Hypoxia-inducible factor-1α regulates epithelial-to-mesenchymal transition in paraquat-induced pulmonary fibrosis by activating lysyl oxidase

Lu,Jian; Qian,Yongbing; Jin,Wei; Tian,Rui; Zhu,Yong; Wang,Jinfeng; Meng,Xiaoxiao; Wang,Ruilan

doi:10.3892/etm.2017.5677

Hypoxia-inducible factor-1α regulates epithelial-to-mesenchymal transition in paraquat-induced pulmonary fibrosis by activating lysyl oxidase

Authors:
- Jian Lu
- Yongbing Qian
- Wei Jin
- Rui Tian
- Yong Zhu
- Jinfeng Wang
- Xiaoxiao Meng
- Ruilan Wang
View Affiliations

Affiliations: Department of Critical Care Medicine, Shanghai General Hospital of Nanjing Medical University, Shanghai 201620, P.R. China, Department of Critical Care Medicine, Shanghai General Hospital, Shanghai Jiaotong University, School of Medicine, Shanghai 201620, P.R. China
Published online on: December 22, 2017 https://doi.org/10.3892/etm.2017.5677
Pages: 2287-2294
Copyright: © Lu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Pulmonary fibrosis (PF) is one of the most prevalent causes of death following paraquat (PQ) poisoning. As demonstrated in previous studies by the present authors, epithelial-to-mesenchymal transition (EMT) is associated with PQ‑induced PF. In addition, hypoxia‑inducible factor‑1α (HIF‑1α) and lysyl oxidase (LOX) promote EMT following PQ poisoning. However, the association between HIF‑1α‑ and LOX‑mediated regulation of EMT remains unclear. The present study investigated the association between HIF‑1α and LOX with regard to PQ‑induced EMT. A549 and RLE‑6TN cells were treated with PQ, and HIF‑1α and LOX expression was silenced with short interfering RNAs. Changes in the expression of HIF‑1α, LOX, β‑catenin and EMT‑related makers were detected using real‑time quantitative polymerase chain reaction, immunofluorescence, and western blotting. HIF‑1α and LOX were associated with PQ‑induced EMT, and their expression levels were significantly increased (P<0.05). LOX expression was significantly decreased following PQ poisoning when HIF‑1α expression was inhibited (P<0.05). However, the level of HIF‑1α did not change significantly when LOX was silenced. The expression level of β‑catenin and the degree of EMT were significantly decreased following HIF‑1α and LOX silencing in both cell lines (P<0.05). The association between HIF‑1α and LOX in regulating EMT during PQ‑induced PF may be unidirectional. HIF‑1α may regulate PQ‑induced EMT through the LOX/β‑catenin pathway.

View Figures

View References

1	Gil HW, Hong JR, Jang SH and Hong SY: Diagnostic and therapeutic approach for acute paraquat intoxication. J Korean Med Sci. 29:1441–1449. 2014. View Article : Google Scholar : PubMed/NCBI
2	Xu L, Xu J and Wang Z: Molecular mechanisms of paraquat-induced acute lung injury: A current review. Drug Chem Toxicol. 37:130–134. 2014. View Article : Google Scholar : PubMed/NCBI
3	Dinis-Oliveira RJ, Duarte JA, Sanchez-Navarro A, Remiao F, Bastos ML and Carvalho F: Paraquat poisonings: Mechanisms of lung toxicity, clinical features, and treatment. Crit Rev Toxicol. 38:13–71. 2008. View Article : Google Scholar : PubMed/NCBI
4	Nieto MA, Huang RY, Jackson RA and Thiery JP: EMT: 2016. Cell. 166:21–45. 2016. View Article : Google Scholar : PubMed/NCBI
5	Stone RC, Pastar I, Ojeh N, Chen V, Liu S, Garzon KI and Tomic-Canic M: Epithelial-mesenchymal transition in tissue repair and fibrosis. Cell Tissue Res. 365:495–506. 2016. View Article : Google Scholar : PubMed/NCBI
6	Bartis D, Mise N, Mahida RY, Eickelberg O and Thickett DR: Epithelial-mesenchymal transition in lung development and disease: Does it exist and is it important? Thorax. 69:760–765. 2014. View Article : Google Scholar : PubMed/NCBI
7	Kage H and Borok Z: EMT and interstitial lung disease: A mysterious relationship. Curr Opin Pulm Med. 18:517–523. 2012.PubMed/NCBI
8	Wang J, Zhu Y, Tan J, Meng X, Xie H and Wang R: Lysyl oxidase promotes epithelial-to-mesenchymal transition during paraquat-induced pulmonary fibrosis. Mol Biosyst. 12:499–507. 2016. View Article : Google Scholar : PubMed/NCBI
9	Zhu Y, Tan J, Xie H, Wang J, Meng X and Wang R: HIF-1alpha regulates EMT via the Snail and beta-catenin pathways in paraquat poisoning-induced early pulmonary fibrosis. J Cell Mol Med. 20:688–697. 2016. View Article : Google Scholar : PubMed/NCBI
10	Masoud GN and Li W: HIF-1alpha pathway: Role, regulation and intervention for cancer therapy. Acta Pharm Sin B. 5:378–389. 2015. View Article : Google Scholar : PubMed/NCBI
11	Balamurugan K: HIF-1 at the crossroads of hypoxia, inflammation, and cancer. Int J Cancer. 138:1058–1066. 2016. View Article : Google Scholar : PubMed/NCBI
12	Zhou G, Dada LA, Wu M, Kelly A, Trejo H, Zhou Q, Varga J and Sznajder JI: Hypoxia-induced alveolar epithelial-mesenchymal transition requires mitochondrial ROS and hypoxia-inducible factor 1. Am J Physiol Lung Cell Mol Physiol. 297:L1120–1130. 2009. View Article : Google Scholar : PubMed/NCBI
13	Darby IA and Hewitson TD: Hypoxia in tissue repair and fibrosis. Cell Tissue Res. 365:553–562. 2016. View Article : Google Scholar : PubMed/NCBI
14	Wang RL, Tang X, Wu X, Xu R, Yu KL and Xu K: The relationship between HIF-1α expression and the early lung fibrosis in rats with acute paraquat poisoning. Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 30:273–277. 2012.(In Chinese). PubMed/NCBI
15	Cox TR, Bird D, Baker AM, Barker HE, Ho MW, Lang G and Erler JT: LOX-mediated collagen crosslinking is responsible for fibrosis-enhanced metastasis. Cancer Res. 73:1721–1732. 2013. View Article : Google Scholar : PubMed/NCBI
16	Lopez B, Gonzalez A, Hermida N, Valencia F, de Teresa E and Diez J: Role of lysyl oxidase in myocardial fibrosis: From basic science to clinical aspects. Am J Physiol Heart Circ Physiol. 299:H1–9. 2010. View Article : Google Scholar : PubMed/NCBI
17	Deng S, Jin T, Zhang L, Bu H and Zhang P: Mechanism of tacrolimus-induced chronic renal fibrosis following transplantation is regulated by ox-LDL and its receptor, LOX-1. Mol Med Rep. 14:4124–4134. 2016. View Article : Google Scholar : PubMed/NCBI
18	Cheng T, Liu Q, Zhang R, Zhang Y, Chen J, Yu R and Ge G: Lysyl oxidase promotes bleomycin-induced lung fibrosis through modulating inflammation. J Mol Cell Biol. 6:506–515. 2014. View Article : Google Scholar : PubMed/NCBI
19	Erler JT, Bennewith KL, Nicolau M, Dornhöfer N, Kong C, Le QT, Chi JT, Jeffrey SS and Giaccia AJ: Lysyl oxidase is essential for hypoxia-induced metastasis. Nature. 440:1222–1226. 2006. View Article : Google Scholar : PubMed/NCBI
20	Ji F, Wang Y, Qiu L, Li S, Zhu J, Liang Z, Wan Y and Di W: Hypoxia inducible factor 1α-mediated LOX expression correlates with migration and invasion in epithelial ovarian cancer. Int J Oncol. 42:1578–1588. 2013. View Article : Google Scholar : PubMed/NCBI
21	Pez F, Dayan F, Durivault J, Kaniewski B, Aimond G, Le Provost GS, Deux B, Clézardin P, Sommer P, Pouysségur J and Reynaud C: The HIF-1-inducible lysyl oxidase activates HIF-1 via the Akt pathway in a positive regulation loop and synergizes with HIF-1 in promoting tumor cell growth. Cancer Res. 71:1647–1657. 2011. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-tie quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
23	Lamouille S, Xu J and Derynck R: Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 15:178–196. 2014. View Article : Google Scholar : PubMed/NCBI
24	Elvidge GP, Glenny L, Appelhoff RJ, Ratcliffe PJ, Ragoussis J and Gleadle JM: Concordant regulation of gene expression by hypoxia and 2-oxoglutarate-dependent dioxygenase inhibition: The role of HIF-1alpha, HIF-2alpha, and other pathways. J Biol Chem. 281:15215–15226. 2006. View Article : Google Scholar : PubMed/NCBI
25	Goto TM, Arima Y, Nagano O and Saya H: Lysyl oxidase is induced by cell density-mediated cell cycle suppression via RB-E2F1-HIF-1α axis. Cell Struct Funct. 38:9–14. 2013. View Article : Google Scholar : PubMed/NCBI
26	Yang X, Li S, Li W, Chen J, Xiao X, Wang Y, Yan G and Chen L: Inactivation of lysyl oxidase by β-aminopropionitrile inhibits hypoxia-induced invasion and migration of cervical cancer cells. Oncol Rep. 29:541–548. 2013. View Article : Google Scholar : PubMed/NCBI
27	Wang Y, Ma J, Shen H, Wang C, Sun Y, Howell SB and Lin X: Reactive oxygen species promote ovarian cancer progression via the HIF-1α/LOX/E-cadherin pathway. Oncol Rep. 32:2150–2158. 2014. View Article : Google Scholar : PubMed/NCBI
28	Reynaud C, Ferreras L, Di Mauro P, Kan C, Croset M, Bonnelye E, Pez F, Thomas C, Aimond G, Karnoub AE, et al: Lysyl oxidase is a strong determinant of tumor cell colonization in bone. Cancer Res. 77:268–278. 2017. View Article : Google Scholar : PubMed/NCBI
29	Schietke R, Warnecke C, Wacker I, Schödel J, Mole DR, Campean V, Amann K, Goppelt-Struebe M, Behrens J, Eckardt KU and Wiesener MS: The lysyl oxidases LOX and LOXL2 are necessary and sufficient to repress E-cadherin in hypoxia: Insights into cellular transformation processes mediated by HIF-1. J Biol Chem. 285:6658–6669. 2010. View Article : Google Scholar : PubMed/NCBI
30	Zeisberg M and Neilson EG: Biomarkers for epithelial-mesenchymal transitions. J Clin Invest. 119:1429–1437. 2009. View Article : Google Scholar : PubMed/NCBI
31	Xu L, Cui WH, Zhou WC, Li DL, Li LC, Zhao P, Mo XT, Zhang Z and Gao J: Activation of Wnt/β-catenin signalling is required for TGF-β/Smad2/3 signalling during myofibroblast proliferation. J Cell Mol Med. 21:1545–1554. 2017. View Article : Google Scholar : PubMed/NCBI
32	Ji S, Deng H, Jin W, Yan P, Wang R, Pang L, Zhou J, Zhang J, Chen X, Zhao X and Shen J: Beta-catenin participates in dialysate-induced peritoneal fibrosis via enhanced peritoneal cell epithelial-to-mesenchymal transition. FEBS Open Bio. 7:265–273. 2017. View Article : Google Scholar : PubMed/NCBI
33	Wang X, Dai W, Wang Y, Gu Q, Yang D and Zhang M: Blocking the Wnt/β-catenin pathway by lentivirus-mediated short hairpin RNA targeting β-catenin gene suppresses silica-induced lung fibrosis in mice. Int J Environ Res Public Health. 12:10739–10754. 2015. View Article : Google Scholar : PubMed/NCBI
34	Martinez-Martinez E, Ibarrola J, Calvier L, Fernandez-Celis A, Leroy C, Cachofeiro V, Rossignol P and Lopez-Andres N: Galectin-3 blockade reduces renal fibrosis in two normotensive experimental models of renal damage. PLoS One. 11:e01662722016. View Article : Google Scholar : PubMed/NCBI
35	Hu BL, Shi C, Lei RE, Lu DH, Luo W, Qin SY, Zhou Y and Jianga HX: Interleukin-22 ameliorates liver fibrosis through miR-200a/beta-catenin. Sci Rep. 6:364362016. View Article : Google Scholar : PubMed/NCBI
36	Lin JC, Kuo WW, Baskaran R, Chen MC, Ho TJ, Chen RJ, Chen YF, Vijaya Padma V, Lay IS and Huang CY: Enhancement of beta-catenin in cardiomyocytes suppresses survival protein expression but promotes apoptosis and fibrosis. Cardiol J. 24:195–205. 2017. View Article : Google Scholar : PubMed/NCBI
37	Anastas JN and Moon RT: WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer. 13:11–26. 2013. View Article : Google Scholar : PubMed/NCBI
38	Xie H, Tan JT, Wang RL, Meng XX, Tang X and Gao S: Expression and significance of HIF-1alpha in pulmonary fibrosis induced by paraquat. Exp Biol Med (Maywood). 238:1062–1068. 2013. View Article : Google Scholar : PubMed/NCBI