Thrombin induces NF-kappaB activation and IL-8/CXCL8 expression in lung epithelial cells by a Rac1-dependent PI3K/Akt pathway.
We previously showed that thrombin induces interleukin (IL)-8/CXCL8 expression via the protein kinase C (PKC)α/c-Src-dependent IκB kinase α/β (IKKα/β)/NF-κB signaling pathway in human lung epithelial cells. In this study, we further investigated the roles of Rac1, phosphoinositide 3-kinase (PI3K), and Akt in thrombin-induced NF-κB activation and IL-8/CXCL8 expression. Thrombin-induced IL-8/CXCL8 release and IL-8/CXCL8-luciferase activity were attenuated by a PI3K inhibitor (LY294002), an Akt inhibitor (1-L-6-hydroxymethyl-chiro-inositol-2-((R)-2-O-methyl-3-O-octadecylcarbonate)), and the dominant negative mutants of Rac1 (RacN17) and Akt (AktDN). Treatment of cells with thrombin caused activation of Rac and Akt. The thrombin-induced increase in Akt activation was inhibited by RacN17 and LY294002. Stimulation of cells with thrombin resulted in increases in IKKα/β activation and κB-luciferase activity; these effects were inhibited by RacN17, LY294002, an Akt inhibitor, and AktDN. Treatment of cells with thrombin induced Gβγ, p85α, and Rac1 complex formation in a time-dependent manner. These results imply that thrombin activates the Rac1/PI3K/Akt pathway through formation of the Gβγ, Rac1, and p85α complex to induce IKKα/β activation, NF-κB transactivation, and IL-8/CXCL8 expression in human lung epithelial cells.
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 110, Taiwan.
Chien-Huang
CH
Lin
Hui-Wen
HW
Cheng
Hon-Ping
HP
Ma
Chih-Hsiung
CH
Wu
Chuang-Ye
CY
Hong
Bing-Chang
BC
Chen
D019943:Amino Acid Substitution; D000818:Animals; D002417:Cattle; D002460:Cell Line; D002867:Chromones / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics / Q000378:metabolism; D007294:Inositol / Q000031:analogs & derivatives / Q000494:pharmacology; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000235:genetics; D008168:Lung; D009025:Morpholines / Q000494:pharmacology; D020125:Mutation, Missense; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013917:Thrombin / Q000378:metabolism* / Q000494:pharmacology; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
C477931:1L-6-hydroxymethyl-chiro-inositol 2(R)-2-O-methyl-3-O-octadecylcarbonate; C508606:CXCL8 protein, human; D002867:Chromones; D004791:Enzyme Inhibitors; D016209:Interleukin-8; D009025:Morpholines; D016328:NF-kappa B; C107953:RAC1 protein, human; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D007294:Inositol; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D051550:I-kappa B Kinase; D013917:Thrombin; D020830:rac1 GTP-Binding Protein
Circ Res. 2002 Sep 6;91(5):398-405
12215488
J Cell Sci. 2006 Apr 15;119(Pt 8):1528-36
16551696
J Mol Cell Cardiol. 2004 Aug;37(2):449-71
15276015
Carcinogenesis. 2010 Apr;31(4):587-96
20106902
J Biol Chem. 1998 May 22;273(21):12779-85
9582304
J Biol Chem. 2006 Feb 10;281(6):3210-6
16339142
Immunity. 2000 Jan;12(1):7-16
10661401
Respiration. 1996;63(1):42-8
8833992
Am J Respir Cell Mol Biol. 2004 Nov;31(5):517-27
15284075
Pharmacol Ther. 2007 Jul;115(1):70-83
17532472
Nature. 2000 Sep 14;407(6801):258-64
11001069
J Biol Chem. 2001 Aug 10;276(32):30188-98
11402028
Am J Physiol. 1993 Apr;264(4 Pt 1):L413-8
8476069
J Biol Chem. 2005 Nov 4;280(44):36952-61
16120607
Exp Mol Med. 2008 Apr 30;40(2):167-75
18446055
Genes Dev. 1997 Sep 15;11(18):2295-322
9308960
Mol Pharmacol. 2004 Sep;66(3):561-71
15322248
Curr Biol. 1999 Jun 3;9(11):601-4
10359702
J Biol Chem. 2003 Oct 10;278(41):39413-21
12888567
Thromb Haemost. 2008 Dec;100(6):1021-8
19132225
J Thromb Haemost. 2007 Dec;5(12):2484-93
17883592
Curr Opin Immunol. 2000 Feb;12(1):52-8
10679399
J Biol Chem. 2003 Jan 3;278(1):400-6
12393875
Biochem J. 1996 May 1;315 ( Pt 3):775-9
8645157
Mol Immunol. 2008 May;45(9):2446-54
18258304
J Immunol. 1989 Aug 15;143(4):1366-71
2663993
Am J Physiol Cell Physiol. 2007 Jan;292(1):C362-71
16914532
Exp Lung Res. 1991 Jan-Feb;17(1):17-23
2013270
J Immunol. 2006 Sep 1;177(5):3427-38
16920985
J Immunol. 2002 Apr 1;168(7):3577-85
11907122
Science. 2002 May 31;296(5573):1655-7
12040186
J Biol Chem. 2000 Sep 29;275(39):29955-9
10893407
Mol Cell Biol. 2001 Aug;21(16):5554-65
11463837
Mol Cell Biol. 1996 Dec;16(12):7115-21
8943367
Biochem Biophys Res Commun. 2008 Feb 22;366(4):1030-5
18086557
J Immunol. 2006 Jul 1;177(1):681-93
16785567
J Cell Biochem. 2009 Oct 15;108(3):716-25
19693771
J Leukoc Biol. 2002 Nov;72(5):847-55
12429706
Br J Pharmacol. 2004 Sep;143(2):269-75
15302675
J Immunol. 2003 May 1;170(9):4767-75
12707358
J Leukoc Biol. 2007 Sep;82(3):619-29
17586662
Nature. 1999 Sep 2;401(6748):86-90
10485711
Science. 1997 Jan 31;275(5300):665-8
9005852
Trends Cell Biol. 2005 Mar;15(3):163-71
15752980
J Biol Chem. 1994 May 6;269(18):13289-95
8175759
EMBO J. 1996 Dec 2;15(23):6541-51
8978681
J Immunol. 2009 Oct 1;183(7):4682-92
19734226
Genes Dev. 1997 Feb 15;11(4):463-75
9042860
Curr Drug Targets Inflamm Allergy. 2005 Dec;4(6):619-25
17305518
Mol Cell Biol. 1999 Sep;19(9):6333-44
10454580
J Thromb Haemost. 2005 Oct;3(10):2296-306
16194206
Cell. 1997 Feb 21;88(4):435-7
9038334
Biochem Biophys Res Commun. 2010 Jun 4;396(3):748-54
20451499
Endocr Rev. 2005 Feb;26(1):1-43
15689571
Mol Cell Biol. 1999 Mar;19(3):2180-8
10022904
Blood. 2000 Feb 1;95(3):959-64
10648409