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Nuclear Factor-kappaB Regulates Cyclooxygenase-2 Expression and Cell Proliferation in Human Gastric Cancer Cells

 Joo Weon Lim  ;  Hyeyoung Kim  ;  Kyung Hwan Kim 
 LABORATORY INVESTIGATION, Vol.81(3) : 349-360, 2001 
Journal Title
Issue Date
6-Ketoprostaglandin F1 alpha/metabolism ; Adenocarcinoma* ; Cell Division/drug effects ; Cell Division/physiology ; Cyclooxygenase 2 ; Cyclooxygenase 2 Inhibitors ; Cyclooxygenase Inhibitors/pharmacology ; DNA-Binding Proteins/analysis ; DNA-Binding Proteins/genetics ; Dinoprostone/pharmacology ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; I-kappa B Proteins* ; Isoenzymes/genetics* ; Isoenzymes/metabolism ; Membrane Proteins ; Mutagenesis ; NF-KappaB Inhibitor alpha ; NF-kappa B/metabolism* ; Nitrobenzenes/pharmacology ; Oligonucleotides, Antisense/pharmacology ; Prostaglandin-Endoperoxide Synthases/genetics* ; Prostaglandin-Endoperoxide Synthases/metabolism ; RNA, Messenger/analysis ; Stomach Neoplasms* ; Sulfonamides/pharmacology ; Thromboxane B2/metabolism ; Transfection ; Tumor Cells, Cultured/cytology ; Tumor Cells, Cultured/enzymology
Nuclear factor-kappaB (NF-kappaB) is a transcriptional regulator of inducible expression of genes including cyclooxygenase-2 (COX-2), regulating cell proliferation. NF-kappaB is kept silent in the cytoplasm via interaction with the inhibitory protein IkappaBalpha and transmigrated into the nucleus upon activation. However, constitutive NF-kappaB has been found in the nucleus of some cancer cells. We investigated the role of NF-kappaB in COX-2 expression and cell proliferation in human gastric cancer AGS cells. AGS cells were treated with antisense oligodeoxynucleotide (AS ODN) or sense oligodeoxynucleotide (S ODN) for the NF-kappaB subunit p50, or they were transfected with a mutated IkappaBalpha gene (MAD-3 mutant) or a control vector, pcDNA-3. AGS cells were treated with COX-2 inhibitors such as indomethacine and NS-398 or prostaglandin E2. mRNA expression for COX-2, and protein levels for p50, IkappaBalpha, and COX-2 were determined by reverse transcription polymerase chain reaction and Western blot analysis. The NF-kappaB levels were examined by electrophoretic mobility shift assay. Thromboxane B2 (TXB2) and 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha) levels were determined by enzyme-linked immunosorbent assay. Cell proliferation was assessed by viable cell counting, [3H] thymidine incorporation, and colony formation. The nuclear level of p50 decreased in AGS cells treated with AS ODN. The IkappaBa mutant was observed in cells transfected with the mutated IkappaBa gene. NF-kappaB was inhibited in cells treated with AS ODN or transfected with the mutated IkappaBalpha gene, compared with the cells treated with S ODN or transfected with control vector. Cell proliferation, mRNA expression and protein level of COX-2, and production of TXB2 and 6-keto-PGF1alpha were inhibited in cells treated with AS ODN or transfected with the mutated IkappaBalpha gene, which had lower NF-kappaB levels than cells treated with S ODN or transfected with control vector. COX-2 inhibitors suppressed cell proliferation and production of TXB2 and 6-keto-PGF1alpha, in a dose-dependant manner. Prostaglandin E2 prevented the inhibition of proliferation in cells treated with AS ODN or transfected with the mutated IkappaBalpha gene. In conclusion, NF-kappaB mediates COX-2 expression, which may be related to cell proliferation, in human gastric cancer cells.
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1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
5. Research Institutes (연구소) > Institute of Gastroenterology (소화기병연구소) > 1. Journal Papers
Yonsei Authors
Kim, Kyung Hwan(김경환)
Lim, Joo Weon(임주원)
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