Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells.
<h4>Background</h4>Expression of surfactant protein (SP)-B, which assures the structural stability of the pulmonary surfactant film, is influenced by various stimuli, including glucocorticoids; however, the role that cell-cell contact plays in SP-B transcription remains unknown. The aim...
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Public Library of Science (PLoS)
2017-01-01
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| author | Markus Fehrholz Silvia Seidenspinner Steffen Kunzmann |
| author_facet | Markus Fehrholz Silvia Seidenspinner Steffen Kunzmann |
| author_sort | Markus Fehrholz |
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| description | <h4>Background</h4>Expression of surfactant protein (SP)-B, which assures the structural stability of the pulmonary surfactant film, is influenced by various stimuli, including glucocorticoids; however, the role that cell-cell contact plays in SP-B transcription remains unknown. The aim of the current study was to investigate the impact of cell-cell contact on SP-B mRNA and mature SP-B expression in the lung epithelial cell line H441.<h4>Methods</h4>Different quantities of H441 cells per growth area were either left untreated or incubated with dexamethasone. The expression of SP-B, SP-B transcription factors, and tight junction proteins were determined by qPCR and immunoblotting. The influence of cell density on SP-B mRNA stability was investigated using the transcription inhibitor actinomycin D.<h4>Results</h4>SP-B mRNA and mature SP-B expression levels were significantly elevated in untreated and dexamethasone-treated H441 cells with increasing cell density. High cell density as a sole stimulus was found to barely have an impact on SP-B transcription factor and tight junction mRNA levels, while its stimulatory ability on SP-B mRNA expression could be mimicked using SP-B-negative cells. SP-B mRNA stability was significantly increased in high-density cells, but not by dexamethasone alone.<h4>Conclusion</h4>SP-B expression in H441 cells is dependent on cell-cell contact, which increases mRNA stability and thereby potentiates the glucocorticoid-mediated induction of transcription. Loss of cell integrity might contribute to reduced SP-B secretion in damaged lung cells via downregulation of SP-B transcription. Cell density-mediated effects should thus receive greater attention in future cell culture-based research. |
| format | Article |
| id | doaj-art-94afc0eea7e14136ab079f5cee4affc7 |
| institution | DOAJ |
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| language | English |
| publishDate | 2017-01-01 |
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| spelling | doaj-art-94afc0eea7e14136ab079f5cee4affc72025-08-20T02:46:01ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01129e018455610.1371/journal.pone.0184556Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells.Markus FehrholzSilvia SeidenspinnerSteffen Kunzmann<h4>Background</h4>Expression of surfactant protein (SP)-B, which assures the structural stability of the pulmonary surfactant film, is influenced by various stimuli, including glucocorticoids; however, the role that cell-cell contact plays in SP-B transcription remains unknown. The aim of the current study was to investigate the impact of cell-cell contact on SP-B mRNA and mature SP-B expression in the lung epithelial cell line H441.<h4>Methods</h4>Different quantities of H441 cells per growth area were either left untreated or incubated with dexamethasone. The expression of SP-B, SP-B transcription factors, and tight junction proteins were determined by qPCR and immunoblotting. The influence of cell density on SP-B mRNA stability was investigated using the transcription inhibitor actinomycin D.<h4>Results</h4>SP-B mRNA and mature SP-B expression levels were significantly elevated in untreated and dexamethasone-treated H441 cells with increasing cell density. High cell density as a sole stimulus was found to barely have an impact on SP-B transcription factor and tight junction mRNA levels, while its stimulatory ability on SP-B mRNA expression could be mimicked using SP-B-negative cells. SP-B mRNA stability was significantly increased in high-density cells, but not by dexamethasone alone.<h4>Conclusion</h4>SP-B expression in H441 cells is dependent on cell-cell contact, which increases mRNA stability and thereby potentiates the glucocorticoid-mediated induction of transcription. Loss of cell integrity might contribute to reduced SP-B secretion in damaged lung cells via downregulation of SP-B transcription. Cell density-mediated effects should thus receive greater attention in future cell culture-based research.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184556&type=printable |
| spellingShingle | Markus Fehrholz Silvia Seidenspinner Steffen Kunzmann Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells. PLoS ONE |
| title | Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells. |
| title_full | Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells. |
| title_fullStr | Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells. |
| title_full_unstemmed | Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells. |
| title_short | Expression of surfactant protein B is dependent on cell density in H441 lung epithelial cells. |
| title_sort | expression of surfactant protein b is dependent on cell density in h441 lung epithelial cells |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0184556&type=printable |
| work_keys_str_mv | AT markusfehrholz expressionofsurfactantproteinbisdependentoncelldensityinh441lungepithelialcells AT silviaseidenspinner expressionofsurfactantproteinbisdependentoncelldensityinh441lungepithelialcells AT steffenkunzmann expressionofsurfactantproteinbisdependentoncelldensityinh441lungepithelialcells |