Adhesion of carcinoma cells to rat hepatocytes and rat fibronectin is inhibited by the OPAR monoclonal antibody, which is directed against a rat liver-specific carbohydrate epitope.
The OPAR mouse monoclonal antibody (mAb) directed against rat hepatocytes was previously shown to inhibit adhesion of TA3/Ha mammary carcinoma cells to hepatocytes. The antigen is abundantly present at the surface of hepatocytes beneath the endothelium of liver capillaries where we have observed invasion of carcinoma cells to occur. The OPAR mAb reacted with three major bands on a Western blot of liver plasma membrane proteins. The same proteins were also seen upon immunoprecipitation from iodinated liver plasma membrane proteins. We have isolated OPAR antigens by lectin wheat germ agglutinin (WGA) and OPAR affinity chromatography. Amino acid sequence analysis revealed that two of the bands were alpha 1-macroglobulin and C4-binding protein, which are serum components produced by hepatocytes. The presence of the epitope on distinct proteins and our previous observation that it can be detected in the Golgi apparatus but not in the endoplasmic reticulum, suggested that OPAR reacts with a liver-specific glycoconjugate. Loss of OPAR reactivity after neuraminidase and N-glycosidase F treatment showed that the epitope contains sialic acid residues on N-linked sugar moieties. OPAR also reacted with rat fibronectin, and inhibited adhesion of TA3/St cells to fibronectin. This explains the inhibition by the OPAR mAb of TA3/St cell adhesion to hepatocytes, which we have shown to be due mainly to interaction with hepatocyte surface-associated fibronectin. However, adhesion of the related TA3/Ha cells to hepatocytes, which is mediated by the alpha 6 beta 4 integrin, and does not involve binding to fibronectin, is also inhibited. This suggests that alpha 6 beta 4 on liver-metastasizing carcinoma cells binds to an OPAR epitope-carrying glycoprotein produced by hepatocytes.
Kemperman
H
H
Division of Cell Biology, The Netherlands Cancer Institute, Amsterdam.
Wijnands
Y
Y
Roos
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology*; D002448:Cell Adhesion / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology; D000939:Epitopes / Q000032:analysis; D005353:Fibronectins / Q000276:immunology; D006026:Glycoside Hydrolases; D008099:Liver / Q000166:cytology / Q000276:immunology*; D008325:Mammary Neoplasms, Experimental / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009439:Neuraminidase; D051381:Rats; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D000939:Epitopes; D005353:Fibronectins; D006026:Glycoside Hydrolases; D009439:Neuraminidase