Induction of DNA damage, inhibition of DNA synthesis and suppression of c-myc expression by the anthracycline analog, idarubicin (4-demethoxy-daunorubicin) in the MCF-7 breast tumor cell line

Purpose: Studies were designed to elucidate the basis for the antiproliferative activity of the anthracycline Antibiotic, idarubicin (4-demethoxy-daunorubicin) in MCF-7 breast tumor cells.

Methods: Growth inhibition was evaluated using the MTT tetrazolium dye assay, induction of DNA strand breaks was determined by alkaline elution, inhibition of DNA synthesis was assessed by measuring the incorporation of labelled thymidine into DNA, modulation of the expression of the c-Myc oncogene was determined by Northern blotting and the induction of Apoptosis was evaluated by alkaline unwinding, static field gel electrophoresis, terminal end labelling and assessment of cell morphology.

Results: MCF-7 cells were relatively sensitive to idarubicin, with an IC50 value for growth inhibition of approximately 0.01 microM. While DNA strand breakage was not evident below a concentration of 0.1 microM idarubicin, where growth inhibition exceeded 70%, both the inhibition of DNA synthesis and suppression of c-Myc expression closely paralleled the profile of antiproliferative activity for idarubicin. Finally, while exposure to idarubicin resulted in a substantial loss of viable cells within 48-72 h, there was no morphological evidence of apoptotic body formation. The absence of Apoptosis in cells exposed to idarubicin was supported by studies demonstrating the absence of DNA fragmentation using gel electrophoresis, alkaline elution and in situ DNA end-labelling assays.

Conclusions: The results of these studies extend previous results from this laboratory indicating an association between suppression of c-Myc expression, inhibition of DNA synthesis and growth arrest by Topoisomerase II inhibitors, as well as the lack of induction of apoptotic cell death by Topoisomerase II inhibitors in MCF-7 breast tumor cells.