바로가기메뉴

본문 바로가기 주메뉴 바로가기

Tuberculosis & Respiratory Diseases

Article Contents

Prev Next
e-Submission

Vol.71 No.2
Citation Share

Inhibition of PKC Epsilon Attenuates Cigarette Smoke Extract- Induced Apoptosis in Human Lung Fibroblasts (MRC-5 Cells)

Tuberculosis & Respiratory Diseases / Tuberculosis & Respiratory Diseases,
2011, v.71 no.2, pp.88-96
Shin Myung Kang (Gachon University)

Jin Young Yoon (Gachon University)
Yu Jin Kim (Gachon University Gil Hospital)


Shin, M. K. , Jin, Y. Y. , Yu, J. K. , & (2011). Inhibition of PKC Epsilon Attenuates Cigarette Smoke Extract- Induced Apoptosis in Human Lung Fibroblasts (MRC-5 Cells). Tuberculosis & Respiratory Diseases, 71(2), 88-96.
  • Downloaded
  • Viewed
PDF Download

Abstract

Background: It is known that cigarette smoke (CS) causes cell death. Apoptotic cell death is involved in the pathogenesis of CS-related lung diseases. Some members of the protein kinase C (PKC) family have roles in cigarette smoke extract (CSE)-induced apoptosis. This study was conducted to investigate the role of PKC epsilon in CSE-induced apoptosis in human lung fibroblast cell line, MRC-5. Methods: Lactate dehydrogenase release was measured using a cytotoxicity detection kit. The MTT assay was used to measure cell viability. Western immunoblot, Hoechst 33342 staining and flow cytometry were used to demonstrate the effect of PKCε. Caspase-3 and caspase-8 activities were determined using a colorimetric assay. To examine PKCε activation, Western blotting was performed using both fractions of membrane and cytosol. Results: We showed that CSE activated PKCε by demonstrating increased expression of PKCε in the plasma membrane fraction. Pre-treatment of PKCε peptide inhibitor attenuated CSE-induced apoptotic cell death, as demonstrated by the MTT assay (13.03% of control, 85.66% of CSE-treatment, and 53.73% of PKCε peptide inhibitor-pre-treatment, respectively), Hoechst 33342 staining, and flow cytometry (85.64% of CSE-treatment, 53.73% of PKCε peptide inhibitor-pre-treatment). Pre-treatment of PKCε peptide inhibitor reduced caspase-3 expression and attenuated caspase-3, caspase-8 activity compared with CSE treatment alone. Conclusion: PKCε seem to have pro-apoptotic function and exerts its function through the extrinsic apoptotic pathway in CSE-exposed MRC-5 cells. This study suggests that PKCε inhibition may be a therapeutic strategy in CS-related lung disease such as chronic obstructive pulmonary disease.

keywords
Apoptosis, Cigarette Smoking, Protein Kinase C-epsilon

Reference

1.

1. Sakao S, Tatsumi K, Hashimoto T, Igari H, Shino Y, Shirasawa H, et al. Vascular endothelial growth factor and the risk of smoking-related COPD. Chest 2003;124: 323-7.

2.

2. Sugano N, Ito K. Nicotine switches the form of H2O2- induced cell death from apoptosis to necrosis in U937 cells. Immunol Lett 2000;72:163-6.

3.

3. Wang J, Wilcken DE, Wang XL. Cigarette smoke activates caspase-3 to induce apoptosis of human umbilical venous endothelial cells. Mol Genet Metab 2001;72:82-8.

4.

4. Aoshiba K, Tamaoki J, Nagai A. Acute cigarette smoke exposure induces apoptosis of alveolar macrophages. Am J Physiol Lung Cell Mol Physiol 2001;281:L1392- 401.

5.

5. Kuo WH, Chen JH, Lin HH, Chen BC, Hsu JD, Wang CJ. Induction of apoptosis in the lung tissue from rats exposed to cigarette smoke involves p38/JNK MAPK pathway. Chem Biol Interact 2005;155:31-42.

6.

6. Wu CH, Lin HH, Yan FP, Wu CH, Wang CJ. Immunohistochemical detection of apoptotic proteins, p53/Bax and JNK/FasL cascade, in the lung of rats exposed to cigarette smoke. Arch Toxicol 2006;80:328-36.

7.

7. Wang X, Zhao J, Tang S, Lee S, Glazer RI, Hewlett I. c-FLIPL regulates PKC via AP-2 to inhibit Bax-mediated apoptosis induced by HIV-1 gp120 in Jurkat cells. Mol Cell Biochem 2009;330:23-9.

8.

8. Chen JL, Lin HH, Kim KJ, Lin A, Ou JH, Ann DK. PKC delta signaling: a dual role in regulating hypoxic stress-induced autophagy and apoptosis. Autophagy 2009;5:244-6.

9.

9. Li X, Pabla N, Wei Q, Dong G, Messing RO, Wang CY, et al. PKC-delta promotes renal tubular cell apoptosis associated with proteinuria. J Am Soc Nephrol 2010; 21:1115-24.

10.

10. Park JW, Kim HP, Lee SJ, Wang X, Wang Y, Ifedigbo E, et al. Protein kinase C alpha and zeta differentially regulate death-inducing signaling complex formation in cigarette smoke extract-induced apoptosis. J Immunol 2008;180:4668-78.

11.

11. Kim H, Liu X, Kobayashi T, Conner H, Kohyama T, Wen FQ, et al. Reversible cigarette smoke extract-induced DNA damage in human lung fibroblasts. Am J Respir Cell Mol Biol 2004;31:483-90.

12.

12. Liu X, Conner H, Kobayashi T, Kim H, Wen F, Abe S, et al. Cigarette smoke extract induces DNA damage but not apoptosis in human bronchial epithelial cells. Am J Respir Cell Mol Biol 2005;33:121-9.

13.

13. Maher P. How protein kinase C activation protects nerve cells from oxidative stress-induced cell death. J Neurosci 2001;21:2929-38.

14.

14. Reshef A, Capua ND, Sperling O, Zoref-Shani E. Ischemic tolerance conferred to cultured rat neurons by heat shock is not mediated by opening of adenosine triphosphate-sensitive potassium channels. Neurosci Lett 2000;287:223-6.

15.

15. Shizukuda Y, Buttrick PM. Protein kinase C(epsilon) modulates apoptosis induced by beta -adrenergic stimulation in adult rat ventricular myocytes via extracellular signal-regulated kinase (ERK) activity. J Mol Cell Cardiol 2001;33:1791-803.

16.

16. Lee YJ, Soh JW, Jeoung DI, Cho CK, Jhon GJ, Lee SJ, et al. PKC epsilon -mediated ERK1/2 activation involved in radiation-induced cell death in NIH3T3 cells. Biochim Biophys Acta 2003;1593:219-29.

17.

17. Konishi H, Tanaka M, Takemura Y, Matsuzaki H, Ono Y, Kikkawa U, et al. Activation of protein kinase C by tyrosine phosphorylation in response to H2O2. Proc Natl Acad Sci USA 1997;94:11233-7.

18.

18. Ha H, Yu MR, Choi YJ, Lee HB. Activation of protein kinase c-delta and c-epsilon by oxidative stress in early diabetic rat kidney. Am J Kidney Dis 2001;38(4 Suppl 1):S204-7.

19.

19. Jung YS, Ryu BR, Lee BK, Mook-Jung I, Kim SU, Lee SH, et al. Role for PKC-epsilon in neuronal death induced by oxidative stress. Biochem Biophys Res Commun 2004;320:789-94.

20.

20. Kim HP, Wang X, Chen ZH, Lee SJ, Huang MH, Wang Y, et al. Autophagic proteins regulate cigarette smokeinduced apoptosis: protective role of heme oxygenase- 1. Autophagy 2008;4:887-95.

Tuberculosis & Respiratory Diseases

e-Submission OA Policy