7545050

pubmed:Citation

2

The purpose of this study was to determine the biochemical and molecular characteristics of mucin synthesized by cystic fibrosis cells (CFPAC-1), a pancreatic cancer cell line derived from a patient with cystic fibrosis, and pancreatic cancer (SW-1990) cell lines. High molecular weight glycoproteins (HMG) were quantified by [3H]-glucosamine labeling and chromatography on sepharose CL-4B. Mucin gene expression was determined by using cDNA probes for 2 distinct intestinal mucins (MUC2 and MUC3) and one stomach mucin (MUC1). The specific mucin core epitopes were confirmed by immunoblots using antibodies that recognize T, Tn, sialosyl Tn, MUC1, MUC2, and MUC3. The results of these experiments demonstrate that CFPAC-1 cells contained 1.25 fold and 1.4 fold more HMG in the membrane and cytosolic fractions, however, secreted 4-fold more HMG into the medium compared to SW-1990 cells. The HMG of SW-1990 was found to be mucinous in nature and not proteoglycans, as it was not susceptible to hyalurinidase, heparinase and chondroitinase ABC. The HMG of CFPAC-1 was also predominantly (80%) mucinous but with small amounts of proteoglycans. mRNA and immunoblot analysis suggest that these CFPAC-1 and SW-1990 cells predominantly express MUC1 apomucin, small amounts of MUC2 apomucin, and no MUC3. Pulse chase labeling and immunoprecipitation of MUC1 type mucin using the 139H2 monoclonal antibody demonstrated that different sizes of mucin gene product were present in both cell lines, corresponding to the known length polymorphism of this mucin. Both T and Tn antigens were significantly higher in CFPAC-1 and SW-1990 cells as compared to sialosyl Tn antigen. These findings were associated with the increased activities of polypeptidyl N-acetylgalactosaminyl transferase and b1,3-galactosyltransferase. These investigations demonstrate for the first time that cystic fibrosis cells (CFPAC-1) secrete and synthesize high amounts of mucin which is associated with high levels of MUC1 mRNA, low levels of MUC2 mRNA and non detectable MUC3 mRNA.

eng

IM

MEDLINE

1039-9712

Print

NLM

351-62

pubmed-meshheading:7545050-Amino Acid Sequence, pubmed-meshheading:7545050-Antibodies, pubmed-meshheading:7545050-Cell Line, pubmed-meshheading:7545050-Chondroitinases and Chondroitin Lyases, pubmed-meshheading:7545050-Consensus Sequence, pubmed-meshheading:7545050-Cystic Fibrosis, pubmed-meshheading:7545050-DNA, Complementary, pubmed-meshheading:7545050-DNA Probes, pubmed-meshheading:7545050-Epitopes, pubmed-meshheading:7545050-Gene Expression, pubmed-meshheading:7545050-Glucosamine, pubmed-meshheading:7545050-Glycoproteins, pubmed-meshheading:7545050-Heparin Lyase, pubmed-meshheading:7545050-Humans, pubmed-meshheading:7545050-Hyaluronoglucosaminidase, pubmed-meshheading:7545050-Immunoblotting, pubmed-meshheading:7545050-Membrane Glycoproteins, pubmed-meshheading:7545050-Molecular Sequence Data, pubmed-meshheading:7545050-Mucin-1, pubmed-meshheading:7545050-Mucins, pubmed-meshheading:7545050-Neoplasm Proteins, pubmed-meshheading:7545050-Pancreatic Neoplasms, pubmed-meshheading:7545050-Polysaccharide-Lyases, pubmed-meshheading:7545050-RNA, Messenger, pubmed-meshheading:7545050-Repetitive Sequences, Nucleic Acid, pubmed-meshheading:7545050-Tritium

Cystic fibrosis and pancreatic cancer cells synthesize and secrete MUC1 type mucin gene product.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S.