Mechanism of cytotoxic action of perfluorinated acids II. Disruption of mitochondrial bioenergetics.
PFAs and derivatives due to perfect technological properties are broadly applied in industry and consumer goods, and in consequence widely disseminated, environmentally bioaccumulative and found at ppb level in human serum. Earlier we revealed that in vitro cytotoxicity increases with chain length (CF(6)-CF(14)). The compounds dissipate plasma membrane potential and acidify of cytosol. Here we determine whether there is an association between the protonophoric uncoupling of respiration and disruption of bioenergetics caused by CF(6)-CF(12) on HCT116 cell apoptosis. Again the effects were stronger for longer molecules. Incubation of cells with CF(10) stimulated time-dependent generation of reactive oxygen species, opening of mitochondrial permeability transition (MPT) pore, release of cytochrome c, activation of caspases and depletion of intracellular level of ATP occurring in intrinsic pathway of apoptosis. Incubation with decanoic acid (DA) did not lead to mitochondrial dysfunctions neither to cell cycle disturbances. Synchronized removal of the phosphorylated state of Akt, ERK1/2 and PKCdelta/theta kinases by CF(10) suggests presence of concerted action to uninhibit Bad protein activation and a cascade of intrinsic pathway of apoptosis. Blocking MPT pore by cyclosporin A (CsA) led to a reduction of mitochondrial potential dissipation (mtDeltaPsi). Such cells neither showed cytochrome c release nor the downstream activation of caspase-9 and caspase-3. Our results confirm that mitochondria play a crucial role in perfluorochemicals induced apoptosis by releasing apoptotic signals through MPT pore.
Kleszczyński
Konrad
K
Intercollegiate Faculty of Biotechnology, Medical University of Gdańsk, Department of Molecular Enzymology ul. Debinki 1, 80-211 Gdańsk, Poland.
Stepnowski
Piotr
P
Składanowski
Andrzej C
AC
D000227:Adenine Nucleotides / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D045304:Cytochromes c / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D004734:Energy Metabolism / Q000187:drug effects*; D005466:Fluorocarbons / Q000633:toxicity*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007202:Indicators and Reagents; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008856:Microscopy, Fluorescence; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism*; D010101:Oxygen Consumption / Q000187:drug effects; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
D000227:Adenine Nucleotides; D005466:Fluorocarbons; D007202:Indicators and Reagents; D017382:Reactive Oxygen Species; D000255:Adenosine Triphosphate; D045304:Cytochromes c; D011494:Protein Kinases; D020169:Caspases