p53-Independent induction of Fas and apoptosis in leukemic cells by an adenosine derivative, Cl-IB-MECA.
A(3) adenosine receptor (A(3)AR) agonists have been reported to influence cell death and survival. The effects of an A(3)AR agonist, 1-[2-chloro-6-[[(3-iodophenyl)methyl]amino]-9H-purin-9-yl]-1-deoxy-N-methyl-beta-D-ribofuranonamide (Cl-IB-MECA), on apoptosis in two human leukemia cell lines, HL-60 and MOLT-4, were investigated. Cl-IB-MECA (> or =30 microM) increased the apoptotic fractions, as determined using fluorescence-activated cell sorting (FACS) analysis, and activated caspase 3 and poly-ADP-ribose-polymerase. Known messengers coupled to A(3)AR (phospholipase C and intracellular calcium) did not seem to play a role in the induction of apoptosis. Neither dantrolene nor BAPTA-AM affected the Cl-IB-MECA-induced apoptosis. Cl-IB-MECA failed to activate phospholipase C in HL-60 cells, while UTP activated it through endogenous P2Y(2) receptors. Induction of apoptosis during a 48hr exposure to Cl-IB-MECA was not prevented by the A(3)AR antagonists [5-propyl-2-ethyl-4-propyl-3-(ethylsulfanylcarbonyl)-6-phenylpyridine-5-carboxylate] (MRS 1220) or N-[9-chloro-2-(2-furanyl)[1,2,4]triazolo[1,5-c]quinazolin-5-yl]benzeneacetamide (MRS 1523). Furthermore, higher concentrations of MRS 1220, which would also antagonize A(1) and A(2A) receptors, were ineffective in preventing the apoptosis. Although Cl-IB-MECA has been shown in other systems to cause apoptosis through an A(3)AR-mediated mechanism, in these cells it appeared to be an adenosine receptor-independent effect, which required prolonged incubation. In both HL-60 and MOLT-4 cells, Cl-IB-MECA induced the expression of Fas, a death receptor. This induction of Fas was not dependent upon p53, because p53 is not expressed in an active form in either HL-60 or MOLT-4 cells. Cl-IB-MECA-induced apoptosis in HL-60 cells was augmented by an agonistic Fas antibody, CH-11, and this increase was suppressed by the antagonistic anti-Fas antibody ZB-4. Therefore, Cl-IB-MECA induced apoptosis via a novel, p53-independent up-regulation of Fas.
Kim
Seong Gon
SG
Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Ravi
Gnana
G
Hoffmann
Carsten
C
Jung
Yun Jin
YJ
Kim
Min
M
Chen
Aishe
A
Jacobson
Kenneth A
KA
D000241:Adenosine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000906:Antibodies / Q000494:pharmacology; D017209:Apoptosis; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D004347:Drug Interactions; D018922:HL-60 Cells; D006801:Humans; D007938:Leukemia / Q000473:pathology; D058915:Purinergic P1 Receptor Antagonists; D043684:Receptor, Adenosine A3; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*; D019014:fas Receptor / Q000096:biosynthesis* / Q000276:immunology
D000906:Antibodies; D058915:Purinergic P1 Receptor Antagonists; D043684:Receptor, Adenosine A3; D016159:Tumor Suppressor Protein p53; D019014:fas Receptor; D010738:Type C Phospholipases; D000241:Adenosine; D002118:Calcium; C090034:2-chloro-N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide