Acute cadmium exposure enhances AP-1 DNA binding and induces cytokines expression and heat shock protein 70 in HepG2 cells.
Cadmium (Cd) has been regarded as one of the inflammation-related xenobiotics. Cd has been extensively studied in many cellular systems, but a lot of parameters have been evaluated in different experimental conditions. This study was undertaken to examine the effects of low cadmium concentrations in HepG2 cells in the oxidative stress produced, the IL-1beta, tumor necrosis factor (TNF-alpha), IL-6, and IL-8 expression, production of heat shock protein 70 (Hsp70) and the activation of nuclear factors activation protein-1 (AP-1) and NF-kappaB under the same experimental conditions. Also, the participation of TNF-alpha and oxidative stress in AP-1 activation was evaluated. Lipid peroxidation damage increased 1.5 times after the first hour of Cd treatment and increased 1.9 times after 2h. Similar values were maintained until 6h. Reduced glutathione (GSH) diminished 65% after 6h CdCl(2) treatment. N-acetylcysteine (NAC) pre-treatment increased 332% GSH in Cd-treated cells. RNA was isolated from HepG2 cells after 0.5, 1, 3, or 6h incubation with 1, 5, or 10 microM CdCl(2). TNF-alpha and IL-1beta presented a maximum response after 1h treatment, while IL-6 and IL-8 maximum response was after 3h treatment. The Hsp70, determined by Western blot, was constitutively produced, and it increased after 3h Cd treatment. NF-kappaB activation, determined by EMSA, was not increased as a result of Cd treatment. DNA binding of AP-1 was detected and increased, with time up to 4h with an increment of 24 times control value with 5 microM CdCl(2). The HepG2 cells were pretreated with anti-TNF-alpha antibody or 1mM N-acetylcysteine 1h before Cd treatment. Anti-TNF-alpha treatment reduced 67% AP-1 activation, while NAC 47.5%. These data indicate that, Cd-induced TNF-alpha and IL-1beta, that probably, activate AP-1 transcription factor and IL-6 and IL-8 were induced. Anti-TNF-alpha and NAC partially inhibited AP-1 activation. All imply that, a number of factors participate in AP-1 cadmium-induced activation. The Hsp70 is produced by the HepG2 cells after cadmium treatment, and probably has a role in the non-participation of NF-kappaB in the cellular response.
Souza
Verónica
V
Departamento de Ciencias de la Salud, División de Ciencias Biológicas y de la Salud, Universidad Autónoma Metropolitana-Iztapalapa, Colonia La Vicentina, México, D.F. 09340, Mexico.
Escobar Md
Ma del Carmen
Mdel C
Gómez-Quiroz
Luis
L
Bucio
Leticia
L
Hernández
Elizabeth
E
Cossio
Edmundo Chávez
EC
Gutiérrez-Ruiz
Ma Concepción
MC
D019256:Cadmium Chloride / Q000633:toxicity*; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis*; D006801:Humans; D008054:Lipid Peroxides / Q000378:metabolism; D008113:Liver Neoplasms / Q000473:pathology; D011485:Protein Binding; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D016207:Cytokines; D018840:HSP70 Heat-Shock Proteins; D008054:Lipid Peroxides; D012333:RNA, Messenger; D018808:Transcription Factor AP-1; D004247:DNA; D019256:Cadmium Chloride