Effect of cigarette smoke extract on the proliferation of human airway epithelial cells and expression and activation of FAK

Abstract

The effect of cigarette smoke extract (CSE) on the proliferation of human airway epithelial cells and the possible mechanism was studied. After airway epithelial cells were treated with different concentrations of CSE for 24 h, the cell proliferation was measured by MTT and the distribution of different cell cycles by flow cytometry. The FAK expression level was detected by Western blot and the degree of tyrosine phosphorylation by immunoprecipitation. The results showed that CSE could inhibit the proliferation of human airway epithelial cells, arrest the epithelial cells in G1 phase of cell cycle, dramatically decrease the number of epithelial cells in S and G2 phases; Mean-while CSE could decrease the expression level of FAK and the degree of its tyrosine phosphorylation. The above effects of CSE were concentration-dependent. The expression of FAK and the degree of its phosphorylation was positively correlated to the increased number of epithelial cells in G1 phase, and negatively to the number of epithelial cells in S and G2 phases. It was concluded that the mechanism by which CSE could inhibit the proliferation of human epithelial cells was contributed to the increased expression and activation of FAK.

Keywords

cigarette smoke extract / airway epithelial cell / proliferations / FAK

References

[1] AnneL H, DenisP, JocelyneH, et al.. Fibronectin and its α_sβ₁-integrin receptor are involved in the wound-repair process of airway epithelium. Am J Physiol, 1996, 271(5 Pt 1): L726-L726

[2] ZahmJ M, KaplanH, HerardA L, et al.. Cell migration and proliferation during the in vitro wound repair of the respiratory epithelium. Cell Motil Cytoskeleton, 1997, 37(1): 33-33

[3] NakamuraY, RombergerD J, TateL, et al.. Cigarette smoke inhibits lung fibroblast proliferation and chemotaxis. Am J Respir Crit Care Med, 1995, 151: 1497-1497

[4] HellermannG R, NagyS B, LongX Y, et al.. Mechanism of cigarette smoke condensate-induced acute inflammatory response in human bronchial epithelial cells. Respir Res J, 2002, 3(1): 22-22

[5] HamannK J, DorscheidD R, ConfortiA E, et al.. Expression of Fas (CD95) and FasL (CD95L) in human airway epithelium. Am J Respir Cell Mol Biol, 1998, 19(4): 537-537

[6] EduardoP S, EnriqueR. Src family kinases are required for integrin-mediated but not for G Protein-coupled receptor stimulation of focal adhesion kinase autophosphorylation at Tyr-397. J Biol Chem, 2001, 276(2): 17788-17788

[7] DucskoL, CarolineH D, TadashiY, et al.. Focal adhesion kinase: at the crossroads of signal transduction. J cell science, 1997, 110: 401-401

[8] van de WaterBob, NagelkerkeJ F, StevensJ L, et al.. Dephosphorylation of Focal Adhesion Kinase (FAK) and Loss of Focal Contacts Precede Caspase-mediated Cleavage of FAK during Apoptosis in Renal Epithelial Cells. J Biol Chem, 1999, 274(9): 13328-13328

[9] HeatherA W, JohnM W, PamelaJ G, et al.. Perlecan Up-Regulation of FRNK Suppresses Smooth Muscle Cell Proliferation via Inhibition of FAK Signaling. Mol Biol Cell, 2003, 14(5): 1941-1941