Induction of apoptosis with tobacco smoke and related products in A549 lung epithelial cells in vitro

被引：56

作者：

Ramage L. ^{[1
]}

Jones A.C. ^{[1
]}

Whelan C.J. ^{[1
]}

机构：

[1] School of Life Sciences, University of Hertfordshire, Hatfield, Herts., AL10 9AB, College Lane

来源：

Journal of Inflammation | / 3卷 / 1期

关键词：

Nicotine; A549 Cell; Tobacco Smoke; Taxol; Airway Epithelial Cell;

D O I：

10.1186/1476-9255-3-3

中图分类号：

学科分类号：

摘要：

Background: This study has investigated the ability of tobacco smoke, and ingredients of tobacco smoke, to induce apoptosis in the airway epithelial cell line A549. Method: A549 cells were treated with 80 μg/ml Tobacco smoke condensate (TSC), 10 mM Nicotine, 10 μM paraldehyde, 10 μM hydrogen peroxide, 1 μM Taxol® (Paclitaxel), 100%, 50% and 25% cigarette smoke extract (CSE). Following 4-48 h incubation apoptosis was measured morphologically following staining of cells with DAPI. TUNEL staining was also used to assess DNA damage after 24 and 48 h incubation. In addition, loss of mitochondrial cytochrome C and activation of Bax-α, early events in the apoptotic process, were measured after 4 h of incubation. Results: Incubation of A549 cells with vehicle, Taxol, TSC, nicotine, paraldehyde, hydrogen peroxide and CSE caused a time-dependent detachment of the cells from the flask between 6 and 48 h. DAPI staining revealed that the cells remaining adhered to the flask appeared healthy whereas some of those that had detached appeared to be either apoptotic or indeterminate. Treatment with Taxol, TSC, nicotine, paraldehyde, hydrogen peroxide and CSE caused a significant increase in the number of apoptotic cells. Similarly, treatment with Taxol, TSC, nicotine, hydrogen peroxide and CSE caused a significant increase in the number of apoptotic cells among the cells that had detached from the culture plate. After 4 h of incubation, Taxol, TSC, hydrogen peroxide and CSE caused a significant reduction in mitochondrial cytochrome C and an increase in cytosolic cytochrome C. At the same time point, hydrogen peroxide and CSE significantly increased the concentration of Bax-α in the mitochondria. Conclusion: Tobacco smoke initiates apoptosis in A549 airway epithelial cells as a result of mitochondrial damage and that this results in a cell detachment and full apoptosis. This effect appears to result from factors in tobacco smoke other than nicotine and may result from free radical activity. However, additional stable factors may also be involved since the free radical content of TSC is likely to be low. © 2006Ramage et al; licensee BioMed Central Ltd.

引用

共 18 条

[1]

MacNee W., Oxidants, antioxidants and COPD, Chest, 117, (2000)

[2]

Spurzem J.R., Rennard S.I., Pathogenesis of COPD, Semin Respir Crit Care Med, 26, pp. 142-153, (2005)

[3]

Church D.F., Pryor W.A., Free-radical chemistry of cigarette smoke and toxicological implications, Environ Health Perspec, 64, pp. 111-126, (1985)

[4]

Pryor W.A., The free radical chemistry of cigarette smoke and their interaction of alpha-1-proteinase inhibitor, Pulmonary Emphysema and Proteolysis, pp. 369-393, (1986)

[5]

Hoshino Y., Mio T., Nagai S., Miki H., Ito I., Izumi T., Cytotoxic effects of cigarette smoke extract on an alveolar type II cell-derived cell line, Am J Physiol Lung Cell Mol Physiol, 281, (2001)

[6]

Lannan S., Donaldson K., Brown D., MacNee W., Effect of cigarette smoke and its condensates on alveolar epithelial cell injury in vitro, Am J Physiol, 266, (1994)

[7]

Kim H., Liu X., Kobayashi T., Conner H., Kohyama T., Wen F.Q., Fang Q., Abe S., Bitterman P., Rennard S.I., Reversible cigarette smoke extract-induced DNA damage in human lung fibroblasts, Am J Respir Cell Mol Biol, 31, pp. 483-490, (2004)

[8]

Yokohori N., Aoshiba K., Nagai A., Increased levels of cell death and proliferation in alveolar wall cells in patients with pulmonary emphysema, Chest, 125, pp. 626-632, (2004)

[9]

Wickenden J.A., Clarke M.C., Rossi A.G., Rahman I., Faux S.P., Donaldson K., MacNee W., Cigarette smoke prevents apoptosis through inhibition of caspase activation and induces necrosis, Am J Respir Cell Mol Biol, 29, pp. 562-570, (2003)

[10]

Kerr J.F., Wyllie A.H., Currie A.R., Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics, Br J Cancer, 26, pp. 239-257, (1972)

← 1 2 →