Role of galaptin in ovarian carcinoma adhesion to extracellular matrix in vitro.
Immunohistochemical studies indicated that galaptin is a major protein of ovarian carcinoma cells present in patient effusions and it is distributed throughout the cytoplasm. Enzyme-linked immunoadsorbent assay (ELISA) and immunoprecipitation experiments demonstrated that galaptin is also a major protein of the A121 ovarian carcinoma cell line, constituting less than or equal to 1% of extractable protein bound by DEAE Sephacel. Western blot analyses revealed that the galaptin present in ovarian carcinoma consists of a 14.5 KD subunit. Ovarian carcinoma and mesothelial cells isolated from patient effusions display surface receptors for galaptin with an apparently greater density of receptors present on the carcinoma cells. A121 cells also display surface receptors for galaptin: binding sites/cell = 3 X 10(8) and Ka = 1.2 X 10(9) M-1. The presence of galaptin in bovine corneal endothelial cells (BCEC) and BCEC-derived extracellular matrix (ECM) was demonstrated by ELISA. Of the total ECM-bound galaptin, about 75% appears to be insoluble in phosphate-buffered saline (PBS) lactose. ECM was also found to contain abudnant receptors for galaptin. Treatment of ECM with lactose increased the apparent galaptin receptor density:binding sites/cm2 = 7 X 10(13) and Ka = 2.6 X 10(9) M-1. Pretreatment of A121 cells with galaptin inhibited adhesion to ECM. The addition of exogenous galaptin to ECM had variable effect on cell adhesion. The data presented here suggest that early adhesion events may be carbohydrate-specific involving interaction between ECM-bound galaptin and cell surface galaptin receptors.
Department of Surgical Oncology, Roswell Park Memorial Institute, Buffalo, New York 14263.
H J
HJ
Allen
D
D
Sucato
B
B
Woynarowska
S
S
Gottstine
A
A
Sharma
R J
RJ
Bernacki
D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002417:Cattle; D002448:Cell Adhesion; D004728:Endothelium, Corneal; D004797:Enzyme-Linked Immunosorbent Assay; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D037161:Galectins; D006388:Hemagglutinins / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D007106:Immune Sera; D007124:Immunoenzyme Techniques; D010051:Ovarian Neoplasms / Q000378:metabolism*; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured
D037161:Galectins; D006388:Hemagglutinins; D007106:Immune Sera