Cigarette smoke extract promotes proliferation of airway smooth muscle cells through suppressing C/EBP-α expression

Guan,Pin; Cai,Wentao; Yu,Huapeng; Wu,Zhiyong; Li,Wei; Wu,Jie; Chen,Juan; Feng,Guangqiu

doi:10.3892/etm.2017.4126

Cigarette smoke extract promotes proliferation of airway smooth muscle cells through suppressing C/EBP-α expression

Authors:
- Pin Guan
- Wentao Cai
- Huapeng Yu
- Zhiyong Wu
- Wei Li
- Jie Wu
- Juan Chen
- Guangqiu Feng
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Affiliations: Graduate School, Southern Medical University, Guangzhou, Guangdong 510000, P.R. China, Department of Orthopedics, People's Hospital of Hainan, Haikou, Hainan 570311, P.R. China, Department of Respiratory Medicine, Zhujiang Hospital of Southern Medical University, Guangzhou, Guangdong 510000, P.R. China, Department of Medical Center, People's Hospital of Hainan, Haikou, Hainan 570311, P.R. China, Institute of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan 570311, P.R. China
Published online on: February 15, 2017 https://doi.org/10.3892/etm.2017.4126
Pages: 1408-1414
Copyright: © Guan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cigarette smoke has been considered a major contributor to the pathogenesis of chronic obstructive pulmonary disease (COPD). In COPD patients, the airway smooth muscle layer has been observed to be markedly thickened and the proliferation of airway smooth muscle cells (ASMCs) was therefore used by the present study as a model to assess the impact of cigarette smoke extract (CSE). ASMCs were exposed to various concentrations of CSE and the proliferation of the cells was analyzed by an MTT assay. Furthermore, the expression levels of calreticulin and CCAAT/enhancer‑binding protein alpha (C/EBP-α) in CSE-stimulated ASMCs were determined by polymerase chain reaction and western blot analyses. In addition, the effects of RNA interference (RNAi) to knockdown calreticulin and/or C/EBP‑α on ASMC proliferation were studied. CSE was found to promote the proliferation of ASMCs, which was associated with increased expression of calreticulin and decreased expression of C/EBP-α. Knockdown of calreticulin resulted in the upregulation of C/EBP‑α and inhibition of cell proliferation, while simultaneous knockdown of C/EBP‑α promoted cell proliferation. The present study revealed that CSE promoted the proliferation of ASMCs, which was mediated by inhibition of C/EBP-α. These findings shed new light on airway remodeling in COPD and may provide novel approaches for therapies.

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1	Gorska K, Krenke R, Kosciuch J, Korczynski P, Zukowska M, Domagala-Kulawik J, Maskey-Warzechowska M and Chazan R: Relationship between airway inflammation and remodeling in patients with asthma and chronic obstructive pulmonary disease. Eur J Med Res. 14 Suppl 4:S90–S96. 2009.
2	Yawn BP: Differential assessment and management of asthma vs chronic obstructive pulmonary disease. Medscape J Med. 11:202009.PubMed/NCBI
3	Hogg JC, Chu F, Utokaparch S, Woods R, Elliott WM, Buzatu L, Cherniack RM, Rogers RM, Sciurba FC, Coxson HO and Paré PD: The nature of small-airway obstruction in chronic obstructive pulmonary disease. N Engl J Med. 350:2645–2653. 2004. View Article : Google Scholar : PubMed/NCBI
4	Pini L, Pinelli V, Modina D, Bezzi M, Tiberio L and Tantucci C: Central airways remodeling in COPD patients. Int J Chron Obstruct Pulmon Dis. 9:927–932. 2014. View Article : Google Scholar : PubMed/NCBI
5	Pera T, Gosens R, Lesterhuis AH, Sami R, van der Toorn M, Zaagsma J and Meurs H: Cigarette smoke and lipopolysaccharide induce a proliferative airway smooth muscle phenotype. Respir Res. 11:482010. View Article : Google Scholar : PubMed/NCBI
6	He F, Li B, Zhao Z, Zhou Y, Hu G, Zou W, Hong W, Zou Y, Jiang C, Zhao D and Ran P: The pro-proliferative effects of nicotine and its underlying mechanism on rat airway smooth muscle cells. PLoS One. 9:e935082014. View Article : Google Scholar : PubMed/NCBI
7	Zhu J, Wu YN, Zhang W, Zhang XM, Ding X, Li HQ, Geng M, Xie ZQ and Wu HM: Monocarboxylate transporter 4 facilitates cell proliferation and migration and is associated with poor prognosis in oral squamous cell carcinoma patients. PLoS One. 9:e879042014. View Article : Google Scholar : PubMed/NCBI
8	Seow CY, Schellenberg RR and Paré PD: Structural and functional changes in the airway smooth muscle of asthmatic subjects. Am J Respir Crit Care Med. 158:S179–S186. 1998. View Article : Google Scholar : PubMed/NCBI
9	Didon L, Roos AB, Elmberger GP, Gonzalez FJ and Nord M: Lung-specific inactivation of CCAAT/enhancer binding protein alpha causes a pathological pattern characteristic of COPD. Eur Respir J. 35:186–197. 2010. View Article : Google Scholar : PubMed/NCBI
10	Roth M, Johnson PR, Borger P, Bihl MP, Rüdiger JJ, King GG, Ge Q, Hostettler K, Burgess JK, Black JL and Tamm M: Dysfunctional interaction of C/EBPalpha and the glucocorticoid receptor in asthmatic bronchial smooth-muscle cells. N Engl J Med. 351:560–574. 2004. View Article : Google Scholar : PubMed/NCBI
11	Wang H, Iakova P, Wilde M, Welm A, Goode T, Roesler WJ and Timchenko NA: C/EBPalpha arrests cell proliferation through direct inhibition of Cdk2 and Cdk4. Mol Cell. 8:817–828. 2001. View Article : Google Scholar : PubMed/NCBI
12	Miglino N, Roth M, Lardinois D, Tamm M and Borger P: Calreticulin is a negative regulator of bronchial smooth muscle cell proliferation. J Allergy (Cairo). 2012:7832902012.PubMed/NCBI
13	Wang WA, Groenendyk J and Michalak M: Calreticulin signaling in health and disease. Int J Biochem Cell Biol. 44:842–846. 2012. View Article : Google Scholar : PubMed/NCBI
14	Chiang WF, Hwang TZ, Hour TC, Wang LH, Chiu CC, Chen HR, Wu YJ, Wang CC, Wang LF, Chien CY, et al: Calreticulin, an endoplasmic reticulum-resident protein, is highly expressed and essential for cell proliferation and migration in oral squamous cell carcinoma. Oral Oncol. 49:534–541. 2013. View Article : Google Scholar : PubMed/NCBI
15	Lu YC, Weng WC and Lee H: Functional roles of calreticulin in cancer biology. Biomed Res Int. 2015:5265242015. View Article : Google Scholar : PubMed/NCBI
16	Timchenko LT, Iakova P, Welm AL, Cai ZJ and Timchenko NA: Calreticulin interacts with C/EBPalpha and C/EBPbeta mRNAs and represses translation of C/EBP proteins. Mol Cell Biol. 22:7242–7257. 2002. View Article : Google Scholar : PubMed/NCBI
17	Pera T, Atmaj C, van der Vegt M, Halayko AJ, Zaagsma J and Meurs H: Role for TAK1 in cigarette smoke-induced proinflammatory signaling and IL-8 release by human airway smooth muscle cells. Am J Physiol Lung Cell Mol Physiol. 303:L272–L278. 2012. View Article : Google Scholar : PubMed/NCBI
18	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
19	Li Y, Pu G, Chen C and Yang L: Inhibition of FHL1 inhibits cigarette smoke extract-induced proliferation in pulmonary arterial smooth muscle cells. Mol Med Rep. 12:3801–3808. 2015.PubMed/NCBI
20	Mortaz E, Kraneveld AD, Smit JJ, Kool M, Lambrecht BN, Kunkel SL, Lukacs NW, Nijkamp FP and Folkerts G: Effect of cigarette smoke extract on dendritic cells and their impact on T-cell proliferation. PLoS One. 4:e49462009. View Article : Google Scholar : PubMed/NCBI
21	Wu XJ, Luo GX, Zeng X, Lan LL, Ning Q, Xu YJ, Zhao JP and Xie JG: Genotoxicity and reduced heat shock protein 70 in human airway smooth muscle cells exposed to cigarette smoke extract. J Huazhong Univ Sci Technolog Med Sci. 33:827–833. 2013. View Article : Google Scholar : PubMed/NCBI
22	Chen QW, Edvinsson L and Xu CB: Cigarette smoke extract promotes human vascular smooth muscle cell proliferation and survival through ERK1/2- and NF-κB-dependent pathways. ScientificWorldJournal. 10:2139–2156. 2010. View Article : Google Scholar : PubMed/NCBI
23	Ambalavanan N, Carlo WF, Bulger A, Shi J and Philips JB III: Effect of cigarette smoke extract on neonatal porcine vascular smooth muscle cells. Toxicol Appl Pharmacol. 170:130–136. 2001. View Article : Google Scholar : PubMed/NCBI
24	Zhang XY, Xu YJ, Liu XS and Zhang ZX: Cigarette smoke extract promotes proliferation of airway smooth muscle cells in asthmatic rats via regulating cyclin D1 expression. Chin Med J (Engl). 123:1709–1714. 2010.PubMed/NCBI
25	Helbling D, Mueller BU, Timchenko NA, Schardt J, Eyer M, Betts DR, Jotterand M, Meyer-Monard S, Fey MF and Pabst T: CBFB-SMMHC is correlated with increased calreticulin expression and suppresses the granulocytic differentiation factor CEBPA in AML with inv (16). Blood. 106:1369–1375. 2005. View Article : Google Scholar : PubMed/NCBI
26	Miglino N, Roth M, Lardinois D, Sadowski C, Tamm P and Borger P: Cigarette smoke inhibits lung fibroblast proliferation by translational mechanisms. Eur Respir J. 39:705–711. 2012. View Article : Google Scholar : PubMed/NCBI
27	Xu GN, Yang K, Xu ZP, Zhu L, Hou LN, Qi H, Chen HZ and Cui YY: Protective effects of anisodamine on cigarette smoke extract-induced airway smooth muscle cell proliferation and tracheal contractility. Toxicol Appl Pharmacol. 262:70–79. 2012. View Article : Google Scholar : PubMed/NCBI
28	Zeng DX, Xu YJ, Liu XS, Wang R and Xiang M: Cigarette smoke extract induced rat pulmonary artery smooth muscle cells proliferation via PKCα-mediated cyclin D1 expression. J Cell Biochem. 112:2082–2088. 2011. View Article : Google Scholar : PubMed/NCBI