Abstract
Lung disease is the leading and second-leading cause of death in women and men in Taiwan, respectively. Epidemiological studies conducted in Taiwan have shown that cigarette smoking is the principal risk factor of lung disease, but little is known about the association between apoptosis and cigarette smoke (CS)-induced lung pathogenesis. We designed an animal exposure system to study signal proteins involved in the process of apoptosis induced by smoking in rat terminal bronchiole. Rats were exposed to CS in doses of 5, 10, and 15 cigarettes, respectively, and the exposure lasted for 30 min, twice a day, 6 days a week for 1 month. Following which the rats were sacrificed and the lung tissues were analyzed by histopathological methods. The terminal bronchioles revealed mild to severe inflammation according to the doses of CS and marked lipid peroxidation, lymphocyte infiltration, congestion, and epithelial emphysema of alveolar spaces were also noted. Using an in situ cell death detection kit (TA300), the association of CS with apoptosis was determined in a concentration-dependent manner. Immunohistochemical evaluation showed that CS treatment produced an increase in the cellular levels of Bax, t-Bid, cleaved caspase-3, phospho-p53, phospho-JNK, and FasL but a decline in Bcl-2 and Mcl-1 (p<0.001 for all) in rat terminal bronchioles. The results provided evidences suggesting that exposure to CS not only induced apoptosis, but also involved p53/Bax and JNK/FasL cascade pathway.
Similar content being viewed by others
Abbreviations
- CS:
-
Cigarette smoke
- Bax:
-
Bcl-2 associated X protein
- t-Bid:
-
Truncated Bid
- Bcl-2:
-
B cell lymphoma-2
- MAPK:
-
Mitogen-activated protein kinases family
- JNK:
-
c-Jun N-terminal kinase
- Fas-L:
-
Fas ligand
- DAB:
-
Diaminobenzidine
- iNOS:
-
Inducible NO synthase
- NOx:
-
Nitrogen oxides
- HPF:
-
High-power-field
References
Adams JM, Cory S (1998) The Bcl-2 protein family: arbiters of cell survival. Science 281:1322–1326
Alnemri ES (1997) Mammalian cell death proteases: a family of highly conserved aspartate specific cysteine proteases. J Cell Biochem 64:33–42
Asami S, Hirano T, Yamaguchi R, Tomioka Y, Itoh H, Kasai H (1996) Increase of a type of oxidative DNA damage, 8-hydroxyguanine and its repair activity in human leukocytes by cigarette smoking. Cancer Res 56:2546–2549
Asami S, Manabe H, Miyake J, Tsurudome Y, Hirano T, Yamaguchi R, Itoh H, Kasai H (1997) Cigarette smoking induces an increase in oxidative DNA damage, 8-hydroxydeoxyguanosine, in a central site of the human lung. Carcinogenesis 18:1763–1766
Butterfield L, Storey B, Maas L, Heasley LE (1997) c-Jun NH2-terminal kinase regulation of the apoptotic response of small cell lung cancer cells to ultraviolet radiation. J Biol Chem 272:10110–10116
Chang WC, Lee YC, Liu CL, Hsu JD, Wang HC, Chen CC, Wang CJ (2001) Increased expression of iNOS and c-fos via regulation of protein tyrosine phosphorylation and MEK1/ERK2 proteins in terminal bronchiole lesions in the lungs of rats exposed to cigarette smoke. Arch Toxicol 75:28–35
Chan-Hui PY, Weaver R (1998) Human mitogen-activated protein kinase kinase kinase mediates the stress-induced activation of mitogen-activated protein kinase cascades. Biochem J 336:599–609
Cobb MH, Goldsmith EJ (1995) How MAP kinases are regulated. J Biol Chem 270:14843–14846
Czekaj P, Palasz A, Lebda-Wyborny T, Nowaczyk-Dura G, Karczewska W, Florek E, Kaminski M (2002) Morphological changes in lungs, placenta, liver and kidneys of pregnant rats exposed to cigarette smoke. Int Arch Occup Environ Health 75(Suppl):S27–S35
Danial NN, Korsmeyer SJ (2004) Cell death: critical control points. Cell 116:205–219
De Flora S, D’Agostini F, Balansky R, Camoirano A, Bennicelli C, Bagnasco M, Cartiglia C, Tampa E, Longobardi MG, Lubet RA, Izzotti A (2003) Modulation of cigarette smoke-related end-points in mutagenesis and carcinogenesis. Mutat Res 523–524:237–252
Fielding S, Short C, Davies K, Wald N, Bridges BA, Waters R (1989) Studies on the ability of smoke from different types of cigarettes to induce DNA single-strand breaks in cultured human cells. Mutat Res 214:147–151
Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312
Greenlee RT, Hill-Harmon MB, Murray T, Thun M (2001) Cancer statistics, CA. Cancer J Clin 51:15–36
Herr I, Debatin KM (2001) Cellular stress response and apoptosis in cancer therapy. Blood 98:2603–2614
Hoshino S, Yoshida M, Inoue K, Yano Y, Yanagita M, Mawatari H, Yamane H, Kijima T, Kumagai T, Osaki T, Tachiba I, Kawase I (2005) Cigarette smoke extract induces endothelial cell injury via JNK pathway. Biochem Biophys Res Commun 329:58–63
Hsieh YS, Wang HC, Tseng TW, Chang WC, Wang CJ (2001) Gaseous nitric oxide-induced 8-nitroguanine formation in human lung fibroblast cells and cell-free DNA. Toxicol Appl Pharmacol 172:210–216
Hsieh YS, Chen BC, Shiow SJ, Wang HC, Hsu JD, Wang CJ (2002) Formation of 8-nitroguanine in tobacco cigarette smokers and in tobacco smoke-exposed Wistar rats. Chem Biol Interact 140:67–80
Ichijo H (1999) From receptors to stress-activated MAP kinases. Oncogene 18:6087–6093
Jacobson MD, Raff MC (1995) Programmed cell death and Bcl-2 protection in very low oxygen. Nature 374:814–816
Juo P, Kuo CJ, Reynolds SE, Konz RF, Raingeaud J, Davis RJ, Biemann HP, Blenis J (1997) Fas activation of the p38 mitogen-activated protein kinase signalling pathway requires ICE/CED-3 family proteases. Mol Cell Biol 17:24–35
Ko YC, Lee CH, Chen MJ, Huang CC, Chang WY, Lin HJ, Wang HZ, Chang PY (1997) Risk factors for primary lung cancer among non-smoking women in Taiwan. Int J Epidemiol 26:24–31
Leanderson P, Tagesson C (1992) Cigarette smoke-induced DNA damage in cultured human lung cells: role of hydroxy radicals and endonuclease activation. Chem Biol Interact 81:197–208
Lei W, Yu R, Mandlekar S, Kong AN (1998) Induction of apoptosis and activation of interleukin 1beta-converting enzyme/Ced-3 protease (caspase-3) and c-Jun NH2-terminal kinase 1 by benzo(a)pyrene. Cancer Res 58:2102–2106
Li H, Zhu H, Xu CJ, Yuan J (1998) Cleavage of Bid by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 94:491–501
Lowe SW, Lin AW (2000) Apoptosis in cancer. Carcinogenesis 21:485–495
Luo X, Budihardjo I, Zou H, Slaughter C, Wang X (1998) Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 94:481–490
Muller T, Gebel S (1994) Heme oxygenase expression in Swiss 3T3 cells following exposure to aqueous cigarette smoke fractions. Carcinogenesis 15:67–72
Muller T (1995)Expression of c-fos in quiescent Swiss 3T3 cells exposed to aqueous cigarette smoke fractions. Cancer Res 55:1927–1932
Muller T, Haussmann HJ, Schepers G (1997) Evidence for peroxynitrite as an oxidative stress-inducing compound of aqueous cigarette smoke fractions. Carcinogenesis 18:295–301
Nakayama T, Kaneko M, Kodama M, Nagata C (1984) Cigarette smoke induced DNA single-strand breaks in human cells. Nature 314:462–464
Oltvai ZN, Milliman CL, Korsmeyer SJ (1993) Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74:609–619
Raveendran M, Wang J, Senthil D, Wang J, Utama B, Shen Y, Dudley D, Zhang Y, Wang XL (2005) Endogenous nitric oxide activation protects against cigarette smoking induced apoptosis in endothelial cells. FEBS Lett 579:733–740
Roulston A, Reinhard C, Amiri P, Williams LT (1998) Early activation of c-Jun N-terminal kinase and p38 kinase regulate cell survival in response to tumor necrosis factor alpha. J Biol Chem 273:10232–10239
Seimiya H, Mashima T, Toho M, Tsuruo T (1997) c-Jun NH2-terminal kinase-mediated activation of interleukin-1beta converting enzyme/CED-3-like protease during anticancer drug-induced apoptosis. J Biol Chem 272:4631–4636
Shishodia S, Aggarwal BB (2004) Cyclooxygenase (COX)-2 inhibitor celecoxib abrogates activation of cigarette smoke-induced nuclear factor (NF)-kappaB by suppressing activation of IkappaBalpha kinase in human non-small cell lung carcinoma: correlation with suppression of cyclin D1, COX-2, and matrix metalloproteinase-9. Cancer Res 64:5004–5012
Spencer JP, Jenner A, Chimel K, Ardoma OL, Cross CE, Wu R, Halliwell B (1995) DNA damage in human respiratory tract epithelial cells: damage by gas phase cigarette smoke apparently involves attack by reactive nitrogen species in addition to oxygen radicals. FEBS Lett 375:179–182
Stone KK, Bermudez E, Pryor WA (1994) Aqueous extracts of cigarette tar containing the tar free radical cause DNA nicks in mammalian cells. Environ Health Perspect (Suppl)102:173–178
Stone K, Bermudez E, Zang LY, Carter KM, Queenan KE, Pryor WA (1995) The ESR properties, DNA nicking, and DNA association of aged solutions of catechol versus aqueous extracts of tar from cigarette smoke. Arch Biochem Biophys 319:196–203
Tewari M, Quan LT, O’Rourke K, Desnoyers S, Zeng Z, Beidler DR, Poirier GG, Salvesen GS, Dixit VM (1995) Yama/CPP32 beta, a mammalian homolog for CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell 81:801–809
Thompson CB (1995) Apoptosis in the pathogenesis and treatment of disease. Science 226:1097–1099
Villeneuve PJ, Morrison HI (1995) Trends in mortality from smoking related cancers, 1950 to 1991. Can Soc Trends 39:8–11
Vineis P, Caporaso N (1995) Tobacco and cancer epidemiology and the laboratory. Environ Health Perspect 103:156–160
Yermilov V, Rubio J, Becchi M, Friesen MD, Pignatelli B, Ohshima H (1995a) Formation of 8-nitroguanine by the reaction of guanine with peroxynitrite in vitro. Carcinogenesis 16:2045–2050
Yermilov V, Rubio J, Ohshima H (1995b) Formation of 8-nitroguanine in DNA treated with peroxynitrite in vitro and its rapid removal from DNA by depurination. FEBS Lett 376:207–210
Yin XM (2000) Signal transduction mediated by Bid, a pro-death Bcl-2 family protein, connects the death receptor and mitochondria apoptosis pathways. Cell Res 10:161–167
Acknowledgment
This work was supported by a grant from the National Science Council (NSC 94-2320-B040-039), Taiwan.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wu, CH., Lin, HH., Yan, FP. et al. Immunohistochemical Detection of Apoptotic Proteins, p53/Bax and JNK/FasL Cascade, in the Lung of Rats Exposed to Cigarette Smoke. Arch Toxicol 80, 328–336 (2006). https://doi.org/10.1007/s00204-005-0050-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-005-0050-4