Evidence of mucin secretion in human lung adenocarcinoma cell lines NCIH650 and NCIH2077 and effect of select secretagogues on mucin secretion.
Mucins comprise an important class of tumor-associated antigens. The objectives of the present study were (a) to establish an in vitro model system using human non-small cell lung adenocarcinoma cell lines NCIH650 and NCIH2077 (b) provide evidence that these cell lines secrete mucin in culture conditions and (c) investigate the effects of select secretagogues on mucin secretion. The cell lines were established in ACL-4 medium containing several growth factors and retinoic acid and 5% fetal calf serum. The high molecular weight glycoconjugates secreted in the culture medium were purified by ammonium sulfate precipitation and Superose 6 and Superose 12 FPLC chromatography. The purified high molecular weight glycoconjugate fraction and the carcinoma cells were shown to have mucin by dot blot, Western blot and immunohistochemical analysis, respectively, using specific antibodies to purified major mucin, HTM-1. Also, incorporation experiments with mucin precursor 3H-glucosamine demonstrated that the cells indeed synthesize high molecular weight mucins. The effects of secretagogues such as, 8-bromocyclic AMP, ionomycin, phorbol-12-myristate-13-acetate and neutrophil elastase on mucin secretion were also investigated. Only 8-bromocyclic AMP and neutrophil elastase influenced mucin secretion. These studies provided strong evidence that the lung adenocarcinoma cell lines secrete high molecular weight mucins in culture conditions and only two of the four tested secretagogues significantly increased mucin secretion. Thus, this in vitro model system may be useful in determining alterations in mucin structure, if any, in lung adenocarcinomas as well as in studying the regulation of mucin gene expression.
Sharma
P M
PM
College of Pharmacy, University of Oklahoma Health Sciences Center, Oklahoma City 73190, USA.
Sarkar
M G
MG
Virmani
A K
AK
Gazdar
A F
AF
Sachdev
G P
GP
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000230:Adenocarcinoma; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005944:Glucosamine / Q000378:metabolism; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology; D019272:Leukocyte Elastase / Q000494:pharmacology; D008175:Lung Neoplasms; D009077:Mucins / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014316:Tritium; D014407:Tumor Cells, Cultured
D009077:Mucins; D014316:Tritium; D015124:8-Bromo Cyclic Adenosine Monophosphate; D015759:Ionomycin; D019272:Leukocyte Elastase; D005944:Glucosamine; D013755:Tetradecanoylphorbol Acetate