being responsible for the adhesion of CaP cells to sRAGE. Moreover, we demonstrate that this adhesion cannot be blocked by S100B or neutralizing antibodies against beta integrins, or amphoterin. However, the CaP cell-RAGE interaction was inhibited with either AGE-modified proteins, or with neutralizing antibodies against AGE or RAGE. Despite similar binding kinetics between AGE-modified BSA and different RAGE domains, only applying an excess of sRAGE, but not the VC1 or V domain of RAGE, was able to block the CaP cell-RAGE interaction.


CONCLUSIONS
We identified AGEs as the ligand for RAGE on both invasive and non-invasive prostate cancer cells.
Allmen
Edith Uetz-von
EU
Biotechnology Institute Thurgau, University of Konstanz, Kreuzlingen, Switzerland.
Koch
Michael
M
Fritz
Günter
G
Legler
Daniel F
DF
D019138:Antibodies, Blocking / Q000494:pharmacology; D001665:Binding Sites; D001667:Binding, Competitive; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D017127:Glycation End Products, Advanced / Q000737:chemistry / Q000378:metabolism*; D024243:HMGB1 Protein / Q000276:immunology; D006801:Humans; D008024:Ligands; D008297:Male; D009414:Nerve Growth Factors / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000276:immunology / Q000378:metabolism*; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000378:metabolism
D019138:Antibodies, Blocking; D017127:Glycation End Products, Advanced; D024243:HMGB1 Protein; D008024:Ligands; D009414:Nerve Growth Factors; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; C579160:S100B protein, human