TGF-β1 stimulates epithelial‑mesenchymal transition mediated by ADAM33

Fang,Liping; Wu,Jie; Huang,Tao; Zhang,Pengpeng; Xin,Xiaofeng; Shi,Yi

doi:10.3892/etm.2017.5478

TGF-β1 stimulates epithelial‑mesenchymal transition mediated by ADAM33

Authors:
- Liping Fang
- Jie Wu
- Tao Huang
- Pengpeng Zhang
- Xiaofeng Xin
- Yi Shi
View Affiliations

Affiliations: Department of Respiratory Medicine, Jinling Hospital, Nanjing, Jiangsu 210002, P.R. China, Cadre Department of Respiratory Medicine, Jinling Hospital, Nanjing, Jiangsu 210002, P.R. China, Department of Orthopedics, The First Affiliated Hospital of Xi'an Medical University, Xi'an, Shaanxi 710077, P.R. China
Published online on: November 10, 2017 https://doi.org/10.3892/etm.2017.5478
Pages: 985-992

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to determine the effects of transforming growth factor (TGF)‑β1 on disintegrin and metalloproteinase domain‑containing protein 33 (ADAM33) expression in airway epithelial cells in order to investigate the association between ADAM33 expression and TGF‑β1‑induced epithelial to mesenchymal transition (EMT), and to further explore the mechanisms underlying the role of ADAM33 in airway remodeling in asthma. The human bronchial epithelial cell line HBE was transfected with small interfering RNA targeting ADAM33 (siADAM33) and treated with different concentrations of TGF‑β1 (10, 20 or 30 ng/ml), while untransfected cells were used as controls. At 72 h after treatment, cellular morphology and immunohistochemical staining were observed under a microscope. The protein and mRNA expression levels of ADAM33 and the EMT markers E‑cadherin and vimentin were detected by western blot analysis and reverse‑transcription quantitative polymerase chain reaction, respectively. In addition, a correlation analysis of ADAM33 expression and E‑cadherin/vimentin expression was performed. A wound healing migration assay and a cell invasion assay were also performed. The results of the cellular morphology, migration and invasion studies suggested that TGF‑β1 treatment induced typical EMT changes in HBE cells. In addition, treatment with various concentrations of TGF‑β1 significantly increased the protein and mRNA expression levels of ADAM33 and vimentin compared with those in untreated cells. TGF‑β1 treatment also decreased the protein and mRNA expression levels of E‑cadherin in a dose‑dependent manner. By contrast, transfection with siADAM33 promoted the protein expression of E‑cadherin and decreased the protein expression of vimentin. Furthermore, ADAM33 and E‑cadherin expression levels exhibited a significant negative correlation, whereas ADAM33 and vimentin were positively correlated. In conclusion, the results suggested that TGF‑β1 enhances ADAM33 expression in airway epithelial cells, and that ADAM33 induces the EMT of airway epithelial cells, thus participating in airway remodeling in asthma.

View Figures

View References

1	Heijink IH, Postma DS, Noordhoek JA, Broekema M and Kapus A: House dust mite-promoted epithelial-to-mesenchymal transition in human bronchial epithelium. Am J Respir Cell Mol Biol. 42:69–79. 2010. View Article : Google Scholar : PubMed/NCBI
2	Hackett TL, Warner SM, Stefanowicz D, Shaheen F, Pechkovsky DV, Murray LA, Argentieri R, Kicic A, Stick SM, Bai TR and Knight DA: Induction of epithelial-mesenchymal transition in primary airway epithelial cells from patients with asthma by transforming growth factor-beta1. Am J Respir Crit Care Med. 180:122–133. 2009. View Article : Google Scholar : PubMed/NCBI
3	Hackett TL: Epithelial-mesenchymal transition in the pathophysiology of airway remodelling in asthma. Curr Opin Allergy Clin Immunol. 12:53–59. 2012. View Article : Google Scholar : PubMed/NCBI
4	Johnson JR, Roos A, Berg T, Nord M and Fuxe J: Chronic respiratory aeroallergen exposure in mice induces epithelial-mesenchymal transition in the large airway. PLoS One. 6:e161752011. View Article : Google Scholar : PubMed/NCBI
5	Wang JH, Sun LH, Huang SK and Chen AH: Effect of co-culturing human primary basic fibroblasts with respiratory syncytial virus-infected 16-HBE cells. Genet Mol Res. 15:150173392016.
6	Ko H, So Y, Jeon H, Jeong MH, Choi HK, Ryu SH, Lee SW, Yoon HG and Choi KC: TGF-β1-induced epithelial-mesenchymal transition and acetylation of Smad2 and Smad3 are negatively regulated by EGCG in human A549 lung cancer cells. Cancer Lett. 335:205–213. 2013. View Article : Google Scholar : PubMed/NCBI
7	Van Eerdewegh P, Little RD, Dupuis J, Del Mastro RG, Falls K, Simon J, Torrey D, Pandit S, McKenny J, Braunschweiger K, et al: Association of the ADAM33 gene with asthma and bronchial hyperresponsiveness. Nature. 418:426–430. 2002. View Article : Google Scholar : PubMed/NCBI
8	Lee JY, Park SW, Chang HK, Kim HY, Rhim T, Lee JH, Jang AS, Koh ES and Park CS: A disintegrin and metalloproteinase 33 protein in patients with asthma: Relevance to airflow limitation. Am J Respir Crit Care Med. 173:729–735. 2006. View Article : Google Scholar : PubMed/NCBI
9	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
10	Holgate ST: ADAM metallopeptidase domain 33 (ADAM33): Identification and role in airways disease. Drug News Perspect. 23:381–387. 2010.PubMed/NCBI
11	Xiao J, Han J, Wang X, Hua D, Su D, Bao Y and Lv F: Association of ADAM33 gene with susceptibility to COPD in Tibetan population of China. Mol Biol Rep. 38:4941–4945. 2011. View Article : Google Scholar : PubMed/NCBI
12	Sunadome H, Matsumoto H, Petrova G, Kanemitsu Y, Tohda Y, Horiguchi T, Kita H, Kuwabara K, Tomii K, Otsuka K, et al: IL4Rα and ADAM33 as genetic markers in asthma exacerbations and type-2 inflammatory endotype. Clin Exp Allergy. 47:998–1006. 2017. View Article : Google Scholar : PubMed/NCBI
13	Ito I, Laporte JD, Fiset PO, Asai K, Yamauchi Y, Martin JG and Hamid Q: Downregulation of a disintegrin and metalloproteinase 33 by IFN-gamma in human airway smooth muscle cells. J Allergy Clin Immunol. 119:89–97. 2007. View Article : Google Scholar : PubMed/NCBI
14	Jie Z, Jin M, Cai Y, Bai C, Shen Y, Yuan Z, Hu Y and Holgate S: The effects of Th2 cytokines on the expression of ADAM33 in allergen-induced chronic airway inflammation. Respir Physiol Neurobiol. 168:289–294. 2009. View Article : Google Scholar : PubMed/NCBI
15	Tripathi P, Awasthi S, Husain N, Prasad R and Mishra V: Increased expression of ADAM33 protein in asthmatic patients as compared to non-asthmatic controls. Indian J Med Res. 137:507–514. 2013.PubMed/NCBI
16	Foley SC, Mogas AK, Olivenstein R, Fiset PO, Chakir J, Bourbeau J, Ernst P, Lemière C, Martin JG and Hamid Q: Increased expression of ADAM33 and ADAM8 with disease progression in asthma. J Allergy Clin Immunol. 119:863–871. 2007. View Article : Google Scholar : PubMed/NCBI
17	Lin F, Song A, Wu J, Jiang X, Long J, Chen J, Duan Y, Shi Y and Deng L: ADAM33 protein expression and the mechanics of airway smooth muscle cells and highly correlated in ovalbumin-sensitized rats. Mol Med Rep. 8:1209–1215. 2013. View Article : Google Scholar : PubMed/NCBI
18	Haitchi HM, Powell RM, Shaw TJ, Howarth PH, Wilson SJ, Wilson DI, Holgate ST and Davies DE: ADAM33 expression in asthmatic airways and humanembryonic lungs. Am J Respir Crit Care Med. 171:958–965. 2005. View Article : Google Scholar : PubMed/NCBI
19	Yang Y, Wicks J, Haitchi HM, Powell RM, Manuyakorn W, Howarth PH, Holgate ST and Davies DE: Regulation of a disintegrin and metalloprotease-33 expression by transforming growth factor-β. Am J Respir Cell Mol Biol. 46:633–640. 2012. View Article : Google Scholar : PubMed/NCBI
20	Zhang M, Zhang Z, Pan HY, Wang DX, Deng ZT and Ye XL: TGF-beta1 induces human bronchial epithelial cell-to-mesenchymal transition in vitro. Lung. 187:187–194. 2009. View Article : Google Scholar : PubMed/NCBI
21	Kamitani S, Yamauchi Y, Kawasaki S, Takami K, Takizawa H, Nagase T and Kohyama T: Simultaneous stimulation with TGF-β1 and TNF-α induces epithelial mesenchymal transition in bronchial epithelial cells. Int Arch Allergy Immunol. 155:119–128. 2011. View Article : Google Scholar : PubMed/NCBI
22	Yang ZC, Yi MJ, Ran N, Wang C, Fu P, Feng XY, Xu L and Qu ZH: Transforming growth factor-β1 induces bronchial epithelial cells to mesenchymal transition by activating the Snail pathway and promotes airway remodeling in asthma. Mol Med Rep. 8:1663–1668. 2013. View Article : Google Scholar : PubMed/NCBI