The use of bioluminescence to evaluate the influence of chemotherapeutic drugs on ATP-levels of malignant cell lines.

Abstract

Influence of various cytostatic agents on intracellular ATP concentrations of malignant cell lines was studied. The HL-60 promyelocytic cell line, the SW-480 cell line, derived from an adenocarcinoma of the colon and the SW-620 cell line, derived from a lymphonodal metastasis of the colon carcinoma, were investigated. Cell lines were incubated with cytostatic agents and changes in intracellular ATP concentrations were measured after various incubation times by means of bioluminescence. A marked fall in intracellular ATP concentrations was observed, when HL-60 cells were incubated with drugs used in clinical protocols for treatment of acute leukaemia (daunorubicin, vinblastine, vincristine), whereas only a slight decrease of ATP concentrations was measured after incubation with bleomycin, dacarbazin and prednisolone. The decrease in intracellular ATP concentrations of SW-480 and SW-620 cells was much less pronounced after incubation with cytostatic agents compared to the HL-60 cells. This is in accordance with the clinical experience of the known resistance of colon carcinoma against cytostatic agents. Dose-response curves were obtained for the single cytostatic agents. Comparison of intracellular ATP concentrations and cell viability as determined by the trypan blue and eosin dye exclusion test showed that the trypan blue dye exclusion test underestimated cell kill compared to the eosin dye exclusion test and the bioluminescence assay.