Epidermal growth factor promotes invasiveness of pancreatic cancer cells through NF-kappaB-mediated proteinase productions.
OBJECTIVE
Overexpression of epidermal growth factor (EGF) and EGF receptor has been associated with progression and invasive phenotype in pancreatic cancer. However, the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has been poorly understood. In this study, we determined the effect of EGF on the invasiveness and the related regulatory mechanism in two pancreatic cancer cell lines NOR-P1 and KP4.


METHODS
Invasion assay was performed to examine the invasiveness of tumor cells induced by EGF, and the expression of matrix metalloproteinase (MMP)-9, MMP-2, and plasminogen activator (uPA) was determined by reverse transcription-polymerase chain reaction and Western blot. Gelatin zymography was used to detect the activities of MMP-9 and MMP-2, and the nuclear factor-kappaB (NF-kappaB) binding activity was determined by electrophoretic mobility shift assay (EMSA).


RESULTS
EGF significantly increased the invasiveness of both cell lines but did not affect cell proliferation or adhesion. Increased invasiveness was associated with the induction of MMP-9 and uPA at both mRNA and protein levels. Furthermore, EGF stimulated NF-kappaB binding activity. Moreover, pretreatment of cells with NF-kappaB inhibitors, pyrrolidine dithiocarbamate or ibuprofen, markedly attenuated EGF-induced NF-kappaB activation. Subsequently, the EGF-induced MMP-9 and uPA expression and MMP-9 activity, as well as cell invasiveness, were also inhibited by these NF-kappaB inhibitors.


CONCLUSIONS
Our findings indicated that NF-kappaB-mediated MMP-9 and uPA induction was responsible for EGF-induced invasiveness in these pancreatic cancer cell lines and implicate that such anti-NF-kappaB therapy as the use of NF-kappaB inhibitors may contribute to the reduction of invasiveness of pancreatic cancer.
Zhang
Hao
H
Department of Surgery, The First Affiliated Hospital of China Medical University, Shenyang, China. haozhang@mail.cmu.edu.cn
Ma
Gang
G
Dong
Ming
M
Zhao
Meifen
M
Shen
Xiaodong
X
Ma
Zhigang
Z
Guo
Kejian
K
D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D006801:Humans; D020782:Matrix Metalloproteinases / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D010190:Pancreatic Neoplasms / Q000201:enzymology / Q000473:pathology*; D010447:Peptide Hydrolases / Q000235:genetics
D016328:NF-kappa B; D004815:Epidermal Growth Factor; D010447:Peptide Hydrolases; D020782:Matrix Metalloproteinases