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Research article
TGF-β-Induced α-SMA Expression is Mediated by C/EBPβ Acetylation in Human Alveolar Epithelial Cells In Vitro
Zili Yi
Hunan Agricultural University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0123-7611
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This study sought to determine whether binding of acetylated C/EBPβ to α-SMA promoter could affect its activity and was essential for EMT and extracellular matrix deposition in IPF using in vitro model. The expression of EMT and C/EBPβ in A549 cells with TGF-β as pulmonary fibrotic model were detected by western blotting and qPCR. Collagen-I expression using ELISA was performed. The luciferase activity was used to examine the activity of C/EBPβ. Knockdown of C/EBPβ was performed by siRNA. We also investigated the effect of deacetylation of C/EBPβ on EMT using SIRT1. The binding ability of C/EBPβ with α-SMA promoter was affirmed via ChIP and EMSA. The relationship of α-SMA and acetylated C/EBPβ was investigated by Co-IP. SiRNA-mediated knockdown of C/EBPβ in A549 cells attenuated TGF-β1-induced myofibroblast differentiation and ECM deposition. The extent of association between acetylated C/EBPβ and α-SMA promoter was dynamically monitored. It was confirmed that deacetylation of C/EBPβ in A549 cells successfully ameliorated TGF-β1-induced EMT, as shown by reduction in α-SMA expression and excessive collagen-I accumulation. The EMT and fibrotic effect of TGF-β1 dependent on acetylated C/EBPβ-mediated regulation of α-SMA gene activity. C/EBPβ acetylation may play a central role in pulmonary fibrosis.
Keywords
C/EBPβ, Idiopathic Pulmonary Fibrosis, Collagen, α-SMA
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CURRENT STATUS: Under Review
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Posted 17 Sep, 2020
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Editorial decision: Major revision
On 06 Oct, 2020
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Received 03 Oct, 2020
Review #1 received
Received 28 Sep, 2020
Reviewer #2 agreed
On 18 Sep, 2020
Editor assigned
On 17 Sep, 2020
Reviewers invited
Invitations sent on 17 Sep, 2020
Reviewer #1 agreed
On 17 Sep, 2020
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On 16 Sep, 2020
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On 15 Sep, 2020
First submitted
On 11 Sep, 2020
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Subject Areas
Molecular Genetics
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This preprint is under consideration at Molecular Medicine. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
TGF-β-Induced α-SMA Expression is Mediated by C/EBPβ Acetylation in Human Alveolar Epithelial Cells In Vitro
Zili Yi
Hunan Agricultural University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0123-7611
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 17 Sep, 2020
No community comments so far
Editorial decision: Major revision
On 06 Oct, 2020
Review #2 received
Received 03 Oct, 2020
Review #1 received
Received 28 Sep, 2020
Reviewer #2 agreed
On 18 Sep, 2020
Editor assigned
On 17 Sep, 2020
Reviewers invited
Invitations sent on 17 Sep, 2020
Reviewer #1 agreed
On 17 Sep, 2020
Editor invited
On 16 Sep, 2020
Submission checks complete
On 15 Sep, 2020
First submitted
On 11 Sep, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Molecular Genetics
License:
This work is licensed under a CC BY 4.0 License. Read Full License
This study sought to determine whether binding of acetylated C/EBPβ to α-SMA promoter could affect its activity and was essential for EMT and extracellular matrix deposition in IPF using in vitro model. The expression of EMT and C/EBPβ in A549 cells with TGF-β as pulmonary fibrotic model were detected by western blotting and qPCR. Collagen-I expression using ELISA was performed. The luciferase activity was used to examine the activity of C/EBPβ. Knockdown of C/EBPβ was performed by siRNA. We also investigated the effect of deacetylation of C/EBPβ on EMT using SIRT1. The binding ability of C/EBPβ with α-SMA promoter was affirmed via ChIP and EMSA. The relationship of α-SMA and acetylated C/EBPβ was investigated by Co-IP. SiRNA-mediated knockdown of C/EBPβ in A549 cells attenuated TGF-β1-induced myofibroblast differentiation and ECM deposition. The extent of association between acetylated C/EBPβ and α-SMA promoter was dynamically monitored. It was confirmed that deacetylation of C/EBPβ in A549 cells successfully ameliorated TGF-β1-induced EMT, as shown by reduction in α-SMA expression and excessive collagen-I accumulation. The EMT and fibrotic effect of TGF-β1 dependent on acetylated C/EBPβ-mediated regulation of α-SMA gene activity. C/EBPβ acetylation may play a central role in pulmonary fibrosis.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6