EXPRESSION OF CCKB GASTRIN RECEPTOR ISOFORMS IN GASTROINTESTINAL TUMOR-CELLS/

Anti-serum raised against the human cholecystokinin B (CCKB)/gastrin r eceptor was used in Western blotting to differentiate the cellular loc ations of receptor isoforms expressed by human gastro-intestinal (GI) tumour cell lines. Using anti-serum directed against the amino-termina l extracellular tail of the CCKB/gastrin receptor, 8/9 cell lines were shown to express immunoreactive proteins of either m.w. 70 or 40 kDa, or both. Both isoforms were found to be associated with intracellular , non-nuclear membranes, whereas only the 70 kDa protein was expressed in the plasma membrane. Receptor expression was related to gastrin pr oduction and secretion. Both progastrin and glycine-extended gastrin-1 7 were produced and secreted by the tumour cell lines; however, carbox y amidated gastrin was not detected by radioimmunoassay. A CCKB/gastri n receptor transfectant NIH3T3 cell line did not produce detectable ga strin and showed exclusive expression of the 70 kDa receptor on the pl asma membrane. One cell line had <50 pg/ml cell-associated progastrin and no detectable receptor form. Cell lines expressing 50-150 pg/ml ha d both 40 and 70 kDa receptor forms. Those expressing > 150 pg/ml prog astrin had only the 40 kDa isoform, which was shown to be exclusively expressed on intracellular, non-nuclear membranes, in one of the cell lines. Of the 4 cell lines exclusively expressing the lower m.w. recep tor, 3 had gastrin present within the cell, which was not secreted. Th us, if cell- associated gastrin induces a proliferative effect, it may be by an intracrine pathway. Our study has identified the presence of CCKB/gastrin receptor isoforms in different cellular locations and ma y help toward understanding the complex autocrine and intracrine pathw ays mediated by gastrin peptides. Int. J. Cancer 77:572-577, 1998. (C) 1998 Wiley-Liss.