Protein Kinase Cα but not p44/42 Mitogen-Activated Protein Kinase, p38, or c-Jun NH2-Terminal Kinase Is Required for Intercellular Adhesion Molecule-1 Expression Mediated by Interleukin-1β: Involvement of Sequential Activation of Tyrosine Kinase, Nuclear Factor-κB-Inducing Kinase, and IκB Kinase 2
Abstract
IL-1β induced an increase in ICAM-1 expression in human A549 epithelial cells and immunofluorescence staining confirmed this result. Tyrosine kinase inhibitors (genistein or tyrphostin 23) or phosphatidylcholine-specific phospholipase C inhibitor (D609) attenuated IL-1β-induced ICAM-1 expression. IL-1β produced an increase in PKC activity and this effect was abolished by D609. PKC inhibitors (staurosporine, Ro 31-8220, calphostin C, or Go 6976) also inhibited IL-1β-induced response. TPA, a PKC activator, stimulated ICAM-1 expression as well, this effect being inhibited by tyrosine kinase inhibitors. Treatment of cells with IL-1β resulted in stimulation of p44/42 MAPK, p38, and JNK. However, neither the mitogen activated protein kinase kinase inhibitor PD 98059 nor the p38 inhibitor SB 203580 affected IL-1β-induced ICAM-1 expression. NF-κB DNA-protein binding and ICAM-1 promoter activity were enhanced by IL-1β and these effects were inhibited by tyrphostin 23, but not by PD 98059 or SB 203580. TPA also stimulated NF-κB DNA-protein binding and ICAM-1 promoter activity as well, these effects being inhibited by tyrosine kinase inhibitors. Dominant-negative PKCα, NIK, or IKK2, but not IKK1 mutant, inhibited IL-1β- or TPA-induced ICAM-1 promoter activity. IKK activity was stimulated by either IL-1β or TPA, and these effects were inhibited by Ro 31-8220 or tyrphostin 23. Taken together, IL-1β activates phosphatidylcholine-specific phospholipase C and induces activation of PKCα and protein tyrosine kinase, resulting in the stimulation of NIK, IKK2, and NF-κB in the ICAM-1 promoter, then initiation of ICAM-1 expression. However, activation of p44/42 MAPK, p38, and JNK is not involved.
Footnotes
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Send reprint requests to: Dr. Ching-Chow Chen, Institute of Pharmacology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, 1st Section, Taipei 10018, Taiwan. E-mail:ccchen{at}ha.mc.ntu.edu.tw
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This work was supported by a research grant from the National Science Council of Taiwan.
- Abbreviations:
- ICAM-1
- intercellular adhesion molecule-1
- IL-1
- interleukin-1
- IL-1RI
- interleukin-1 type I receptor
- TNF
- tumor necrosis factor
- NF-κB
- nuclear factor-κB
- NIK
- nuclear factor-κB-inducing kinase
- IKK
- IκB kinase
- MAPK
- mitogen-activated protein kinase
- JNK
- c-Jun NH2-terminal kinase
- PKC
- protein kinase C
- PC-PLC
- phosphatidylcholine-specific phospholipase C
- ERK
- extracellular signal-regulated kinase
- DMEM
- Dulbecco's modified Eagle's medium
- FCS
- fetal calf serum
- TPA
- 12-O-tetradecanoylphorbol-13-acetate
- PDTC
- pyrrolidine dithiocarbamate
- PAGE
- polyacrylamide gel electrophoresis
- ECL
- enhanced chemiluminescence
- ELISA
- enzyme-linked immunosorbent assay
- TTBS
- Tween-20/Tris-buffered saline
- PMSF
- phenylmethylsulfonyl fluoride
- EMSA
- electrophoretic mobility shift assay
- DTT
- dithiothreitol
- MEK
- mitogen-activated protein kinase kinase
- DAG
- diacylglycerol
- LPS
- lipopolysaccharide
- GST
- glutathioneS-transferase
- MBP
- myelin basic protein
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- Received March 24, 2000.
- Accepted September 8, 2000.
- The American Society for Pharmacology and Experimental Therapeutics
