Phosphorylation of extracellular signal-regulated kinase 1/2, p38 mitogen-activated protein kinase and nuclear translocation of nuclear factor-kappaB are involved in upregulation of matrix metalloproteinase-9 by tumour necrosis factor-alpha.
BACKGROUND
Upregulation of matrix metalloproteinase-9 (MMP-9) induced by tumour necrosis factor-alpha (TNF-alpha) is reportedly involved in a variety of non-neoplastic and neoplastic diseases. In this study, we examined which signalling pathways are involved in TNF-alpha-induced MMP-9 upregulation in cholangiocarcinoma (CC).


METHODS
We used two CC cell lines: HuCCT-1 and CCKS-1.


RESULTS
In an ex vivo study using HuCCT-1 and CCKS-1 cells, TNF-alpha treatment induced MMP-9 production and activation via interaction with TNF receptor-1 (TNF-R1) but not with TNF receptor-2 (TNF-R2), shown by zymography, and increased MMP-9 promoter activity (luciferase assay). As for the signalling pathway, TNF-alpha stimulation led to the phosphorylation of extracellular signal-regulated kinase 1/2 (Erk1/2) and p38 mitogen-activated protein kinase (p38MAPK) and translocation of nuclear factor kappaB (NF-kappaB) (p65) into the nuclei. Inhibition studies using SB203580 (inhibitor of p38MAPK), U0126 (inhibitor of mitogen-activated or extracellular signal-regulated protein kinase 1/2) and MG132 (inhibitor of NF-kappaB) showed that the phosphorylation of Erk1/2 and p38MAPK with activation of NF-kappaB was closely related to MMP-9 upregulation in both cell lines.


CONCLUSIONS
These data suggest that TNF-alpha/TNF-R1 interaction leads to the phosphorylation of Erk1/2 and p38MAPK and nuclear translocation of NF-kappaB, which is closely associated with the production and activation of MMP-9 in cultured CC cells of HuCTT-1 and CCKS-1. Upregulation of MMP-9 with NF-kappaB activation may be involved in the tumour invasion of CC.
Itatsu
Keita
K
Department of Human Pathology, Kanazawa University Graduate School of Medicine, Kanazawa, Japan.
Sasaki
Motoko
M
Harada
Kenichi
K
Yamaguchi
Junpei
J
Ikeda
Hiroko
H
Sato
Yasunori
Y
Ohta
Tetsuo
T
Sato
Hiroshi
H
Nagino
Masato
M
Nimura
Yuji
Y
Nakanuma
Yasuni
Y
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D018281:Cholangiocarcinoma / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008156:Luciferases; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D016328:NF-kappa B / Q000502:physiology; D010766:Phosphorylation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D015854:Up-Regulation; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D017931:DNA Primers; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D008156:Luciferases; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; D020780:Matrix Metalloproteinase 9