Extracellular ATP induces apoptotic signaling in human monocyte leukemic cells, HL-60 and F-36P

Abstract

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.

Aqueous Cinnamon Extract (ACE-c) from the bark of Cinnamomum cassiacauses apoptosis in human cervical cancer cell line (SiHa) through loss of mitochondrial membrane potential

“…Intracellular Ca 2+ levels were analyzed in control cells as well as cells treated with different doses of ACE- c : (0-80 μg/ml) by flow cytometry [29]. Cells were loaded with 5 μM Fluo-3/AM (Sigma, St. Louis, MO) and 100 μg/ml of Pluronic F127 (Sigma, St. Louis, MO) in centrifuge tubes and incubated at 37°C, 5% CO 2 for 1 h in the dark.…”

Purinergic signalling and cancer

“…P2Y 2 and P2Y 11 receptors were upregulated during granulocytic differentiation of HL-60 cells [455]. ATP induced apoptotic cell death of both HL-60 and F-36P leukaemia cell lines [456]. Static magnetic field exposure of HL-60 cells increased the increase in [Ca 2+ ] i produced by ATP [457,458].…”

Hematopoietic modulators as potential agents for the treatment of leukemia

“…High concentrations of extracellular ATP (> 1 mM) have shown cytotoxic activity in the K562 cell line and its multidrug resistant counterpart, Lucena-1, by a still non-identified receptor (66). In addition, ATP also induces apoptosis in leukemic HL60 and F-36P cells (67).…”