Abstract
Smoking tobacco is a major risk factor for the development of lung cancer, COPD, and other lung pathologies in smokers. Cigarette smoke (CS), which is comprised of several toxic components, is known to cause oxidative stress and inflammation-induced lung damage. Since airway epithelial cells act as the primary barrier, they protect the lung tissues from environmental insults, including CS. Upon exposure to these insults, airway epithelial cells act as the initial site of injury and orchestrate the pathophysiology of lung cancer. Scientists have been using cigarette smoke extract (CSE) in the preclinical model of in vitro cell culture to understand the effect of CS on the cellular, biochemical, and molecular changes in the lung epithelial cells. However, the standard procedure to prepare the CSE in the laboratory with a low-cost assembly and obtaining a reproducible quality of CSE in different batches is a challenge. Here, in this chapter, we delineate the method for the preparation of CSE using a discontinuous puff-based system which is an economical and reproducible method to prepare CSE in the laboratory. This method is suitable for studying CSE-induced molecular changes in lung diseases, including lung cancer, using in vitro models of lung adenocarcinoma cells.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lira-Junior R, Åkerman S, Gustafsson A, Klinge B, Boström EA (2017) Colony stimulating factor-1 in saliva in relation to age, smoking, and oral and systemic diseases. Sci Rep 7(1):7280. https://doi.org/10.1038/s41598-017-07698-4
Zhang Y-J, Zhang L, Huang X-L, Duan Y, Yang L-J, Wang J (2017) Association between cigarette smoking and impaired clinical symptoms in systemic sclerosis: a review. Cell Immunol 318:1–7. https://doi.org/10.1016/j.cellimm.2017.04.002
Raval RD, Sharma P, Chandran S, Vasavada D, Nadig P, Bakutra G (2017) To evaluate and compare periodontal disease and smoking as a parallel risk factor for systemic health by gauging the serum C-reactive protein levels. J Clin Diagn Res 11(3):ZC79–ZC82. https://doi.org/10.7860/JCDR/2017/22778.9599
O’Keeffe LM, Taylor G, Huxley RR, Mitchell P, Woodward M, Peters SAE (2018) Smoking as a risk factor for lung cancer in women and men: a systematic review and meta-analysis. BMJ Open 8(10):e021611. https://doi.org/10.1136/bmjopen-2018-021611
Hoshino Y, Mio T, Nagai S, Miki H, Ito I, Izumi T (2001) Cytotoxic effects of cigarette smoke extract on an alveolar type II cell-derived cell line. Am J Phys Lung Cell Mol Phys 281(2):L509–L516. https://doi.org/10.1152/ajplung.2001.281.2.L509
Zeng N, Wang T, Chen M, Yuan Z, Qin J, Wu Y, Gao L, Shen Y, Chen L, Wen F (2019) Cigarette smoke extract alters genome-wide profiles of circular RNAs and mRNAs in primary human small airway epithelial cells. J Cell Mol Med 23(8):5532–5541. https://doi.org/10.1111/jcmm.14436
He B, Chen Q, Zhou D, Wang L, Liu Z (2019) Role of reciprocal interaction between autophagy and endoplasmic reticulum stress in apoptosis of human bronchial epithelial cells induced by cigarette smoke extract. IUBMB Life 71(1):66–80. https://doi.org/10.1002/iub.1937
Alshehri M, Brand O, Alqarni A, Pasini A, Pang L (2017) P115Â Cigarettes smoke extract induces inflammatory gene expression in human bronchial epithelial cells. Thorax 72(Suppl 3):A146. https://doi.org/10.1136/thoraxjnl-2017-210983.257
Costello MR, Reynaga DD, Mojica CY, Zaveri NT, Belluzzi JD, Leslie FM (2014) Comparison of the reinforcing properties of nicotine and cigarette smoke extract in rats. Neuropsychopharmacology 39(8):1843–1851. https://doi.org/10.1038/npp.2014.31
Arnold MM, Loughlin SE, Belluzzi JD, Leslie FM (2014) Reinforcing and neural activating effects of norharmane, a non-nicotine tobacco constituent, alone and in combination with nicotine. Neuropharmacology 85:293–304. https://doi.org/10.1016/j.neuropharm.2014.05.035
Uchiyama S, Hayashida H, Izu R, Inaba Y, Nakagome H, Kunugita N (2015) Determination of nicotine, tar, volatile organic compounds and carbonyls in mainstream cigarette smoke using a glass filter and a sorbent cartridge followed by the two-phase/one-pot elution method with carbon disulfide and methanol. J Chromatogr A 1426:48–55. https://doi.org/10.1016/j.chroma.2015.11.058
Jamalzadeh L, Ghafoori H, Sariri R, Rabuti H, Nasirzade J, Hasani H, Aghamaali MR (2016) Cytotoxic effects of some common organic solvents on MCF-7, RAW-264.7 and human umbilical vein endothelial cells. Avicenna J Med Biochem 4(1):10–33453. https://doi.org/10.17795/ajmb-33453
Adamson J, Azzopardi D, Errington G, Dickens C, McAughey J, Gaça MD (2011) Assessment of an in vitro whole cigarette smoke exposure system: the Borgwaldt RM20S 8-syringe smoking machine. Chem Cent J 5(1):50. https://doi.org/10.1186/1752-153X-5-50
Agraval H, Yadav UCS (2019) MMP-2 and MMP-9 mediate cigarette smoke extract-induced epithelial-mesenchymal transition in airway epithelial cells via EGFR/Akt/GSK3β/β-catenin pathway: amelioration by fisetin. Chem Biol Interact 314:108846. https://doi.org/10.1016/j.cbi.2019.108846
Luppi F, Aarbiou J, van Wetering S, Rahman I, de Boer WI, Rabe KF, Hiemstra PS (2005) Effects of cigarette smoke condensate on proliferation and wound closure of bronchial epithelial cells in vitro: role of glutathione. Respir Res 6(1):140. https://doi.org/10.1186/1465-9921-6-140
Igari K, Kelly MJ, Yamanouchi D (2019) Cigarette smoke extract activates tartrate-resistant acid phosphatase-positive macrophage. J Vasc Res 56(3):139–151. https://doi.org/10.1159/000498893
Xing AP, Du YC, Hu XY, Xu JY, Zhang HP, Li Y, Nie X (2012) Cigarette smoke extract stimulates rat pulmonary artery smooth muscle cell proliferation via PKC-PDGFB signaling. J Biomed Biotechnol 2012:534384. https://doi.org/10.1155/2012/534384
Acknowledgments
HA acknowledges the Department of Science and Technology (DST), Government of India for INSPIRE fellowship. JS acknowledges Junior Research Fellowship from the University Grant Commission, New Delhi, India.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Agraval, H., Sharma, J.R., Yadav, U.C.S. (2022). Method of Preparation of Cigarette Smoke Extract to Assess Lung Cancer-Associated Changes in Airway Epithelial Cells. In: Deep, G. (eds) Cancer Biomarkers. Methods in Molecular Biology, vol 2413. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1896-7_13
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1896-7_13
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1895-0
Online ISBN: 978-1-0716-1896-7
eBook Packages: Springer Protocols