ATP sensitizes H460 lung carcinoma cells to cisplatin-induced apoptosis.
Platinum resistance of cancer cells may evolve due to a decrease in intracellular drug accumulation, decreased cell permeability or by an increased deactivation of the drug by glutathione (GSH). The aim of this study was (1) to investigate the effect of adenosine 5'-triphosphate (ATP) on the cytotoxicity of cisplatin in a large cell lung carcinoma cell line (H460), and (2) to examine the potential involvement of increased cisplatin uptake, GSH depletion and pyrimidine starvation by ATP in this effect. H460 cells were harvested and seeded (5% CO(2); 37 degrees C). Subsequently, cells were incubated with medium or ATP followed by an incubation with cisplatin. Cytotoxicity screening was analyzed by the sulforhodamine B (SRB) colorimetric assay, lactate dehydrogenase and caspase-3/7 activity. Pre-incubation for 72h with 0.3 and 3mM ATP strongly enhanced the anti-proliferative potency of cisplatin 2.9- and 7.6-fold, respectively. Moreover, after incubation of H460 cells with 0.3mM ATP the intracellular platinum concentration increased, indicating increased cisplatin uptake by ATP. ATP, despite lowering the LD(50) of cisplatin, did not modulate GSH levels in H460 cells. ATP itself showed a biphasic effect on H460 cell growth: 0.3mM inhibited H460 cell growth via the pyrimidine starvation effect, activation of caspase-3/7 and LDH leakage, while 3mM ATP showed no effect on cell growth. In conclusion, ATP sensitizes the H460 cells to cisplatin-induced apoptosis. The effect of 0.3mM ATP is not due to GSH depletion but involves increased cisplatin uptake and pyrimidine starvation due to ATP conversion to adenosine followed by cellular uptake.
Department of Epidemiology, NUTRIM School for Nutrition, Toxicology and Metabolism, Maastricht University Medical Centre Maastricht, The Netherlands. e.swennen@farmaco.unimaas.nl <e.swennen@farmaco.unimaas.nl>
Els L R
EL
Swennen
Vanessa
V
Ummels
Irina
I
Buss
Ulrich
U
Jaehde
Aalt
A
Bast
Pieter C
PC
Dagnelie
D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000633:toxicity*; D017209:Apoptosis; D002277:Carcinoma / Q000188:drug therapy / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000633:toxicity*; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D045022:Lactate Dehydrogenases / Q000378:metabolism; D008175:Lung Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D011743:Pyrimidines / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism
D000970:Antineoplastic Agents; D011743:Pyrimidines; D012235:Rhodamines; C022027:lissamine rhodamine B; D000255:Adenosine Triphosphate; D045022:Lactate Dehydrogenases; D005982:Glutathione Transferase; D053148:Caspase 3; D053179:Caspase 7; C030986:pyrimidine; D002945:Cisplatin