Multiparametric investigations for studying the time-dependence of the effect of Adriamycin on some human leukemic lymphoblasts using multiwavelength videomicrofluorometry

摘要

Rationale investigations of the cytotoxic processes of drugs on heterogeneous cell populations point out the need to get information on individual cells. Such information, including both dose-effects and time-effects, can be obtained through a multiparametric approach involving multiwavelength microfluorometry and numerical image analysis. For that purpose, cells can be simultaneously stained with Hoechst 33342 (nuclear staining), Rhodamine 123 (mitochondrial staining) and Nile Red (cell contour delineation). As a first step, that protocol was used to look for differences between a sensitive lymphoblastoid cell line (CEM-WT) and two related resistant ones (CEM-VLB and CEM-VM1) which differ both by their resistance level and by the nature of the mechanism involved in the drug-resistance. Adriamycin was used to check both dose and time effects. Changes in the biological parameters accessible from R123 and Ho33342 labeling were found more informative than changes in morphometric parameters. They allow the monitoring of the time dependent cellular modifications induced by the cytotoxic process. Moreover the different mechanisms involved in the cellular resistance can be differentiated. That protocol will be used in the near future to study the resistance induced by newly available cytotoxic drugs extracted from marine animals.