Overexpression of the hyperplasia suppressor gene inactivates airway fibroblasts obtained from a rat model of chronic
obstructive pulmonary disease by inhibiting the Wnt signaling pathway

Ge,Zhenghang; Yang,Yi; Zhou,Xun; Zhang,Jun; Li,Bo; Wang,Xinxing; Luo,Xian

doi:10.3892/mmr.2019.10504

Overexpression of the hyperplasia suppressor gene inactivates airway fibroblasts obtained from a rat model of chronic obstructive pulmonary disease by inhibiting the Wnt signaling pathway

Authors:
- Zhenghang Ge
- Yi Yang
- Xun Zhou
- Jun Zhang
- Bo Li
- Xinxing Wang
- Xian Luo
View Affiliations

Affiliations: Department of Respiratory Medicine, The Second Affiliated Hospital of Guizhou College of Traditional Chinese Medicine, Guiyang, Guizhou 550003, P.R. China, Department of Research and Teaching, The Second Affiliated Hospital of Guizhou College of Traditional Chinese Medicine, Guiyang, Guizhou 550003, P.R. China
Published online on: July 15, 2019 https://doi.org/10.3892/mmr.2019.10504
Pages: 2754-2762
Copyright: © Ge 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

The present study aimed to investigate the effects of hyperplasia suppressor gene (HSG) overexpression on the activation of airway fibroblasts in a rat model of chronic obstructive pulmonary disease (COPD) and assess the underlying molecular mechanisms. The rat model of COPD was established by injection of papain and confirmed by hematoxylin and eosin staining. Airway fibroblasts were identified using immunofluorescence, and HSG expression was facilitated by an HSG vector. Cell viability, apoptosis and the levels of matrix metallopeptidase‑9 (MMP‑9), platelet‑derived growth factor (PDGF), and transforming growth factor‑β1 (TGF‑β1) were measured via Cell Counting Kit‑8, flow cytometry and ELISA analyses, respectively, and potential mechanisms were detected by reverse transcription‑quantitative polymerase chain reaction and western blotting. Airway fibroblasts from COPD rats were isolated and identified based on vimentin expression. Compared with the control group, HSG overexpression reduced cell viability, promoted apoptosis, and reduced the protein levels of TGF‑β1, MMP‑9 and PDGF. Additionally, HSG overexpression reduced β‑catenin and Ras homology family member A (RhoA) expression at both the mRNA and protein levels. Conversely, Wnt signaling pathway agonists lithium chloride (LiCl) and 4‑ethyl‑5,6‑dihydro‑5‑methyl‑(1,3)dioxolo(4,5‑j)phenanthridine (HLY78), significantly reduced the effects of HSG overexpression (P<0.05 vs. HSG). Cell viability in the HSG + LiCl and HSG + HLY78 groups was increased, whereas apoptosis was reduced compared with HSG treatment alone. The protein levels of TGF‑β1, MMP‑9 and PDGF were also decreased in the HSG + LiCl and HSG + HLY78 groups compared with the HSG group (P<0.05). Furthermore, the expression of β‑catenin and RhoA was higher in the HSG + LiCl and HSG + HLY78 groups compared with the HSG group (P<0.05). Collectively, the results indicated that HSG overexpression inactivated airway fibroblasts from COPD by inhibiting the Wnt signaling pathway.

View Figures

View References

1	Yu ZW, Xu YQ, Zhang XJ, Pan JR, Xiang HX, Gu XH, Ji SB and Qian J: Mutual regulation between miR-21 and the TGFβ/Smad signaling pathway in human bronchial fibroblasts promotes airway remodeling. J Asthma. 56:341–349. 2019. View Article : Google Scholar : PubMed/NCBI
2	Lai T, Tian B, Cao C, Hu Y, Zhou J, Wang Y, Wu Y, Li Z, Xu X, Zhang M, et al: HDAC2 suppresses IL17A-mediated airway remodeling in human and experimental modeling of COPD. Chest. 153:863–875. 2018. View Article : Google Scholar : PubMed/NCBI
3	Ricciardolo FLM, Folkerts G, Folino A and Mognetti B: Bradykinin in asthma: Modulation of airway inflammation and remodelling. Eur J Pharmacol. 827:181–188. 2018. View Article : Google Scholar : PubMed/NCBI
4	Eapen MS, Myers S, Lu W, Tanghe C, Sharma P and Sohal SS: sE-cadherin and sVE-cadherin indicate active epithelial/endothelial to mesenchymal transition (EMT and EndoMT) in smokers and COPD: Implications for new biomarkers and therapeutics. Biomarkers. 23:709–711. 2018. View Article : Google Scholar : PubMed/NCBI
5	Mahmood MQ, Reid D, Ward C, Muller HK, Knight DA, Sohal SS and Walters EH: Transforming growth factor (TGF) β₁ and Smad signalling pathways: A likely key to EMT-associated COPD pathogenesis. Respirology. 22:133–140. 2017. View Article : Google Scholar : PubMed/NCBI
6	Sun ZQ, Chen G, Guo Q, Li HF and Wang Z: In vivo and in vitro effects of hyperplasia suppressor gene on the proliferation and apoptosis of lung adenocarcinoma A549 cells. Biosci Reports. 38:BSR201803912018. View Article : Google Scholar
7	Luo L, Gong YQ, Qi X, Lai W, Lan H and Luo Y: Effect of tumor suppressor PTEN gene on apoptosis and cell cycle of human airway smooth muscle cells. Mol Cell Biochem. 375:1–9. 2013.PubMed/NCBI
8	Jiang GJ, Han M, Zheng B and Wen JK: Hyperplasia suppressor gene associates with smooth muscle alpha-actin and is involved in the redifferentiation of vascular smooth muscle cells. Heart Vessels. 21:315–320. 2006. View Article : Google Scholar : PubMed/NCBI
9	Guo YH, Li Q, Yu HY and Gao W: Hyperplasia suppressor gene induces vascular smooth muscle cell apoptosis. Beijing Da Xue Xue Bao Yi Xue Ban. 39:394–398. 2007.(In Chinese). PubMed/NCBI
10	Wu L, Li Z, Zhang Y, Zhang P, Zhu X, Huang J, Ma T, Lu T, Song Q, Li Q, et al: Adenovirus-expressed human hyperplasia suppressor gene induces apoptosis in cancer cells. Mol Cancer Ther. 7:222–232. 2008. View Article : Google Scholar : PubMed/NCBI
11	Zhang Y, Du Q, Qiu XY, Tian XX and Fang WG: Over expression of hyperplasia suppressor gene inhibits the malignant phenotype of breast cancer cell. Zhonghua Bing Li Xue Za Zhi. 39:259–263. 2010.(In Chinese). PubMed/NCBI
12	Zheng-Xing GE, Bo LI, Zhou X and Chang LI: rHSG gene regulates airway fibroblast proliferation and apoptosis of COPD rats. Basic Clin Med. 33:1235–1241. 2013.
13	Villar J, Cabrera NE, Valladares F, Casula M, Flores C, Blanch L, Quilez ME, Santana-Rodríguez N, Kacmarek RM and Slutsky AS: Activation of the Wnt/β-catenin signaling pathway by mechanical ventilation is associated with ventilator-induced pulmonary fibrosis in healthy lungs. PLoS One. 6:e239142011. View Article : Google Scholar : PubMed/NCBI
14	Chilosi M, Poletti V, Zamò A, Lestani M, Montagna L, Piccoli P, Pedron S, Bertaso M, Scarpa A, Murer B, et al: Aberrant Wnt/beta-catenin pathway activation in idiopathic pulmonary fibrosis. Am J Pathol. 162:1495–1502. 2003. View Article : Google Scholar : PubMed/NCBI
15	Königshoff M, Balsara N, Pfaff EM, Kramer M, Chrobak I, Seeger W and Eickelberg O: Functional Wnt signaling is increased in idiopathic pulmonary fibrosis. PLoS One. 3:e21422008. View Article : Google Scholar : PubMed/NCBI
16	Peng Y, Zhang X, Ma Q, Yan R, Qin Y, Zhao Y, Cheng Y, Yang M, Wang Q, Feng X, et al: MiRNA-194 activates the Wnt/β-catenin signaling pathway in gastric cancer by targeting the negative Wnt regulator, SUFU. Cancer Lett. 385:117–127. 2017. View Article : Google Scholar : PubMed/NCBI
17	Clevers H and Nusse R: Wnt/β-catenin signaling and disease. Cell. 149:1192–1205. 2012. View Article : Google Scholar : PubMed/NCBI
18	Qu J, Yue L, Gao J and Yao H: Perspectives on Wnt signal pathway in the pathogenesis and therapeutics in chronic obstructive pulmonary disease. J Pharmacol Exp Ther. 369:473–480. 2019. View Article : Google Scholar : PubMed/NCBI
19	Tao H, Yang JJ, Shi KH and Li J: Wnt signaling pathway in cardiac fibrosis: New insights and directions. Metabolism. 65:30–40. 2016. View Article : Google Scholar : PubMed/NCBI
20	Nishikawa K, Osawa Y and Kimura K: Wnt/β-catenin signaling as a potential target for the treatment of liver cirrhosis using antifibrotic drugs. Int J Mol Sci. 19:E31032018. View Article : Google Scholar : PubMed/NCBI
21	Lewis CC, Chu HW, Westcott JY, Tucker A, Langmack EL, Sutherland ER and Kraft M: Airway fibroblasts exhibit a synthetic phenotype in severe asthma. J Allergy Clin Immunol. 115:534–540. 2005. View Article : Google Scholar : PubMed/NCBI
22	Song Z, Chen H, Xu W, Wu S and Zhu G: Basolateral amygdala calpain is required for extinction of contextual fear-memory. Neurobiol Learn Mem. 155:180–188. 2018. View Article : Google Scholar : PubMed/NCBI
23	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
24	Zhu G, Wang X, Wu S and Li Q: Involvement of activation of PI3K/Akt pathway in the protective effects of puerarin against MPP+-induced human neuroblastoma SH-SY5Y cell death. Neurochem Int. 60:400–408. 2012. View Article : Google Scholar : PubMed/NCBI
25	Aigon A and Billecocq S: Prevalence and impact on quality of life of urinary incontinence in an adult population with chronic obstructive pulmonary diseases, literature review. Prog Urol. 28:962–972. 2018.(In French). View Article : Google Scholar : PubMed/NCBI
26	Oshagbemi OA, Keene SJ, Driessen JHM, Jordan R, Wouters EFM, de Boer A, de Vries F and Franssen FME: Trends in moderate and severe exacerbations among COPD patients in the UK from 2005 to 2013. Respir Med. 144:1–6. 2018. View Article : Google Scholar : PubMed/NCBI
27	Moon JY, Leitao Filho FS, Shahangian K, Takiguchi H and Sin DD: Blood and sputum protein biomarkers for chronic obstructive pulmonary disease (COPD). Expert Rev Proteomics. 15:923–935. 2018. View Article : Google Scholar : PubMed/NCBI
28	Reuter S, Beckert H and Taube C: Take the Wnt out of the inflammatory sails: Modulatory effects of Wnt in airway diseases. Lab Invest. 96:177–185. 2016. View Article : Google Scholar : PubMed/NCBI
29	Bartel S, Carraro G, Alessandrini F, Krauss-Etschmann S, Ricciardolo FLM and Bellusci S: miR-142-3p is associated with aberrant WNT signaling during airway remodeling in asthma. Am J Physiol Lung Cell Mol Physiol. 315:L328–L333. 2018. View Article : Google Scholar : PubMed/NCBI
30	Royer PJ, Henrio K, Pain M, Loy J, Roux A, Tissot A, Lacoste P, Pison C, Brouard S and Magnan A; COLT consortium, : TLR3 promotes MMP-9 production in primary human airway epithelial cells through Wnt/β-catenin signaling. Respir Res. 18:2082017. View Article : Google Scholar : PubMed/NCBI
31	Koopmans T and Gosens R: Revisiting asthma therapeutics: Focus on WNT signal transduction. Drug Discov Today. 23:49–62. 2018. View Article : Google Scholar : PubMed/NCBI
32	Hussain M, Xu C, Lu M and Wu X, Tang L and Wu X: Wnt/β-catenin signaling links embryonic lung development and asthmatic airway remodeling. Biochim Biophys Acta Mol Basis Dis. 1863:3226–3242. 2017. View Article : Google Scholar : PubMed/NCBI
33	Vallée A, Lecarpentier Y, Guillevin R and Vallée JN: Interactions between TGF-β1, canonical WNT/β-catenin pathway and PPAR γ in radiation-induced fibrosis. Oncotarget. 8:90579–90604. 2017. View Article : Google Scholar : PubMed/NCBI
34	Si Y, Bai J, Wu J, Li Q, Mo Y, Fang R and Lai W: LncRNA PlncRNA1 regulates proliferation and differentiation of hair follicle stem cells through TGFbeta1mediated Wnt/betacatenin signal pathway. Mol Med Rep. 17:1191–1197. 2018.PubMed/NCBI
35	Ma F, Li W, Liu C, Li W, Yu H, Lei B, Ren Y, Li Z, Pang D and Qian C: MiR-23a promotes TGF-β1-induced EMT and tumor metastasis in breast cancer cells by directly targeting CDH1 and activating Wnt/β-catenin signaling. Oncotarget. 8:69538–69550. 2017.PubMed/NCBI
36	Li M, Yuan Y, Chen Q, Me R, Gu Q, Yu Y, Sheng M and Ke B: Expression of Wnt/β-catenin signaling pathway and its regulatory role in type I collagen with TGF-β1 in scleral fibroblasts from an experimentally induced myopia guinea pig model. J Ophthalmol. 2016:51265602016.PubMed/NCBI
37	Godinas L, Corhay JL, Henket M, Guiot J, Louis R and Moermans C: Increased production of TGF-β1 from sputum cells of COPD: Relationship with airway obstruction. Cytokine. 99:1–8. 2017. View Article : Google Scholar : PubMed/NCBI
38	Westergren-Thorsson G, Bagher M, Andersson-Sjoland A, Andersson-Sjöland A, Thiman L, Löfdahl CG, Hallgren O, Bjermer L and Larsson-Callerfelt AK: VEGF synthesis is induced by prostacyclin and TGF-beta in distal lung fibroblasts from COPD patients and control subjects: Implications for pulmonary vascular remodelling. Respirology. 23:68–75. 2018. View Article : Google Scholar : PubMed/NCBI
39	Chen H, Zhang R, Zheng Q, Yan X, Wu S and Chen Y: Impact of body mass index on long-term blood pressure variability: A cross-sectional study in a cohort of Chinese adults. BMC Public Health. 18:11932018. View Article : Google Scholar : PubMed/NCBI
40	Di Stefano A, Sangiorgi C, Gnemmi I, Casolari P, Brun P, Ricciardolo FLM, Contoli M, Papi A, Maniscalco P, Ruggeri P, et al: TGF-β signaling pathways in different compartments of the lower airways of patients with stable COPD. Chest. 153:851–862. 2018. View Article : Google Scholar : PubMed/NCBI
41	Zhdan VN, Potyazhenko MМ, Khaymenova GS, Lyulka NN, Dubrovinskaya TV and Ivanitsky IV: Intensifying approach to the therapy of patients with constellation of the diseases: Chronic obstructive pulmonary disease and osteoarthritis. Wiad Lek. 70:578–580. 2017.PubMed/NCBI
42	Zhao Y, Qiao X, Wang L, Tan TK, Zhao H, Zhang Y, Zhang J, Rao P, Cao Q, Wang Y, et al: Matrix metalloproteinase 9 induces endothelial-mesenchymal transition via Notch activation in human kidney glomerular endothelial cells. BMC Cell Biol. 17:212016. View Article : Google Scholar : PubMed/NCBI
43	Yang Z, Li K, Liang Q, Zheng G, Zhang S, Lao X, Liang Y and Liao G: Elevated hydrostatic pressure promotes ameloblastoma cell invasion through upregulation of MMP-2 and MMP-9 expression via Wnt/β-catenin signalling. J Oral Pathol Med. 47:836–846. 2018. View Article : Google Scholar : PubMed/NCBI
44	Lu J, Zhu Y, Feng W, Pan Y, Li S, Han D, Liu L, Xie X, Wang G and Li M: Platelet-derived growth factor mediates interleukin-13-induced collagen I production in mouse airway fibroblasts. J Biosci. 39:693–700. 2014. View Article : Google Scholar : PubMed/NCBI
45	Song S, Zhang M, Yi Z, Zhang H, Shen T, Yu X, Zhang C, Zheng X, Yu L, Ma C, et al: The role of PDGF-B/TGF-β1/neprilysin network in regulating endothelial-to-mesenchymal transition in pulmonary artery remodeling. Cell Signal. 28:1489–1501. 2016. View Article : Google Scholar : PubMed/NCBI
46	Zhao H, Cui Y, Dupont J, Sun H, Hennighausen L and Yakar S: Overexpression of the tumor suppressor gene phosphatase and tensin homologue partially inhibits wnt-1-induced mammary tumorigenesis. Cancer Res. 65:6864–6873. 2005. View Article : Google Scholar : PubMed/NCBI