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Long-term incubation with proteasome inhibitors (PIs) induces IκBα degradation via the lysosomal pathway in an IκB kinase (IKK)-dependent and IKK-independent manner.

Identifieur interne : 000214 ( Ncbi/Checkpoint ); précédent : 000213; suivant : 000215

Long-term incubation with proteasome inhibitors (PIs) induces IκBα degradation via the lysosomal pathway in an IκB kinase (IKK)-dependent and IKK-independent manner.

Auteurs : Kyoung-Hee Lee ; Jiyeong Jeong ; Chul-Gyu Yoo

Source :

RBID : pubmed:24085292

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English descriptors

Abstract

Proteasome inhibitors (PIs) have been reported to induce apoptosis in many types of tumor. Their apoptotic activities have been suggested to be associated with the up-regulation of molecules implicated in pro-apoptotic cascades such as p53, p21(Waf1), and p27(Kip1). Moreover, the blocking of NF-κB nuclear translocation via the stabilization of IκB is an important mechanism of PI-induced apoptosis. However, we found that long-term incubation with PIs (PS-341 or MG132) increased NF-κB-regulated gene expression such as COX-2, cIAP2, XIAP, and IL-8 in a dose- and time-dependent manner, which was mediated by phosphorylation of IκBα and its subsequent degradation via the alternative route, lysosome. Overexpression of the IκBα superrepressor (IκBα-SR) blocked PI-induced NF-κB activation. Treatment with lysosomal inhibitors (ammonium chloride or chloroquine) or inhibitors of cathepsins (Z-FF-FMK or Z-FA-FMK) or knock-down of LC3B expression by siRNAs suppressed PI-induced IκBα degradation. Furthermore, we found that both IKK-dependent and IKK-independent pathways were required for PI-induced IκBα degradation. Pretreatment with IKKβ specific inhibitor, SC-514, partially suppressed IκBα degradation and IL-8 production by PIs. Blockade of IKK activity using insolubilization by heat shock (HS) and knock-down by siRNAs for IKKβ only delayed IκBα degradation up to 8 h after treatment with PIs. In addition, PIs induced Akt-dependent inactivation of GSK-3β. Inactive GSK-3β accelerated PI-induced IκBα degradation. Overexpression of active GSK-3β (S9A) or knock-down of GSK-3β delayed PI-induced IκBα degradation. Collectively, our data demonstrate that long-term incubation with PIs activates NF-κB, which is mediated by IκBα degradation via the lysosome in an IKK-dependent and IKK-independent manner.

DOI: 10.1074/jbc.M113.480921
PubMed: 24085292


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Le document en format XML

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<term>Dipeptides (pharmacology)</term>
<term>Gene Expression Regulation (drug effects)</term>
<term>Gene Expression Regulation (genetics)</term>
<term>Glycogen Synthase Kinase 3 (genetics)</term>
<term>Glycogen Synthase Kinase 3 (metabolism)</term>
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<term>I-kappa B Proteins (genetics)</term>
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<term>Interleukin-8 (biosynthesis)</term>
<term>Interleukin-8 (genetics)</term>
<term>Ketones (pharmacology)</term>
<term>Leupeptins (pharmacology)</term>
<term>Lysosomes (genetics)</term>
<term>Lysosomes (metabolism)</term>
<term>Microtubule-Associated Proteins (genetics)</term>
<term>Microtubule-Associated Proteins (metabolism)</term>
<term>Mutation, Missense</term>
<term>NF-KappaB Inhibitor alpha</term>
<term>NF-kappa B (metabolism)</term>
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<term>Proteolysis (drug effects)</term>
<term>Pyrazines (pharmacology)</term>
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<term>Acides boroniques (pharmacologie)</term>
<term>Antinéoplasiques (pharmacologie)</term>
<term>Bortézomib</term>
<term>Cétones (pharmacologie)</term>
<term>Dipeptides (pharmacologie)</term>
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<term>Facteurs temps</term>
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<term>Glycogen Synthase Kinase 3 (métabolisme)</term>
<term>Glycogen synthase kinase 3 beta</term>
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<term>I-kappa B Kinase (génétique)</term>
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<term>Protéines associées aux microtubules (métabolisme)</term>
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<term>Pyrazines (pharmacologie)</term>
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<term>Régulation de l'expression des gènes (génétique)</term>
<term>Thiophènes (pharmacologie)</term>
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<term>I-kappa B Kinase</term>
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<term>Microtubule-Associated Proteins</term>
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<term>Dipeptides</term>
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<term>Humans</term>
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<term>Facteurs temps</term>
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<term>Humains</term>
<term>Inhibiteur alpha de NF-KappaB</term>
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<div type="abstract" xml:lang="en">Proteasome inhibitors (PIs) have been reported to induce apoptosis in many types of tumor. Their apoptotic activities have been suggested to be associated with the up-regulation of molecules implicated in pro-apoptotic cascades such as p53, p21(Waf1), and p27(Kip1). Moreover, the blocking of NF-κB nuclear translocation via the stabilization of IκB is an important mechanism of PI-induced apoptosis. However, we found that long-term incubation with PIs (PS-341 or MG132) increased NF-κB-regulated gene expression such as COX-2, cIAP2, XIAP, and IL-8 in a dose- and time-dependent manner, which was mediated by phosphorylation of IκBα and its subsequent degradation via the alternative route, lysosome. Overexpression of the IκBα superrepressor (IκBα-SR) blocked PI-induced NF-κB activation. Treatment with lysosomal inhibitors (ammonium chloride or chloroquine) or inhibitors of cathepsins (Z-FF-FMK or Z-FA-FMK) or knock-down of LC3B expression by siRNAs suppressed PI-induced IκBα degradation. Furthermore, we found that both IKK-dependent and IKK-independent pathways were required for PI-induced IκBα degradation. Pretreatment with IKKβ specific inhibitor, SC-514, partially suppressed IκBα degradation and IL-8 production by PIs. Blockade of IKK activity using insolubilization by heat shock (HS) and knock-down by siRNAs for IKKβ only delayed IκBα degradation up to 8 h after treatment with PIs. In addition, PIs induced Akt-dependent inactivation of GSK-3β. Inactive GSK-3β accelerated PI-induced IκBα degradation. Overexpression of active GSK-3β (S9A) or knock-down of GSK-3β delayed PI-induced IκBα degradation. Collectively, our data demonstrate that long-term incubation with PIs activates NF-κB, which is mediated by IκBα degradation via the lysosome in an IKK-dependent and IKK-independent manner. </div>
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