Effects of permeability transition inhibition and decrease in cytochrome c content on doxorubicin toxicity in K562 cells.
As mitochondria play a key role in the commitment to cell death, we have investigated the mitochondrial consequences of resistance to doxorubicin (DOX) in K562 cells. We found that the permeability transition pore (PTP) inhibitor cyclosporine A (CsA) failed to inhibit PTP opening in the resistant clone. Moreover, the Ca2+ loading capacity in the resistant clone was identical to that observed in the parent cells in the presence of CsA, suggesting that the PTP was already inhibited in a CsA-like manner in the resistant cells. In agreement with this proposal, the mitochondrial target of CsA cyclophilin D (CyD) decreased by half in the resistant cells. The levels of adenine nucleotide translocator, voltage anion-dependent channel, Bax, Bcl-2, Bcl-xL, AIF and Smac/Diablo, were similar in both cell lines, whereas cytochrome c content was divided by three in the resistant cells. Since P-glycoprotein inhibition did not restore DOX toxicity in the resistant cells, while DOX-induced cell death in the parent cells was prevented by either PTP inhibition or siRNA-induced decrease in cytochrome c content, we conclude that the inhibition of PTP opening and the decrease in cytochrome c content participate in the mechanism that makes K562 cells resistant to DOX.
De Oliveira
F
F
INSERM E-0221 Bioénergétique Fondamentale et Appliquée, Université Joseph Fourier, Grenoble, France.
Chauvin
C
C
Ronot
X
X
Mousseau
M
M
Leverve
X
X
Fontaine
E
E
D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D017209:Apoptosis; D015153:Blotting, Western; D045304:Cytochromes c / Q000378:metabolism*; D004317:Doxorubicin / Q000633:toxicity*; D019008:Drug Resistance, Neoplasm; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism*; D020014:K562 Cells / Q000187:drug effects / Q000648:ultrastructure; D008564:Membrane Potentials; D033681:Mitochondrial Membrane Transport Proteins / Q000037:antagonists & inhibitors*; D000083162:Mitochondrial Permeability Transition Pore; D034741:RNA, Small Interfering / Q000235:genetics
D000903:Antibiotics, Antineoplastic; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D034741:RNA, Small Interfering; D004317:Doxorubicin; D045304:Cytochromes c