Extracellular ATP induces apoptotic signaling in human monocyte leukemic cells, HL-60 and F-36P.
Extracellular adenosine 5'-triphosphate (ATP) affects the function of many tissues and cells. To confirm the biological activity of ATP on human myeloid leukemic cells, F-36P and HL-60, cells were treated with a variety of concentrations of ATP. The stimulation with extracellular ATP induced the arrest of cell proliferation and cell death from the analysis of Annexin-V staining and caspase activity by flow cytometry. The Annexin-V positive cells in both cell lines were dramatically increased following ATP stimulation. The expression of P2 purinergic receptor genes was confirmed, such as P2X1, P2X4, P2X5, P2X7 and P2Y1, P2Y2, P2Y4, P2Y5, P2Y6, P2Y11 in both leukemic cell lines. Interestingly, ATP induced intracellular calcium flux in HL-60 cells but not in F-36P cells, as determined by Fluo-3 AM staining. Cell cycle analysis revealed that ATP treatment arrested both F-36P and HL-60 cells at G1/GO. Taken together, these data showed that extracellular ATP via P2 receptor genes was involved in the cell proliferation and survival in human myeloid leukemic cells, HL-60 and F-36P cells by the induction of apoptosis and control of cell cycle. Our data suggest that treatment with extracellular nucleotides may be a novel and powerful therapeutic avenue for myeloid leukemic disease.
Yoon
Mi-Jung
MJ
Cell and Gene Therapy Research Institute, Graduate School of Life Science and Biotechnology, Pochon CHA University CHA General Hospital, Seoul 135-081, Korea.
Lee
Hae-Jin
HJ
Kim
Jae-Hwan
JH
Kim
Dong-Ku
DK
D000255:Adenosine Triphosphate / Q000008:administration & dosage / Q000494:pharmacology*; D017302:Annexins / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003124:Colorimetry; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D007951:Leukemia, Myeloid / Q000473:pathology*; D018048:Receptors, Purinergic P2 / Q000187:drug effects; D058473:Receptors, Purinergic P2Y2; D015398:Signal Transduction / Q000187:drug effects*; D013778:Tetrazolium Salts; D013844:Thiazoles
D017302:Annexins; C546385:P2RY2 protein, human; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; D013778:Tetrazolium Salts; D013844:Thiazoles; D000255:Adenosine Triphosphate; D020169:Caspases; C022616:thiazolyl blue; D002118:Calcium