Mitochondria do not play a major role in landomycin E-induced ROS burst and circumvention of multiple drug resistance in HL-60 leukemia cells.

Aim. To study the molecular mechanisms of reactive oxygen species (ROS) involvement in circumventing the cancer drug resistance by novel angucycline antibiotic landomycin E in HL-60 human leukemia cells and its drug-resistant sublines HL-60/adr and HL-60/vinc. Methods. MTT assay, trypan blue exclusion test, DCFDA and JC-1 staining of cells. Results. Landomycin E (LE) leads to a massive hydrogen peroxide production in HL-60 cell line already 1h after the drug addition to the cell culture, while depolarization of mitochondria is observed only at 6-12h, which indicates on the extra-mitochondrial ROS production by LE. The drug-resistant cells of HL-60/vinc (P-gp+) despite 100-fold resistance to doxorubicin (Dx) action, demonstrated no difference in the resistance to LE compared to the parental cell line, while HL-60/adr line (MRP-1+), which was found to be 200-fold resistant to Dx action, had shown a weak (2-fold) decrease in sensitivity to LE. Circumvention of drug resistance by LE in HL-60/adr cells was accompanied by a 2-fold higher level of H2O2 compared to the wild-type cells, but the mitochondrial respiratory chain inhibitors had no impact on this phenomenon. Conclusions. LE-induced cell death is accompanied by massive hydrogen peroxide production, consisting of two peaks - major one at 1h and secondary at 12h after drug treatment. The highest ROS production was observed in HL-60/adr cells, which have shown a 2-fold increase of resistance to LE compared to parental cells. However, mitochondria seem to play secondary role in this process, due to the fact that use of specific inhibitors of mitochondrial respiratory chain did not affect the early ROS burst, induced by LE in tumor cells. [ABSTRACT FROM AUTHOR]