Skip to main content
SpringerLink
Log in

Genotoxicity and reduced heat shock protein 70 in human airway smooth muscle cells exposed to cigarette smoke extract

  • Published:
Journal of Huazhong University of Science and Technology [Medical Sciences] Aims and scope Submit manuscript

Summary

Cigarette smoke is associated with the development of several diseases, such as chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate genotoxicity and heat shock protein 70 (Hsp70) in human airway smooth muscle cells (HASMCs) exposed to cigarette smoke extract (CSE). HASMCs was exposed to CSE with different doses for 24 h. The level of 8-hydroxydeoxyguanosine (8-OHdG) was determined by using HPLC-ECD, the DNA damage was analyzed by using comet assay, and apoptosis was examined by using Annexin-FITC/PI staining. The production of Hsp70 after CSE stimulation was tested. Results indicated that CSE significantly increased the level of 8-OHdG, DNA damage and cell apoptosis, and reduced the production of Hsp70. In particular, levels of Hsp70 were inversely correlated with 8-OHdG, DNA damage and cell apoptosis. It was concluded that cigarette smoke induced genotoxicity and decreased the production of cell protective protein Hsp70, which may contribute to the development of some airway diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Repine JE, Bast A, Lankhorst I. Oxidative stress in chronic obstructive pulmonary disease. Oxidative Stress Study Group. Am J Respir Crit Care Med, 1997,156(2 Pt 1):341–357

    Article  CAS  PubMed  Google Scholar 

  2. Lofroth G. Environmental tobacco smoke: overview of chemical composition and genotoxic components. Mutat Res, 1989,222(2):73–80

    Article  CAS  PubMed  Google Scholar 

  3. Pryor WA. Biological effects of cigarette smoke, wood smoke, and the smoke from plastics: the use of electron spin resonance. Free Radic Biol Med, 1992,13(6): 659–676

    Article  CAS  PubMed  Google Scholar 

  4. Agbaje IM, McVicar CM, Schock BC, et al. Increased concentrations of the oxidative DNA adduct 7,8-dihydro-8-oxo-2-deoxyguanosine in the germ-line of men with type 1 diabetes. Reprod Biomed Online, 2008,16(3):401–409

    Article  CAS  PubMed  Google Scholar 

  5. Chung KF, Adcock IM. Multifaceted mechanisms in COPD: inflammation, immunity, and tissue repair and destruction. Eur Respir J, 2008,31(6):1334–1356

    Article  CAS  PubMed  Google Scholar 

  6. Vayssier M, Banzet N, Francois D, et al. Tobacco smoke induces both apoptosis and necrosis in mammalian cells: differential effects of HSP70. Am J Physiol, 1998,275(4 Pt 1):L771–779

    CAS  PubMed  Google Scholar 

  7. Hightower LE. Heat shock, stress proteins, chaperones, and proteotoxicity. Cell, 1991,66(2):191–197

    Article  CAS  PubMed  Google Scholar 

  8. Wu T, Chen S, Xiao C, et al. Presence of antibody against the inducible Hsp71 in patients with acute heat-induced illness. Cell Stress Chaperones, 2001,6(2): 113–120

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Schroder HC, Hassanein HM, Lauenroth S, et al. Induction of DNA strand breaks and expression of HSP70 and GRP78 homolog by cadmium in the marine sponge Suberites domuncula. Arch Environ Contam Toxicol, 1999,36(1):47–55

    Article  CAS  PubMed  Google Scholar 

  10. Zhang K, Shan L, Rahman MS, et al. Constitutive and inducible thymic stromal lymphopoietin expression in human airway smooth muscle cells: role in chronic obstructive pulmonary disease. Am J Physiol Lung Cell Mol Physiol, 2007,293(2):L375–382

    Article  CAS  PubMed  Google Scholar 

  11. Twort C, Van Breemen C. Human airway smooth muscle in culture. Tissue Cell, 1988,20(3):339–344

    Article  CAS  PubMed  Google Scholar 

  12. Lee SD, Lee DS, Chun YG, et al. Cigarette smoke extract induces endothelin-1 via protein kinase C in pulmonary artery endothelial cells. Am J Physiol Lung Cell Mol Physiol, 2001,281(2):L403–411

    CAS  PubMed  Google Scholar 

  13. Helbock HJ, Beckman KB, Shigenaga MK, et al. DNA oxidation matters: the HPLC-electrochemical detection assay of 8-oxo-deoxyguanosine and 8-oxo-guanine. Proc Natl Acad Sci USA, 1998,95(1):288–293

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. Pouget JP, Douki T, Richard MJ, et al. DNA damage induced in cells by gamma and UVA radiation as measured by HPLC/GC-MS and HPLC-EC and Comet assay. Chem Res Toxicol, 2000,13(7):541–549

    Article  CAS  PubMed  Google Scholar 

  15. Kindzelskii AL, Petty HR. Ultrasensitive detection of hydrogen peroxide-mediated DNA damage after alkaline single cell gel electrophoresis using occultation microscopy and TUNEL labeling. Mutat Res, 1999,426(1): 11–22

    Article  CAS  PubMed  Google Scholar 

  16. Hodge GL, Flower R, Han P. Optimal storage conditions for preserving granulocyte viability as monitored by Annexin V binding in whole blood. J Immunol Methods, 1999,225(1-2):27–38

    Article  CAS  PubMed  Google Scholar 

  17. Wu T, Yuan Y, Wu Y, et al. Presence of antibodies to heat stress proteins in workers exposed to benzene and in patients with benzene poisoning. Cell Stress Chaperones, 1998,3(3):161–167

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  18. Carnevali S, Petruzzelli S, Longoni B, et al. Cigarette smoke extract induces oxidative stress and apoptosis in human lung fibroblasts. Am J Physiol Lung Cell Mol Physiol, 2003,284(6):L955–963

    CAS  PubMed  Google Scholar 

  19. Liu X, Conner H, Kobayashi T, et al. Cigarette smoke extract induces DNA damage but not apoptosis in human bronchial epithelial cells. Am J Respir Cell Mol Biol, 2005,33(2):121–129

    Article  PubMed  Google Scholar 

  20. Shenberger JS, Dixon PS. Oxygen induces S-phase growth arrest and increases p53 and p21(WAF1/CIP1) expression in human bronchial smooth-muscle cells. Am J Respir Cell Mol Biol, 1999,21(3):395–402

    Article  CAS  PubMed  Google Scholar 

  21. Pytel D, Wysocki T, Majsterek I. Comparative study of DNA damage, cell cycle and apoptosis in human K562 and CCRF-CEM leukemia cells: Role of BCR/ABL in therapeutic resistance. Comp Biochem Physiol C Toxicol Pharmacol, 2006,144(1):85–92

    Article  PubMed  Google Scholar 

  22. Cheetham ME, Anderton BH, Jackson AP. Inhibition of hsc70-catalysed clathrin uncoating by HSJ1 proteins. Biochem J, 1996,319 ( Pt 1):103–108

    Google Scholar 

  23. Hartl FU. Molecular chaperones in cellular protein folding. Nature, 1996,381(6583):571–579

    Article  CAS  PubMed  Google Scholar 

  24. Vayssier-Taussat M, Camilli T, Aron Y, et al. Effects of tobacco smoke and benzo[a]pyrene on human endothelial cell and monocyte stress responses. Am J Physiol Heart Circ Physiol, 2001,280(3):H1293–1300

    CAS  PubMed  Google Scholar 

  25. Gao YJ, Xiao CF, Chen S, et al. In vitro study on role of Hsp70 expression in DNA damage of human embryonic lung cells exposed to Benzo[a]pyrene. Biomed Environ Sci, 2004,17(2):144–152

    PubMed  Google Scholar 

  26. Chen YC, Lin-Shiau SY. Involvement of heat-shock protein 70 and P53 proteins in attenuation of UVC-induced apoptosis by thermal stress in hepatocellular carcinoma cells. Photochem Photobiol, 1999,70(1): 78–86

    Article  CAS  PubMed  Google Scholar 

  27. Calini V, Urani C, Camatini M. Overexpression of HSP70 is induced by ionizing radiation in C3H 10T1/2 cells and protects from DNA damage. Toxicol In Vitro, 2003,17(5–6):561–566

    Article  CAS  PubMed  Google Scholar 

  28. Li S, Zheng J, Carmichael ST. Increased oxidative protein and DNA damage but decreased stress response in the aged brain following experimental stroke. Neurobiol Dis, 2005,18(3):432–440

    Article  CAS  PubMed  Google Scholar 

  29. Hamzeh M, Sunahara GI. In vitro cytotoxicity and genotoxicity studies of titanium dioxide (TiO(2)) nanoparticles in Chinese hamster lung fibroblast cells. Toxicol In Vitro, 2012,27(2):864–873

    Article  PubMed  Google Scholar 

  30. Tennen RI, Haye JE, Wijayatilake HD, et al. Cell-cycle and DNA damage regulation of the DNA mismatch repair protein Msh2 occurs at the transcriptional and post-transcriptional level. DNA Repair (Amst), 2013,12(2):97–109

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Respiratory Medicine and Critical Care Medicine, Huazhong University of Science and Technology, Wuhan, 430030, China

    Xiao-jie Wu  (武小杰), Guo-xiong Luo  (罗国雄), Xue Zeng  (曾 雪), Li-li Lan  (兰立立), Yong-jian Xu  (徐永健), Jian-ping Zhao  (赵建平) & Jun-gang Xie  (谢俊刚)

  2. Department of Infectious Disease, Institute of Infectious Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Qin Ning  (宁 琴)

Authors
  1. Xiao-jie Wu  (武小杰)
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Guo-xiong Luo  (罗国雄)
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xue Zeng  (曾 雪)
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Li-li Lan  (兰立立)
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qin Ning  (宁 琴)
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yong-jian Xu  (徐永健)
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jian-ping Zhao  (赵建平)
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jun-gang Xie  (谢俊刚)
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun-gang Xie  (谢俊刚).

Additional information

This project was supported by grants from the National Natural Science Foundation of China (No.81070036, 81370145, 81370156 and 81070021), The National Key Technology R&D Program of the 12th National Five-year Development Plan: Clinical Study on Translational Medicine of Respiratory Disease (No. 212BA105B01), The Specific Project of National Health Research Project of Ministry of Health of China (No. 201002008) and Program for Changjiang Scholars and Innovative Research Team in University (No. PCSIRT1131).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wu, Xj., Luo, Gx., Zeng, X. et al. Genotoxicity and reduced heat shock protein 70 in human airway smooth muscle cells exposed to cigarette smoke extract. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 33, 827–833 (2013). https://doi.org/10.1007/s11596-013-1206-8

Download citation

  • Received:

  • Revised:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11596-013-1206-8

Key words

Search

Navigation