Glucocorticoids regulate surfactant protein synthesis in a pulmonary adenocarcinoma cell line

M. A. O'Reilly, A. F. Gazdar, J. C. Clark, T. J. Pilot-Matias, S. E. Wert, W. M. Hull, J. A. Whitsett

Research output: Contribution to journalArticle

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Abstract

Synthesis of pulmonary surfactant proteins SP-A, SP-B, and SP-C was demonstrated in a cell line derived from a human adenocarcinoma of the lung. The cells contained numerous lamellar inclusion bodies and formed organized groups of cells containing well-developed junctional complexes and apical microvillous membranes. Synthesis of SP-A was detected in the cells by enzyme-linked immunoabsorbent assay assay and by immunoprecipitation of [35S]methionine-labeled protein. SP-A was identified as an M(r) 31,000-36,000 polypeptide containing asparagine-linked carbohydrate. Northern blot analysis detected SP-A mRNA of 2.2 kb. Dexamethasone (1-10 nM) enhanced the relative abundance of SP-A mRNA. Despite stimulation of SP-A mRNA, intracellular SP-A content was unaltered or inhibited by dexamethasone. SP-B and SP-C mRNAs and synthesis of the SP-B and SP-C precursors were markedly induced by dexamethasone. ProSP-B was synthesized and secreted primarily as an M(r) 42,000-46,000 polypeptide. Proteolysis of the proSP-B resulted in the generation of endoglycosidase F-sensitive M(r) = 19,000-21,000 and 25,000-27,000 peptides, which were detected both intra- and extracellularly. SP-C proprotein of M(r) = 22,000 and smaller SP-C fragments were detected intracellularly but were not detected in the media. Mature forms of SP-B (M(r) = 8,000) and SP-C (M(r) = 4,000) were not detected. Glucocorticoids directly enhance the relative synthesis and mRNA of the surfactant proteins SP-A, SP-B, and SP-C. Discrepancies among SP-A mRNA, its de novo synthesis, and cell content suggest that glucocorticoid may alter both pre- and posttranslational factors modulating SP-A expression.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume257
Issue number6
StatePublished - 1989

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Surface-Active Agents
Glucocorticoids
Cell Line
Messenger RNA
Dexamethasone
Proteins
Peptides
Pulmonary Surfactant-Associated Proteins
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Pulmonary Surfactant-Associated Protein A
Asparagine
Inclusion Bodies
Immunoprecipitation
Northern Blotting
Methionine
Proteolysis
Adenocarcinoma of lung
Carbohydrates
Membranes
Enzymes

Keywords

  • enzyme-linked immunoabsorbent assay
  • Northern blot analysis

ASJC Scopus subject areas

  • Cell Biology
  • Physiology
  • Pulmonary and Respiratory Medicine

Cite this

O'Reilly, M. A., Gazdar, A. F., Clark, J. C., Pilot-Matias, T. J., Wert, S. E., Hull, W. M., & Whitsett, J. A. (1989). Glucocorticoids regulate surfactant protein synthesis in a pulmonary adenocarcinoma cell line. American Journal of Physiology - Lung Cellular and Molecular Physiology, 257(6).

Glucocorticoids regulate surfactant protein synthesis in a pulmonary adenocarcinoma cell line. / O'Reilly, M. A.; Gazdar, A. F.; Clark, J. C.; Pilot-Matias, T. J.; Wert, S. E.; Hull, W. M.; Whitsett, J. A.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 257, No. 6, 1989.

Research output: Contribution to journalArticle

O'Reilly, MA, Gazdar, AF, Clark, JC, Pilot-Matias, TJ, Wert, SE, Hull, WM & Whitsett, JA 1989, 'Glucocorticoids regulate surfactant protein synthesis in a pulmonary adenocarcinoma cell line', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 257, no. 6.
O'Reilly, M. A. ; Gazdar, A. F. ; Clark, J. C. ; Pilot-Matias, T. J. ; Wert, S. E. ; Hull, W. M. ; Whitsett, J. A. / Glucocorticoids regulate surfactant protein synthesis in a pulmonary adenocarcinoma cell line. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 1989 ; Vol. 257, No. 6.
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AU - Gazdar, A. F.

AU - Clark, J. C.

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AU - Wert, S. E.

AU - Hull, W. M.

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