A cell-surface proteoglycan mediates human adenocarcinoma HT-29 cell adhesion to human angiogenin.
Human angiogenin is an excellent substrate for the adhesion of HT-29 human colon adenocarcinoma cells. These cells adhere more quickly to human angiogenin than to fibronectin, laminin, collagen I, and collagen IV. Anti-angiogenin antibodies and the angiogenesis inhibitors platelet factor-4 and placental ribonuclease inhibitor prevent adhesion of HT-29 cells to angiogenin. Calcium and magnesium ions are not required for adhesion and Arg-Gly-Asp-Ser has no effect, indicating that the interaction is integrin-independent. Instead, adhesion seems to involve a heparan/chondroitin sulfate proteoglycan. Treatment of the cells with heparinase or heparitinase decreases HT-29 cell adhesion onto angiogenin but not onto collagen I. Moreover, cell adhesion is decreased by the presence of heparin or chondroitin sulfates and by preincubation of the cells with inhibitors of proteoglycan synthesis or secretion. In addition, angiogenin binds tightly to heparin-Sepharose, requiring 0.78 M NaCl for elution. Angiogenin-affinity chromatography of a 35S-, 3H-labeled HT-29 cell fraction enriched in cell-surface proteoglycans yields a single, heparinase-sensitive component of apparent molecular mass > 200 kDa, as detected by autoradiography after SDS-polyacrylamide gel electrophoresis. These results suggest that angiogenin could be an effective substrate for tumor cell adhesion during metastasis and may provide a basis for the design of inhibitors of this process.
Soncin
F
F
Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Boston, Massachusetts 02115.
Shapiro
R
R
Fett
J W
JW
D000230:Adenocarcinoma / Q000737:chemistry / Q000473:pathology*; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000378:metabolism; D003110:Colonic Neoplasms / Q000737:chemistry / Q000473:pathology*; D006025:Glycosaminoglycans / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D008274:Magnesium / Q000378:metabolism; D009389:Neovascularization, Pathologic; D009842:Oligopeptides / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic; D014407:Tumor Cells, Cultured
D015815:Cell Adhesion Molecules; D006025:Glycosaminoglycans; D009842:Oligopeptides; D011506:Proteins; D011509:Proteoglycans; C046617:arginyl-glycyl-aspartyl-serine; C046366:angiogenin; D012259:Ribonuclease, Pancreatic; D008274:Magnesium; D002118:Calcium