Apoptosis, induced in tumors by anticancer agents, is characterized by caspase activation, impaired cellular respiration and decreased cellular ATP. Respiration in Jurkat and HL-60 cells treated with doxorubicin, dactinomycin or platinum drugs is measured using a Pd(II) phosphor that monitors [O2] in cell suspensions as a function of time. Cellular ATP is determined using the luciferin-luciferase bioluminescence system. Intracellular caspase activation is measured by allowing caspases to cleave Ac-DEVD-AFC to the fluorescent AFC, which is detected on HPLC. Comparing the ways in which respiration, ATP level, and caspase activity vary with time points up differences between the mechanisms of actions of doxorubicin, dactinomycin and the platinum compounds. These methods accurately determine the sensitivity of tumors to anticancer drugs.
Keywords: Apoptosis, cellular ATP, 7-amino-4-trifluoromethyl coumarin, pan-caspase inhibitor, doxorubicin
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