Aclarubicin-induced ROS generation and collapse of mitochondrial membrane potential in human cancer cell lines.
The cytotoxicity of aclarubicin (ACL) in A549 (human non-small lung), HepG2 (human hepatoma) and MCF-7 (human breast adenocarcinoma) cancer cell lines was evaluated and compared with that of doxorubicin (DOX). Changes in mitochondrial transmembrane potential (DeltaPsim), and production of reactive oxygen species (ROS) of drug-treated cells were monitored. Moreover, morphological changes associated with apoptosis were examined using double staining with Hoechst 33258-propidium iodide (PI). The results showed that ACL was much more cytotoxic than DOX in all investigated cell lines. Furthermore, ACL induced a concentration- and time-dependent increase in ROS production and decrease in mitochondrial membrane potential. The drugs, especially ACL, also induced ROS mediated apoptosis and necrosis pathways in all cell lines depending on the length of the post-treatment time. All these processes were partially inhibited by the antioxidants: N-acetylcysteine (NAC) and alpha-tocopherol. Of both drugs, DOX caused considerably weaker depolarization of the mitochondrial membrane. Its 10-fold higher concentration, as compared to ACL, was required to induce a similar effect, in accordance with the highly distinct cytotoxicity of these drugs towards investigated cells. In conclusion, ROS production preceded a decrease in mitochondrial membrane potential, but only changes in DeltaPsim were correlated with drug cytotoxicity in particular cell line. These results suggest that the impairment of DeltaPsim and an increase in ROS level might be important mechanisms of ACL cytotoxicity in cancer cells in solid tumors.
Rogalska
Aneta
A
Department of Thermobiology, University of Lodz, Banacha 12/16, 90-237 Lodz, Poland. zychan@biol.uni.lodz.pl
Koceva-Chyła
Aneta
A
Jóźwiak
Zofia
Z
D015250:Aclarubicin / Q000494:pharmacology*; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D002452:Cell Count; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008856:Microscopy, Fluorescence; D017382:Reactive Oxygen Species / Q000378:metabolism*
D000903:Antibiotics, Antineoplastic; D017382:Reactive Oxygen Species; D015250:Aclarubicin