Glial cell line-derived neurotrophic factor enhances nuclear factor-kappaB activity and invasive potential in human pancreatic cancer cells.
OBJECTIVE
The invasive potential is increased by glial cell line-derived neurotrophic factor (GDNF) in human pancreatic cancer cell lines. We researched whether the signaling pathway activated by GDNF correlates with the nuclear factor-kappaB (NF-kappaB) in human pancreatic cancer cell lines and whether the inhibition of NF-kappaB activity is associated with suppression of invasive potential.


METHODS
Proliferation of human pancreatic cancer cell lines (BxPC-3 and MIA PaCa-2) was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays (MTT assay). NF-kappaB activity was examined by dual luciferase assay and electrophoretic mobility shift assay. In addition, to investigate the invasive potential, an in vitro invasion assay was performed.


RESULTS
Proliferation of both cell lines was decreased by a proteasome inhibitor, MG132, in a dose-dependent manner, but proliferation of control and IkappaBalphaM vector-transfected BxPC-3 cells was similar. The invasion cell number and the NF-kappaB activity were increased by GDNF stimulation. However, in the presence of MG132 or IkappaBalphaM, which blocks the nuclear localization of NF-kappaB, both were significantly suppressed. Furthermore, reduced activity of both remained unchanged by GDNF stimulation.


CONCLUSIONS
These results indicate that GDNF promotes NF-kappaB activation and that the latter is involved in the invasive potential of human pancreatic cancer cells.
Takahashi
Hiroki
H
Department of Gastroenterological Surgery, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. takahasi@med.nagoya-cu.ac.jp
Funahashi
Hitoshi
H
Sawai
Hirozumi
H
Sakamoto
Masaki
M
Matsuo
Yoichi
Y
Yamamoto
Minoru
M
Okada
Yuji
Y
Hayakawa
Tetsushi
T
Manabe
Tadao
T
D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D003094:Collagen; D004338:Drug Combinations; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D017930:Genes, Reporter; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics / Q000502:physiology; D007797:Laminin; D007976:Leupeptins / Q000494:pharmacology; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000502:physiology*; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D009414:Nerve Growth Factors / Q000494:pharmacology*; D010190:Pancreatic Neoplasms / Q000473:pathology*; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins / Q000502:physiology; D011994:Recombinant Proteins / Q000494:pharmacology; D015398:Signal Transduction; D015533:Transcriptional Activation / Q000187:drug effects*; D014162:Transfection
D004338:Drug Combinations; C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D020672:I-kappa B Proteins; D007797:Laminin; D007976:Leupeptins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D009363:Neoplasm Proteins; D009414:Nerve Growth Factors; D011509:Proteoglycans; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C059485:matrigel; D000072000:NF-KappaB Inhibitor alpha; D003094:Collagen; D008156:Luciferases; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde