Acrolein causes inhibitor κb-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells

Noel D. Horton, Shyam Biswal, Lucindra L. Corrigan, Julie Bratta, James P. Kehrer

Research output: Contribution to journalArticle

Abstract

Acrolein is a highly electrophilic α,β-unsaturated aldehyde to which humans are exposed in various situations. In the present study, the effects of sublethal doses of acrolein on nuclear factor κB (NF-κB) activation in A549 human lung adenocarcinoma cells were investigated. Immediately following a 30-min exposure to 45 fmol of acrolein/cell, glutathione (GSH) and DNA synthesis and NF-κB binding were reduced by more than 80%. All parameters returned to normal or supranormal levels by 8 h post-treatment. Pretreatment with acrolein completely blocked 12-O-tetradecanoylphorbol-13-acetate (TPA)- induced activation of NF-κB. Cells treated for 1 h with 1 mM diethyl maleate (DEM) showed a 34 and 53% decrease in GSH and DNA synthesis, respectively. DEM also reduced NF-κB activation by 64% at 2 h posttreatment, with recovery to within 22% of control at 8 h. Both acrolein and DEM decreased NF-κB function ~50% at 2 h after treatment with TPA, as shown by a secreted alkaline phosphatase reporter assay. GSH returned to control levels by 8 h after DEM treatment, but proliferation remained significantly depressed for 24 h. Interestingly, DEM caused a profound decrease in NF-κB binding, even at doses as low as 0.125 mM that had little effect on GSH. Neither acrolein nor DEM had any effect on the levels of phosphorylated or nonphosphorylated inhibitor κB-α (IκB-α). Furthermore, acrolein decreased NF-κB activation in cells depleted of IκB-α by TPA stimulation in the presence of cycloheximide, demonstrating that the decrease in NF-κB activation was not the result of increased binding by the inhibitory protein. This conclusion was further supported by the finding that acrolein modified NF-κB in the cytosol prior to chemical dissociation from IκB with detergent. Together, these data support the conclusion that the inhibition of NF-κB activation by acrolein and DEM is IκB-independent. The mechanism appears to be related to direct modification of thiol groups in the NF-κB subunits.

Original languageEnglish (US)
Pages (from-to)9200-9206
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number14
DOIs
StatePublished - Apr 2 1999
Externally publishedYes

Fingerprint

diethyl maleate
Acrolein
Chemical activation
Tetradecanoylphorbol Acetate
Acetates
A549 Cells
Adenocarcinoma of lung
Fibroblast Growth Factors
DNA
Level control
Cycloheximide
Sulfhydryl Compounds
Aldehydes
Detergents
Cytosol
Glutathione
Alkaline Phosphatase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Acrolein causes inhibitor κb-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells. / Horton, Noel D.; Biswal, Shyam; Corrigan, Lucindra L.; Bratta, Julie; Kehrer, James P.

In: Journal of Biological Chemistry, Vol. 274, No. 14, 02.04.1999, p. 9200-9206.

Research output: Contribution to journalArticle

Horton, Noel D. ; Biswal, Shyam ; Corrigan, Lucindra L. ; Bratta, Julie ; Kehrer, James P. / Acrolein causes inhibitor κb-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 14. pp. 9200-9206.
@article{b601da4ac2424a6a9369c89404fb8406,
title = "Acrolein causes inhibitor κb-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells",
abstract = "Acrolein is a highly electrophilic α,β-unsaturated aldehyde to which humans are exposed in various situations. In the present study, the effects of sublethal doses of acrolein on nuclear factor κB (NF-κB) activation in A549 human lung adenocarcinoma cells were investigated. Immediately following a 30-min exposure to 45 fmol of acrolein/cell, glutathione (GSH) and DNA synthesis and NF-κB binding were reduced by more than 80{\%}. All parameters returned to normal or supranormal levels by 8 h post-treatment. Pretreatment with acrolein completely blocked 12-O-tetradecanoylphorbol-13-acetate (TPA)- induced activation of NF-κB. Cells treated for 1 h with 1 mM diethyl maleate (DEM) showed a 34 and 53{\%} decrease in GSH and DNA synthesis, respectively. DEM also reduced NF-κB activation by 64{\%} at 2 h posttreatment, with recovery to within 22{\%} of control at 8 h. Both acrolein and DEM decreased NF-κB function ~50{\%} at 2 h after treatment with TPA, as shown by a secreted alkaline phosphatase reporter assay. GSH returned to control levels by 8 h after DEM treatment, but proliferation remained significantly depressed for 24 h. Interestingly, DEM caused a profound decrease in NF-κB binding, even at doses as low as 0.125 mM that had little effect on GSH. Neither acrolein nor DEM had any effect on the levels of phosphorylated or nonphosphorylated inhibitor κB-α (IκB-α). Furthermore, acrolein decreased NF-κB activation in cells depleted of IκB-α by TPA stimulation in the presence of cycloheximide, demonstrating that the decrease in NF-κB activation was not the result of increased binding by the inhibitory protein. This conclusion was further supported by the finding that acrolein modified NF-κB in the cytosol prior to chemical dissociation from IκB with detergent. Together, these data support the conclusion that the inhibition of NF-κB activation by acrolein and DEM is IκB-independent. The mechanism appears to be related to direct modification of thiol groups in the NF-κB subunits.",
author = "Horton, {Noel D.} and Shyam Biswal and Corrigan, {Lucindra L.} and Julie Bratta and Kehrer, {James P.}",
year = "1999",
month = "4",
day = "2",
doi = "10.1074/jbc.274.14.9200",
language = "English (US)",
volume = "274",
pages = "9200--9206",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "14",

}

TY - JOUR

T1 - Acrolein causes inhibitor κb-independent decreases in nuclear factor κB activation in human lung adenocarcinoma (A549) cells

AU - Horton, Noel D.

AU - Biswal, Shyam

AU - Corrigan, Lucindra L.

AU - Bratta, Julie

AU - Kehrer, James P.

PY - 1999/4/2

Y1 - 1999/4/2

N2 - Acrolein is a highly electrophilic α,β-unsaturated aldehyde to which humans are exposed in various situations. In the present study, the effects of sublethal doses of acrolein on nuclear factor κB (NF-κB) activation in A549 human lung adenocarcinoma cells were investigated. Immediately following a 30-min exposure to 45 fmol of acrolein/cell, glutathione (GSH) and DNA synthesis and NF-κB binding were reduced by more than 80%. All parameters returned to normal or supranormal levels by 8 h post-treatment. Pretreatment with acrolein completely blocked 12-O-tetradecanoylphorbol-13-acetate (TPA)- induced activation of NF-κB. Cells treated for 1 h with 1 mM diethyl maleate (DEM) showed a 34 and 53% decrease in GSH and DNA synthesis, respectively. DEM also reduced NF-κB activation by 64% at 2 h posttreatment, with recovery to within 22% of control at 8 h. Both acrolein and DEM decreased NF-κB function ~50% at 2 h after treatment with TPA, as shown by a secreted alkaline phosphatase reporter assay. GSH returned to control levels by 8 h after DEM treatment, but proliferation remained significantly depressed for 24 h. Interestingly, DEM caused a profound decrease in NF-κB binding, even at doses as low as 0.125 mM that had little effect on GSH. Neither acrolein nor DEM had any effect on the levels of phosphorylated or nonphosphorylated inhibitor κB-α (IκB-α). Furthermore, acrolein decreased NF-κB activation in cells depleted of IκB-α by TPA stimulation in the presence of cycloheximide, demonstrating that the decrease in NF-κB activation was not the result of increased binding by the inhibitory protein. This conclusion was further supported by the finding that acrolein modified NF-κB in the cytosol prior to chemical dissociation from IκB with detergent. Together, these data support the conclusion that the inhibition of NF-κB activation by acrolein and DEM is IκB-independent. The mechanism appears to be related to direct modification of thiol groups in the NF-κB subunits.

AB - Acrolein is a highly electrophilic α,β-unsaturated aldehyde to which humans are exposed in various situations. In the present study, the effects of sublethal doses of acrolein on nuclear factor κB (NF-κB) activation in A549 human lung adenocarcinoma cells were investigated. Immediately following a 30-min exposure to 45 fmol of acrolein/cell, glutathione (GSH) and DNA synthesis and NF-κB binding were reduced by more than 80%. All parameters returned to normal or supranormal levels by 8 h post-treatment. Pretreatment with acrolein completely blocked 12-O-tetradecanoylphorbol-13-acetate (TPA)- induced activation of NF-κB. Cells treated for 1 h with 1 mM diethyl maleate (DEM) showed a 34 and 53% decrease in GSH and DNA synthesis, respectively. DEM also reduced NF-κB activation by 64% at 2 h posttreatment, with recovery to within 22% of control at 8 h. Both acrolein and DEM decreased NF-κB function ~50% at 2 h after treatment with TPA, as shown by a secreted alkaline phosphatase reporter assay. GSH returned to control levels by 8 h after DEM treatment, but proliferation remained significantly depressed for 24 h. Interestingly, DEM caused a profound decrease in NF-κB binding, even at doses as low as 0.125 mM that had little effect on GSH. Neither acrolein nor DEM had any effect on the levels of phosphorylated or nonphosphorylated inhibitor κB-α (IκB-α). Furthermore, acrolein decreased NF-κB activation in cells depleted of IκB-α by TPA stimulation in the presence of cycloheximide, demonstrating that the decrease in NF-κB activation was not the result of increased binding by the inhibitory protein. This conclusion was further supported by the finding that acrolein modified NF-κB in the cytosol prior to chemical dissociation from IκB with detergent. Together, these data support the conclusion that the inhibition of NF-κB activation by acrolein and DEM is IκB-independent. The mechanism appears to be related to direct modification of thiol groups in the NF-κB subunits.

UR - http://www.scopus.com/inward/record.url?scp=0033515441&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033515441&partnerID=8YFLogxK

U2 - 10.1074/jbc.274.14.9200

DO - 10.1074/jbc.274.14.9200

M3 - Article

C2 - 10092592

AN - SCOPUS:0033515441

VL - 274

SP - 9200

EP - 9206

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 14

ER -