RRC ID 61492
Author Tominaga K, Higuchi K, Sasaki E, Suto R, Watanabe T, Fujiwara Y, Oshitani N, Matsumoto T, Kim S, Iwao H, Arakawa T.
Title Correlation of MAP kinases with COX-2 induction differs between MKN45 and HT29 cells.
Journal Aliment Pharmacol Ther
Abstract BACKGROUND:Mitogen-activated protein (MAP) kinases, including extracellular signal-regulated kinases (ERK),c-Jun NH2-terminal kinases (JNK) and p38 MAP kinase (p38 MAPK) are important intermediates of the signal-transduction pathway from the cell surface to the nucleus. Expression of cyclooxygenase (COX)-2, associated with proliferation, apoptosis or both of gastrointestinal cancer cells, is mediated through MAP kinase families. However, the correlation between respective MAP kinase signals and COX-2 in the proliferation of gastric and colon cancer cells has not been well elucidated.
AIM:We examined the effect of selective inhibitors of MAP kinases and COX-2 on serum-induced proliferation of gastric (MKN45) and colon (HT29) cancer cells.
METHODS:After 24-h serum starvation, cancer cells were stimulated with 2% serum and COX-2 inhibitors (NS398 10 micromol/L, or etodolac 100 micromol/L) or 1 h after preincubation with inhibitors for ERK (PD98059 20 micromol/L) or p38 MAPK (SB203580 10 micromol/L). Phosphorylated MAP kinases and COX-2 protein were evaluated by Western blotting, and the proliferation of cancer cells was estimated by 3H-thymidine incorporation. Transcription factors nuclear factor-kappaB and CREB were assayed by an electorophoretic mobility shift assay.
RESULTS:Serum increased the proliferation of MKN45 and HT29 cells by 280% and 200%, respectively, compared with the control levels (100%). In both cancer cells, phosphorylated MAP kinases were increased within 30 min after stimulation. PD98059 and SB203580 inhibited the serum-induced proliferation of MKN45 by 21% and 51% and of HT29 by 81% and 69%, respectively. NS398 and etodolac inhibited the proliferation of HT29 by 21% and 41%, respectively, but not that of MKN45. PD98059 and SB203580 also suppressed serum-induced expression of COX-2 protein in HT29 cells. In addition to the activation of MAP kinases and COX-2, activities of nuclear factor-kappaB and CREB were also increased during HT29 cell proliferation.
CONCLUSIONS:These results suggest that the correlation of MAP kinases with COX-2 induction for cell proliferation differs between MKN45 and HT29 cells.
Volume 20 Suppl 1
Pages 143-50
Published 2004-7-1
DOI 10.1111/j.1365-2036.2004.01986.x
PMID 15298620
MeSH Blotting, Western Cell Division Cell Line, Tumor Colonic Neoplasms / enzymology* Cyclooxygenase 2 Cyclooxygenase 2 Inhibitors Cyclooxygenase Inhibitors / pharmacology Enzyme Inhibitors / pharmacology Flavonoids / pharmacology HT29 Cells Humans Imidazoles / pharmacology Isoenzymes / antagonists & inhibitors Isoenzymes / metabolism* Membrane Proteins Mitogen-Activated Protein Kinase Kinases / metabolism* Prostaglandin-Endoperoxide Synthases / metabolism* Pyridines / pharmacology Serum / physiology Stomach Neoplasms / enzymology*
IF 7.515
Human and Animal Cells MKN45(RCB1001)