TY - JOUR
T1 - Differential effects of glucocorticoid on expression of surfactant proteins in a human lung adenocarcinoma cell line
AU - O'Reilly, Michael A.
AU - Gazdar, Adi F.
AU - Morris, Randal E.
AU - Whitsett, Jeffrey A.
N1 - Funding Information:
This work was supported by HL 28623 and HL 38859. M.A.O'R. is funded in part by a Molecular and Cellular Biology Training Grant HL 07527.
PY - 1988/6/30
Y1 - 1988/6/30
N2 - Synthesis of pulmonary surfactant-associated glycoproteins of Mr 28 000-36 000 (SP-A) and Mr 42 000-46 000 (proSP-B) has been identified in a continuous cell line derived from a human lung adenocarcinoma. SP-A was detected by immunoblot analysis, ELISA assay and by [35S]methionine labelling of the cells. SP-A was secreted into the media as an endoglycosidase F sensitive glycoprotein which co-migrated with the isoforms of SP-A identified in human lavage fluid by 2D-IEF-SDS-PAGE. Hybridization of cellular RNA with SP-A-specific cDNA identified an abundant 2.2 kb mRNA species, identical to that observed in human lung. SP-A RNA and protein content were markely inhibited by dexamethasone in a dose-dependent fashion. Under identical culture conditions, synthesis of a distinct surfactant protein, SP-B, was markedly stimulated by the glucocorticoid. The SP-B precursor was secreted into the media as heterogenous Mr 42 000-46 00 protein, pI 4.6-5.1, and was sensitive to endoglycosidase F. Synthesis of proSP-B was enhanced by the glucocorticoid in a dose-dependent fashion and was associated with increased SP-B mRNA of 2.0 kb detected by Northern blot analysis. The cell line secreted proSP-B as Mr 42 000-46 000 glycosylated protein and did not process the precursor to the Mr 7000-8000 and secretes peptide. In summary, a human adenocarcinoma cell line has been identified which synthesized and secretes two surfactant-associated proteins, SP-A and proSP-B. Glucocorticoid enhanced SP-B but inhibited SP-A expression in this cell line. The identification of a continuous cell line secreting surfactant proteins may be useful in the study of synthesis and secretion of these important proteins and for production of the proteins for clinical uses.
AB - Synthesis of pulmonary surfactant-associated glycoproteins of Mr 28 000-36 000 (SP-A) and Mr 42 000-46 000 (proSP-B) has been identified in a continuous cell line derived from a human lung adenocarcinoma. SP-A was detected by immunoblot analysis, ELISA assay and by [35S]methionine labelling of the cells. SP-A was secreted into the media as an endoglycosidase F sensitive glycoprotein which co-migrated with the isoforms of SP-A identified in human lavage fluid by 2D-IEF-SDS-PAGE. Hybridization of cellular RNA with SP-A-specific cDNA identified an abundant 2.2 kb mRNA species, identical to that observed in human lung. SP-A RNA and protein content were markely inhibited by dexamethasone in a dose-dependent fashion. Under identical culture conditions, synthesis of a distinct surfactant protein, SP-B, was markedly stimulated by the glucocorticoid. The SP-B precursor was secreted into the media as heterogenous Mr 42 000-46 00 protein, pI 4.6-5.1, and was sensitive to endoglycosidase F. Synthesis of proSP-B was enhanced by the glucocorticoid in a dose-dependent fashion and was associated with increased SP-B mRNA of 2.0 kb detected by Northern blot analysis. The cell line secreted proSP-B as Mr 42 000-46 000 glycosylated protein and did not process the precursor to the Mr 7000-8000 and secretes peptide. In summary, a human adenocarcinoma cell line has been identified which synthesized and secretes two surfactant-associated proteins, SP-A and proSP-B. Glucocorticoid enhanced SP-B but inhibited SP-A expression in this cell line. The identification of a continuous cell line secreting surfactant proteins may be useful in the study of synthesis and secretion of these important proteins and for production of the proteins for clinical uses.
KW - (Human lung)
KW - Corticosteroid
KW - Gene expression
KW - Pulmonary surfactant
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U2 - 10.1016/0167-4889(88)90179-6
DO - 10.1016/0167-4889(88)90179-6
M3 - Article
C2 - 3382698
AN - SCOPUS:0023938083
SN - 0167-4889
VL - 970
SP - 194
EP - 204
JO - BBA - Molecular Cell Research
JF - BBA - Molecular Cell Research
IS - 2
ER -