Evidence of mucin secretion in human lung adenocarcinoma cell lines NCIH650 and NCIH2077 and effect of select secretagogues on mucin secretion

Prerna M. Sharma, Mangala G. Sarkar, Arvind K. Virmani, Adi F. Gazdar, Goverdhan P. Sachdev

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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/actors 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 Superase 6 and Superase 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.

Original languageEnglish (US)
Pages (from-to)473-483
Number of pages11
JournalBioscience Reports
Volume19
Issue number5
StatePublished - 1999

Fingerprint

Mucins
Cells
Cell Line
Molecular Weight
Molecular weight
Leukocyte Elastase
Glycoconjugates
Adenosine Monophosphate
Cell culture
Adenocarcinoma of lung
Mucin-1
Ionomycin
Glucosamine
Ammonium Sulfate
Gene Expression Regulation
Neoplasm Antigens
Tretinoin
Chromatography
Gene expression
Culture Media

Keywords

  • Gene expression
  • Lung adenocarcinoma
  • Lung cancer
  • Mucin
  • Secretion

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Cell Biology

Cite this

Evidence of mucin secretion in human lung adenocarcinoma cell lines NCIH650 and NCIH2077 and effect of select secretagogues on mucin secretion. / Sharma, Prerna M.; Sarkar, Mangala G.; Virmani, Arvind K.; Gazdar, Adi F.; Sachdev, Goverdhan P.

In: Bioscience Reports, Vol. 19, No. 5, 1999, p. 473-483.

Research output: Contribution to journalArticle

Sharma, Prerna M. ; Sarkar, Mangala G. ; Virmani, Arvind K. ; Gazdar, Adi F. ; Sachdev, Goverdhan P. / Evidence of mucin secretion in human lung adenocarcinoma cell lines NCIH650 and NCIH2077 and effect of select secretagogues on mucin secretion. In: Bioscience Reports. 1999 ; Vol. 19, No. 5. pp. 473-483.
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