PMID- 20798496
OWN - NLM
STAT- MEDLINE
DCOM- 20101208
LR  - 20190212
IS  - 1421-9778 (Electronic)
IS  - 1015-8987 (Linking)
VI  - 26
IP  - 2
DP  - 2010
TI  - Higher concentrations of extracellular ATP suppress proliferation of Caco-2 human 
      colonic cancer cells via an unknown receptor involving PKC inhibition.
PG  - 125-34
LID - 10.1159/000320518 [doi]
AB  - 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.
CI  - Copyright 2010 S. Karger AG, Basel.
FAU - Yaguchi, Takahiro
AU  - Yaguchi T
AD  - Division of Bioinformation, Department of Physiology, Hyogo College of Medicine, 
      Nishinomiya, Japan.
FAU - Saito, Masaru
AU  - Saito M
FAU - Yasuda, Yoshiyuki
AU  - Yasuda Y
FAU - Kanno, Takeshi
AU  - Kanno T
FAU - Nakano, Takashi
AU  - Nakano T
FAU - Nishizaki, Tomoyuki
AU  - Nishizaki T
LA  - eng
PT  - Journal Article
DEP - 20100824
PL  - Germany
TA  - Cell Physiol Biochem
JT  - Cellular physiology and biochemistry : international journal of experimental 
      cellular physiology, biochemistry, and pharmacology
JID - 9113221
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Flavonoids)
RN  - 0 (Indoles)
RN  - 0 (Maleimides)
RN  - 0 (Purinergic P2 Receptor Agonists)
RN  - 0 (Receptors, Purinergic P2X)
RN  - 0 (Receptors, Purinergic P2Y)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
RN  - EC 2.7.11.13 (Protein Kinase C)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 3)
RN  - L79H6N0V6C (bisindolylmaleimide I)
RN  - SJE1IO5E3I (2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one)
SB  - IM
MH  - Adenosine Triphosphate/analogs & derivatives/*pharmacology
MH  - Antineoplastic Agents/*pharmacology
MH  - Caco-2 Cells
MH  - Cell Proliferation/drug effects
MH  - Cell Survival
MH  - Colonic Neoplasms/*enzymology/metabolism/pathology
MH  - Flavonoids/pharmacology
MH  - Humans
MH  - Indoles/pharmacology
MH  - Maleimides/pharmacology
MH  - Mitogen-Activated Protein Kinase 3/antagonists & inhibitors/metabolism
MH  - Protein Kinase C/*antagonists & inhibitors/metabolism
MH  - Purinergic P2 Receptor Agonists/pharmacology
MH  - Receptors, Purinergic P2X/chemistry/metabolism
MH  - Receptors, Purinergic P2Y/chemistry/metabolism
EDAT- 2010/08/28 06:00
MHDA- 2010/12/14 06:00
CRDT- 2010/08/28 06:00
PHST- 2010/06/22 00:00 [accepted]
PHST- 2010/08/28 06:00 [entrez]
PHST- 2010/08/28 06:00 [pubmed]
PHST- 2010/12/14 06:00 [medline]
AID - 000320518 [pii]
AID - 10.1159/000320518 [doi]
PST - ppublish
SO  - Cell Physiol Biochem. 2010;26(2):125-34. doi: 10.1159/000320518. Epub 2010 Aug 
      24.