Expression of MUC1 mucin gene by hamster tracheal surface epithelial cells in primary culture

Abstract

Primary hamster tracheal surface epithelial (HTSE) cells carry mucin-like glycoproteins on the apical surface which are releasable by neutrophil elastase. In some cancer cells, mucins are localized on the cell surface and have been shown to be encoded by the MUC1 mucin gene. The objectives of the present experiments were: (I) to determine if HTSE cells express MUC1 mucin gene; (2) if they do, to isolate and characterize the hamster MUC1 complementary DNA (cDNA); and (3) to examine the pattern of MUC1 mRNA expression at different stages of culture. Reverse transcriptase-polymerase chain reaction amplification of HTSE cell RNAs using degenerate primers based on homologous sequences between the human and mouse MUC1 genes revealed the presence of a cDNA (0.5 kb) which has an 88% similarity in sequence with the mouse MUC1 cDNA. Using this 0.5 kb cDNA as a probe, an HTSE cell cDNA library was screened to isolate a hamster MUC1 cDNA clone. Sequence analysis of the cDNA revealed that it encodes an integral membrane protein of 676 amino acids which consists of (1) an N-terminal signal sequence, (2) the tandem repeat domain encoding 12 repeats of 20 amino acids, and (3) the C-terminal region consisting of degenerate tandem repeats and a unique sequence containing both the transmembrane and cytoplasmic domains. The presence of seven tyrosine residues in the cytoplasmic domain suggests a potential role as a receptor. Finally, expression of MUC1 mucin gene in HTSE cells appears to be associated with differentiation of secretory cells.