論文 - 詳細
| RRC ID | 18926 |
|---|---|
| 著者 | Yaguchi T, Saito M, Yasuda Y, Kanno T, Nakano T, Nishizaki T. |
| タイトル | Higher concentrations of extracellular ATP suppress proliferation of Caco-2 human colonic cancer cells via an unknown receptor involving PKC inhibition. |
| ジャーナル | Cell Physiol Biochem |
| Abstract |
BACKGROUND/AIMS:Adenosine 5'-triphosphate (ATP) mediates a variety of signal transductions via ATP receptors such as P2X and P2Y receptors. The present study aimed at understanding the mechanism underlying extracellular ATP-induced suppression of Caco-2 human colonic cancer cell proliferation. METHODS:Caco-2 cells were cultured. To examine cell viability and cell cycling, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, fluorescent cytochemistry, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and flow cytometry were carried out. To see mRNA expression of ATP receptors, reverse transcription-polymerase chain reaction (RT-PCR) was performed. To examine PKC activity and mitogen-activated protein (MAP) kinase activity, in situ PKC assay and Western blotting using an anti-extracellular signal-regulated kinase 1 (ERK1)-antibody and an anti-phospho-ERK antibody were carried out. RESULTS:Extracellular ATP or the unhydrolyzed ATP analogue 5'-adenylyimido-diphosphate (AMP-PNP) reduced Caco-2 cell viability in a concentration (10 microM-10 mM)-dependent manner at 48-h treatment, and the effect was not affected by caspase inhibitors. Caco-2 cells were little reactive to propidium iodide and Hoechst 33342 or little positive to TUNEL after 48-h treatment with ATP (1 mM). In the flow cytometry, 48-h treatment with ATP (1 mM) arrested cell cycling at the S phase in Caco-2 cells. P(2) purinoceptor agonists reduced Caco-2 cell viability with the order of potency: 2-methylthio ATP>UTP>beta, gamma-methylene ATP, and the ATP effect was partially inhibited by suramin, a non-selective inhibitor of P(2) purinoceptors. The PKC inhibitor GF109203X or the MAP kinase kinase inhibitor PD98059 reduced Caco-2 cell viability to an extent similar to that achieved by ATP (1 mM), and no further reduction was obtained with co-treatment with ATP. ATP and its ATP analogues such as AMP-PNP and ATPgammaS, at higher concentrations (1-10 mM), inhibited PKC activation in Caco-2 cells in a fashion that mimics the effect of GF109203X, but PD98059 exhibited no effect on PKC activation. The inhibitory effect of ATP on PKC activation was not found with SK-N-SH cells, a human neuroblastoma cell line, but the cells expressed all the mRNAs for P2X and P2Y receptors that Caco-2 cells did. ATP (10 mM) or GF109203X inhibited activation of ERK, a MAP kinase, in Caco-2 cells. CONCLUSION:Extracellular ATP, at higher concentrations, suppresses Caco-2 cell proliferation at the S phase of cell cycling by inhibiting PKC, possibly as mediated via an unknown ATP receptor, followed by MAP kinase. |
| 巻・号 | 26(2) |
| ページ | 125-34 |
| 公開日 | 2010-1-1 |
| DOI | 10.1159/000320518 |
| PII | 000320518 |
| PMID | 20798496 |
| MeSH | Adenosine Triphosphate / analogs & derivatives Adenosine Triphosphate / pharmacology* Antineoplastic Agents / pharmacology* Caco-2 Cells Cell Proliferation / drug effects Cell Survival Colonic Neoplasms / enzymology* Colonic Neoplasms / metabolism Colonic Neoplasms / pathology Flavonoids / pharmacology Humans Indoles / pharmacology Maleimides / pharmacology Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors Mitogen-Activated Protein Kinase 3 / metabolism Protein Kinase C / antagonists & inhibitors* Protein Kinase C / metabolism Purinergic P2 Receptor Agonists / pharmacology Receptors, Purinergic P2X / chemistry Receptors, Purinergic P2X / metabolism Receptors, Purinergic P2Y / chemistry Receptors, Purinergic P2Y / metabolism |
| IF | 5.5 |
| 引用数 | 17 |
| WOS 分野 | PHYSIOLOGY CELL BIOLOGY |
| リソース情報 | |
| ヒト・動物細胞 | CACO-2(RCB0988) SK-N-SH(RCB0426) |