Kinase inhibitors fail to induce mesenchymal-epithelial transition in fibroblasts from fibrotic lung tissue

Xu,Xuefeng; Wan,Xuan; Geng,Jing; Li,Fei; Wang,Chen; Dai,Huaping

doi:10.3892/ijmm.2013.1415

Kinase inhibitors fail to induce mesenchymal-epithelial transition in fibroblasts from fibrotic lung tissue

Authors:
- Xuefeng Xu
- Xuan Wan
- Jing Geng
- Fei Li
- Chen Wang
- Huaping Dai
View Affiliations

Affiliations: Department of Respiratory and Critical Care Medicine, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, P.R. China, Beijing Key Laboratory of Respiratory and Pulmonary Circulation Disorders, Beijing Institute of Respiratory Medicine, Beijing 100020, P.R. China
Published online on: June 11, 2013 https://doi.org/10.3892/ijmm.2013.1415
Pages: 430-438

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

Epithelial-mesenchymal transition (EMT) has been considered to be involved in idiopathic pulmonary fibrosis (IPF). However, the EMT process in vivo is much more complex and controversial. Studies regarding the opposite process, mesenchymal-epithelial transition (MET) in fibroblasts, are limited. Therefore, the aim of this study was to verify the involvement of the transforming growth factor (TGF)-β1-dependent EMT network in the process of pulmonary fibrosis and to explore the possibility of MET. Fibrotic lung tissues were obtained from patients with IPF with histological evidence of usual interstitial pneumonia at the time of surgical lung biopsy. For the controls, histologically normal lung tissues were obtained from patients with primary spontaneous pneumothorax at the time of thoracoscopy with stapling of any air leaks. Real-time RT-PCR and western blot analysis revealed that the mRNA and protein levels of TGF-β1, TGF-β1 receptor type I/II/III (TβRI/II/III), Smad2/3/4 and Snail1/2 were significantly upregulated in the fibrotic lung tissue. Inhibitors of various kinases implicated in EMT, including TGF-β1/Smad, Rho kinase (ROCK), p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun NH-terminal kinase (JNK) were used to determine the MET potential in fibroblasts from fibrotic lung tissue. Western blot analysis or indirect immunofluorescence staining revealed that Smad inhibitor, as well as other kinase inhibitors failed to induce the MET process, determined by cellular morphology and protein markers. Our data suggest that the MET process may not be the exact reversal of EMT. In addition to using kinase inhibitors, other intervention measures should be used to explore the possibility of the MET process in fibroblasts from fibrotic lung tissue.

View Figures

View References

1	King TE Jr, Pardo A and Selman M: Idiopathic pulmonary fibrosis. Lancet. 378:1949–1961. 2011. View Article : Google Scholar
2	Selman M, Thannickal VJ, Pardo A, Zisman DA, Martinez FJ and Lynch JP III: Idiopathic pulmonary fibrosis: pathogenesis and therapeutic approaches. Drugs. 64:405–430. 2004. View Article : Google Scholar : PubMed/NCBI
3	Phan SH: Biology of fibroblasts and myofibroblasts. Proc Am Thorac Soc. 5:334–337. 2008. View Article : Google Scholar : PubMed/NCBI
4	Lee K and Nelson CM: New insights into the regulation of epithelial-mesenchymal transition and tissue fibrosis. Int Rev Cell Mol Biol. 294:171–221. 2012. View Article : Google Scholar : PubMed/NCBI
5	Coward WR, Saini G and Jenkins G: The pathogenesis of idiopathic pulmonary fibrosis. Ther Adv Respir Dis. 4:367–388. 2010. View Article : Google Scholar : PubMed/NCBI
6	Gharaee-Kermani M, Hu B, Phan SH and Gyetko MR: Recent advances in molecular targets and treatment of idiopathic pulmonary fibrosis: focus on TGFbeta signaling and the myofibroblast. Curr Med Chem. 16:1400–1417. 2009. View Article : Google Scholar : PubMed/NCBI
7	Chapman HA: Epithelial-mesenchymal interactions in pulmonary fibrosis. Annu Rev Physiol. 73:413–435. 2011. View Article : Google Scholar : PubMed/NCBI
8	Kalluri R and Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 119:1420–1428. 2009. View Article : Google Scholar : PubMed/NCBI
9	Acloque H, Adams MS, Fishwick K, Bronner-Fraser M and Nieto MA: Epithelial-mesenchymal transitions: the importance of changing cell state in development and disease. J Clin Invest. 119:1438–1449. 2009. View Article : Google Scholar : PubMed/NCBI
10	Thiery JP, Acloque H, Huang RY and Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell. 139:871–890. 2009. View Article : Google Scholar : PubMed/NCBI
11	Kalluri R and Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest. 112:1776–1784. 2003. View Article : Google Scholar : PubMed/NCBI
12	Lan HY and Chung AC: Transforming growth factor-beta and Smads. Contrib Nephrol. 170:75–82. 2011. View Article : Google Scholar : PubMed/NCBI
13	Pohlers D, Brenmoehl J, Loffler I, Müller CK, Leipner C, Schultze-Mosgau S, Stallmach A, Kinne RW and Wolf G: TGF-beta and fibrosis in different organs - molecular pathway imprints. Biochim Biophys Acta. 1792:746–756. 2009. View Article : Google Scholar : PubMed/NCBI
14	Willis BC, Liebler JM, Luby-Phelps K, Nicholson AG, Crandall ED, du Bois RM and Borok Z: Induction of epithelial-mesenchymal transition in alveolar epithelial cells by transforming growth factor-beta1: potential role in idiopathic pulmonary fibrosis. Am J Pathol. 166:1321–1332. 2005. View Article : Google Scholar
15	Kasai H, Allen JT, Mason RM, Kamimura T and Zhang Z: TGF-beta1 induces human alveolar epithelial to mesenchymal cell transition (EMT). Respir Res. 6:562005. View Article : Google Scholar : PubMed/NCBI
16	Fragiadaki M and Mason RM: Epithelial-mesenchymal transition in renal fibrosis - evidence for and against. Int J Exp Pathol. 92:143–150. 2011. View Article : Google Scholar : PubMed/NCBI
17	American Thoracic Society; European Respiratory Society. American Thoracic Society/European Respiratory Society International Multidisciplinary Consensus Classification of the Idiopathic Interstitial Pneumonias. This joint statement of the American Thoracic Society (ATS), and the European Respiratory Society (ERS) was adopted by the ATS board of directors, June 2001 and by the ERS Executive Committee, June 2001. Am J Respir Crit Care Med. 165:277–304. 2002.
18	Hutchison N, Hendry BM and Sharpe CC: Rho isoforms have distinct and specific functions in the process of epithelial to mesenchymal transition in renal proximal tubular cells. Cell Signal. 21:1522–1531. 2009. View Article : Google Scholar : PubMed/NCBI
19	Lv ZM, Wang Q, Wan Q, Lin JG, Hu MS, Liu YX and Wang R: The role of the p38 MAPK signaling pathway in high glucose-induced epithelial-mesenchymal transition of cultured human renal tubular epithelial cells. PLoS One. 6:e228062011. View Article : Google Scholar : PubMed/NCBI
20	Kolosova I, Nethery D and Kern JA: Role of Smad2/3 and p38 MAP kinase in TGF-β1-induced epithelial-mesenchymal transition of pulmonary epithelial cells. J Cell Physiol. 226:1248–1254. 2011.
21	van der Velden JL, Guala AS, Leggett SE, Sluimer J, Badura EC and Janssen-Heininger YM: Induction of a mesenchymal expression program in lung epithelial cells by wingless protein (Wnt)/β-catenin requires the presence of c-Jun N-terminal kinase-1 (JNK1). Am J Respir Cell Mol Biol. 47:306–314. 2012.PubMed/NCBI
22	Selman M, Pardo A and Kaminski N: Idiopathic pulmonary fibrosis: aberrant recapitulation of developmental programs? PLoS Med. 5:e622008. View Article : Google Scholar : PubMed/NCBI
23	Lomas NJ, Watts KL, Akram KM, Forsyth NR and Spiteri MA: Idiopathic pulmonary fibrosis: immunohistochemical analysis provides fresh insights into lung tissue remodelling with implications for novel prognostic markers. Int J Clin Exp Pathol. 5:58–71. 2012.
24	Yamada M, Kuwano K, Maeyama T, Hamada N, Yoshimi M, Nakanishi Y and Kasper M: Dual-immunohistochemistry provides little evidence for epithelial-mesenchymal transition in pulmonary fibrosis. Histochem Cell Biol. 129:453–462. 2008. View Article : Google Scholar : PubMed/NCBI
25	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
26	Kim JH, Jang YS, Eom KS, Hwang YI, Kang HR, Jang SH, Kim CH, Park YB, Lee MG, Hyun IG, Jung KS and Kim DG: Transforming growth factor beta1 induces epithelial-to-mesenchymal transition of A549 cells. J Korean Med Sci. 22:898–904. 2007. View Article : Google Scholar : PubMed/NCBI
27	Fernando RI, Castillo MD, Litzinger M, Hamilton DH and Palena C: IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells. Cancer Res. 71:5296–5306. 2011. View Article : Google Scholar : PubMed/NCBI
28	Das S, Becker BN, Hoffmann FM and Mertz JE: Complete reversal of epithelial to mesenchymal transition requires inhibition of both ZEB expression and the Rho pathway. BMC Cell Biol. 10:942009. View Article : Google Scholar : PubMed/NCBI
29	Ramos C, Becerril C, Montano M, Garcia-De-Alba C, Ramirez R, Checa M, Pardo A and Selman M: FGF-1 reverts epithelial-mesenchymal transition induced by TGF-{beta}1 through MAPK/ERK kinase pathway. Am J Physiol Lung Cell Mol Physiol. 299:L222–L231. 2010.PubMed/NCBI
30	Dudas PL, Argentieri RL and Farrell FX: BMP-7 fails to attenuate TGF-beta1-induced epithelial-to-mesenchymal transition in human proximal tubule epithelial cells. Nephrol Dial Transplant. 24:1406–1416. 2009. View Article : Google Scholar : PubMed/NCBI