A role for focal adhesion kinase signaling in tumor necrosis factor-alpha-dependent matrix metalloproteinase-9 production in a cholangiocarcinoma cell line, CCKS1.
We have previously reported that tumor necrosis factor-alpha (TNF-alpha) stimulation of CCKS1, a cell line established from cholangiocarcinoma with i.p. dissemination, dramatically increased matrix metalloproteinase-9 (MMP-9) production and tumor invasion. We investigated the role of focal adhesion kinase (FAK) in TNF-alpha-dependent production of MMP-9 in CCKS1 and FAK-null mouse fibroblast cells. TNF-alpha stimulation of CCKS1 or wild-type fibroblasts substantially activated FAK phosphorylation and increased MMP-9 production. In contrast, FAK-null fibroblasts could not respond well to TNF-alpha stimulation. Conditional expression of wild-type FAK in FAK-null cells restored the TNF-alpha-dependent production of MMP-9. TNF-alpha treatment activated the kinase activity of FAK and its phosphorylation especially at Y397 and Y925. Phosphorylated FAK accumulated at focal adhesions and formed a complex with growth factor receptor binding protein 2 and SOS. In contrast, Y397F FAK and Y925F FAK, whose Y397 and Y925 were replaced with phenylalanine, respectively, as well as KD FAK, whose kinase was inactivated, could not restore the MMP-9 production. In addition, small interfering RNA against FAK drastically suppressed the TNF-alpha-dependent production of MMP-9 and inhibited the TNF-alpha-dependent invasion of CCKS1. Taken together, our results suggest the pivotal role of FAK in TNF-alpha-dependent production of MMP-9 and subsequent activation of tumor invasion.
Mon
Naing Naing
NN
Division of Cancer Biology, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, 466 Nagoya, Japan.
Hasegawa
Hitoki
H
Thant
Aye Aye
AA
Huang
Pengyu
P
Tanimura
Yoko
Y
Senga
Takeshi
T
Hamaguchi
Michinari
M
D000818:Animals; D041702:BALB 3T3 Cells; D001650:Bile Duct Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002448:Cell Adhesion / Q000502:physiology; D045744:Cell Line, Tumor; D018281:Cholangiocarcinoma / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D051380:GRB2 Adaptor Protein / Q000378:metabolism; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis*; D051379:Mice; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D020839:Son of Sevenless Protein, Drosophila / Q000378:metabolism; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism* / Q000494:pharmacology
D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D034741:RNA, Small Interfering; D020839:Son of Sevenless Protein, Drosophila; D014409:Tumor Necrosis Factor-alpha; D051416:Focal Adhesion Protein-Tyrosine Kinases; D020780:Matrix Metalloproteinase 9