Molecular and Cellular PharmacologyEffect of Staurosporine on Transcription Factor NF-κB in Human Keratinocytes
Section snippets
Materials
Staurosporine was obtained from the Sigma Chemical Co. The selective PKC inhibitor Ro32-0432 was synthesized by the Department of Medicinal Chemistry, SmithKline Beecham Pharmaceuticals, according to the reported procedure [17]. IL-1β and TNFα were prepared at SmithKline Beecham Pharmaceuticals as described previously 18, 19.
Cell Culture
Human foreskin keratinocytes were purchased from the Clonetics Corp. and grown as monolayers at 37°, 5% CO2 in KGM (Clonetics Corp.). KGM is based on the MCDB 153
Stimulation of IL-8 Production in Human Keratinocytes by Staurosporine Treatment
Studies aimed at investigating the mechanism(s) whereby IL-8 synthesis may be regulated in human keratinocytes revealed that production of this cytokine in response to stimulation with IL-1β was potentiated in response to treatment of the cells with the PKC inhibitor staurosporine [6]. Therefore, this phenomenon was investigated further by treating human keratinocytes with staurosporine in the absence of other inducers, such as IL-1β. Cultured human keratinocytes responded to treatment with
Discussion
The microbial alkaloid staurosporine is widely used as an inhibitor of PKC where it is proven to be a potent inhibitor of PMA-stimulated events, although staurosporine is known to be a fairly non-selective inhibitor [2]. Several recent studies report on the ability of staurosporine to potentiate a variety of cellular responses induced by the cytokines IL-1 and TNF 3, 4, 5, 6. Such observations have led investigators to speculate on the role of PKC in the signal transduction pathways induced by
References (39)
- et al.
Staurosporine, a potent inhibitor of phospholipid Ca2+ dependent protein kinase
Biochem Biophys Res Commun
(1986) - et al.
Protein kinase CIs its pivotal role in cellular activation over-stated?
Trends Pharmacol Sci
(1994) - et al.
Interleukin-8 production is regulated by protein kinase C in human keratinocytes
J Invest Dermatol
(1994) - et al.
Staurosporine-induced neurite outgrowth in PC12h cells
Exp Cell Res
(1989) - et al.
Studies on the regulatory mechanisms of interleukin-8 gene expression in resting and IFN-γ-treated neutrophilsEvidence on the capability of staurosporine of inducing the production of interleukin-8 by human neutrophils
Biochem Biophys Res Commun
(1993) - et al.
Protein kinases negatively affect nuclear factor-κB activation by tumor necrosis factor-α at two different stages in promyelocytic HL-60 cells
J Biol Chem
(1992) - et al.
Purification and characterization of human recombinant interleukin-1β
J Biol Chem
(1987) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Anal Biochem
(1976)- et al.
Staurosporine induces dissolution of microfilament bundles by a protein kinase C-independent pathway
Exp Cell Res
(1990) - et al.
Activation of DNA-binding activity in an apparently cytoplasmic precursor of the NF-κB transcription factor
Cell
(1988)
Identification and characterization of an IκB kinase
Cell
The IκB kinase complex (IKK) contains two kinase subunits, IKKα and IKKβ, necessary for IκB phosphorylation and NF-κB activation
Cell
Tyrosine phosphorylation of IκB-α activates NF-κB without proteolytic degradation of IκB-α
Cell
Regulation of HIV-1 long terminal repeats by interaction of C/EBP (NF-IL6) and NF-κB/Rel transcription factors
J Biol Chem
Contrasting effects of the protein kinase C inhibitor, staurosporine, on cytokine and phorbol ester stimulation of fructose 2,6-bisphosphate and prostaglandin E production by fibroblasts in vitro
Biochem J
Contrasting effects of the protein kinase C inhibitor staurosporine in the interleukin-1 and phorbol ester activation pathways in the EL4–6.1 thymoma cell line
J Cell Physiol
Staurosporine, but not Ro 31-8220, induces interleukin 2 production and synergizes with interleukin 1α in EL4 thymoma cells. Activation of nuclear factor κB as a common signal for staurosporine and interleukin 1α
Biochem J
Staurosporine, a non-specific PKC inhibitor, induces keratinocyte differentiation and raises intracellular calcium, but Ro31–8220 does not
J Cell Physiol
Staurosporine-induced differentiation in a neuroblastoma cell line, NB-1
Agric Biol Chem
Cited by (10)
Multiple kinases regulate mafA expression in the pancreatic beta cell line MIN6
2008, Archives of Biochemistry and BiophysicsCitation Excerpt :On the other hand, staurosporine also activates some transcription factors like NF-кB. As such, staurosporine increases the synthesis of some NF-кB target genes [37,38]. Therefore, perhaps staurosporine activates a transcription factor necessary for inducing mafA expression.
Contribution of protein kinase A and protein kinase C pathways in ultraviolet B-induced IL-8 expression by human keratinocytes
2005, CytokineCitation Excerpt :It has been shown that staurosporine can induce the translocation of PKCɛ isoforms [36] which in turn are involved in NF-κB activation [37]. In cultured human keratinocytes, NF-κB activation following staurosporine treatment is associated with an increase of IL-8 production [38]. These data are confirmed by our own results that indicated that staurosporine treatment of non-irradiated NCTC 2544 cells induced IL-8 activation.
Arsenic Induces Human Keratinocyte Apoptosis by the FAS/FAS Ligand Pathway, Which Correlates with Alterations in Nuclear Factor-κB and Activator Protein-1 Activity
2004, Journal of Investigative DermatologyCitation Excerpt :NF-κB regulates multiple and complex gene expression associated with both cell proliferation and death. Several keratinocyte-proliferation-associated factors are regulated by NF-κB, such as interleukin-8 (Mukaida et al, 1990;Tuschil et al, 1992;Chabot-Fletcher and Breton, 1998). NF-κB was reported to provide survival signals that prevent cells from apoptosis by Fas and TNF-receptor-initiated apoptotic signals (Beg and Baltimore, 1996;Liu et al, 1996).
Induction of cyclooxygenase-2 by staurosporine through the activation of nuclear factor for IL-6 (NF-IL6) and activator protein 2 (AP2) in an osteoblast-like cell line
2002, Biochemical PharmacologyCitation Excerpt :The mechanism responsible for these cellular events is not known, but staurosporine-mediated cellular processes are believed to be independent of its inhibitory activity. Staurosporine was shown to induce the expression of nitric oxide synthase and interleukins by activating NF-κB as described in many reports [7–10]. C/EBP family members may also be involved in staurosporine-mediated processes, as postulated by Hecker et al. [7].
Stimulation of catecholamine biosynthesis via the protein kinase C pathway by endothelin-1 in PC12 rat pheochromocytoma cells
2002, Biochemical PharmacologyCitation Excerpt :In the present study, we also showed that ET-1 significantly increased PKC activity in PC12 cells (Fig. 4). The increase in PKC activity stimulated by ET-1 was inhibited completely by Ro-32-0432, confirming that Ro-32-0432 is indeed a PKC inhibitor in our experimental system [25]. It should also be noted that the increases in TH enzyme activity and mRNA levels evoked by ET-1 stimulation were inhibited by 100 nM Ro-32-0432 (Figs. 1B and 2).