Expression of fetoacinar pancreatic (FAP) protein in the pancreatic human tumor cell line BxPC-3.
Fetoacinar pancreatic (FAP) protein is a specific component of the human exocrine pancreas that may have a role in the differentiation and transformation of this organ. In order to set out a model for studies on the regulation of FAP, 47 established cell lines from human cancer of different origins were tested for FAP expression using the monoclonal antibody J28 (Mab J28). Only two, both pancreatic, were positive. This finding supports the already reported pancreatic specificity of this antigen. Strongest expression was shown by the BxPC-3 cell line, derived from a moderately well-differentiated adenocarcinoma in the body of the pancreas. In BxPC-3 cells grown in Roswell Park Memorial Institute (RPMI) 1640-10% fetal bovine serum (FBS), Mab J28 immunostaining was localized in the cytoplasm of the cells. In serum-free medium, cells quickly died. Growth and FAP expression were maintained when this medium was supplemented with insulin. FAP is not released to the culture medium, as evidence by absence of reaction with the monoclonal antibody on nitrocellulose dot-blots. On the contrary, a positive reaction was observed in cell homogenates made by sonication or by extraction with 0.1% Triton. A competitive enzyme-linked immunosorbent assay (ELISA), using biotinylated FAP, was developed to quantify the protein in cell homogenates. Concentrations of FAP in homogenates from cells cultured in standard conditions or serum-free supplemented with insulin were in the range of 0.28-0.40 micrograms FAP/mg total protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Mazo
A
A
Departamento de Bioquímica i Fisiología, Facultad de Química, Universidad de Barcelona, Spain.
Fujii
Y
Y
Shimotake
J
J
Escribano
M J
MJ
D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D001710:Biotin; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism*; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D008049:Lipase; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D003470:Culture Media; D006023:Glycoproteins; D007328:Insulin; D001710:Biotin; C054621:CEL protein, human; D008049:Lipase