Increased activity and sensitivity of mitochondrial respiratory enzymes to tumor necrosis factor alpha-mediated inhibition is associated with increased cytotoxicity in drug-resistant leukemic cell lines.
The drug-resistant leukemic cell lines, CEM/VLB100 and K/DAU600, are more sensitive to tumor necrosis factor alpha (TNFalpha)-mediated cytotoxicity compared with their parental cell lines, CCRF-CEM and K562 cl.6. Drug-resistant leukemic cell lines have more active mitochondrial function, which is associated with a greater susceptibility to TNFalpha-induced respiratory inhibition. TNFalpha blocked electron transfer at three sites, NADH dehydrogenase (complex I), succinate dehydrogenase (complex II), and cytochrome c oxidase (complex IV). Respiratory rate and electron transport chain enzyme activities were significantly inhibited in the drug-resistant, TNF-sensitive cell lines. Respiratory inhibition preceded cell death by at least 5 to 8 hours. The respiratory failure was not compensated for by appropriate up-regulation of the glycolytic pathway. Increasing mitochondrial respiratory rate and enzyme activities by long-term culture with 2 mmol/L adenosine 5'-diphosphate (ADP) and Pi sensitized both drug-sensitive and drug-resistant cells to TNFalpha-induced cytolysis. Intramitochondrial free radicals generated by paraquat only had a limited and delayed effect on respiratory inhibition and cytolysis in comparison with the effect of TNFalpha. We conclude that TNFalpha-induced cytotoxicity in leukemic cells is, at least in part, mediated by inhibition of mitochondrial respiration. Free radical generation by TNFalpha may not directly lead to the observed inhibition of the mitochondrial electron transport and other mechanisms must be involved.
Jia
L
L
Department of Haematology, London Hospital Medical College, London, UK.
Kelsey
S M
SM
Grahn
M F
MF
Jiang
X R
XR
Newland
A C
AC
D000244:Adenosine Diphosphate / Q000494:pharmacology; D016923:Cell Death; D002478:Cells, Cultured; D003630:Daunorubicin / Q000494:pharmacology; D003864:Depression, Chemical; D019008:Drug Resistance, Neoplasm; D004579:Electron Transport / Q000187:drug effects*; D042963:Electron Transport Complex II; D003576:Electron Transport Complex IV / Q000037:antagonists & inhibitors*; D004791:Enzyme Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D005609:Free Radicals; D006019:Glycolysis; D006801:Humans; D007938:Leukemia / Q000473:pathology*; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000473:pathology; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000473:pathology; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors*; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors*; D009363:Neoplasm Proteins / Q000032:analysis*; D010088:Oxidoreductases / Q000037:antagonists & inhibitors*; D010269:Paraquat / Q000494:pharmacology; D013385:Succinate Dehydrogenase / Q000037:antagonists & inhibitors*; D013481:Superoxides / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology* / Q000633:toxicity; D014747:Vinblastine / Q000494:pharmacology
D004791:Enzyme Inhibitors; D005609:Free Radicals; D009097:Multienzyme Complexes; D009363:Neoplasm Proteins; D014409:Tumor Necrosis Factor-alpha; D013481:Superoxides; D014747:Vinblastine; D000244:Adenosine Diphosphate; D010088:Oxidoreductases; D042963:Electron Transport Complex II; D013385:Succinate Dehydrogenase; D016660:NAD(P)H Dehydrogenase (Quinone); D003576:Electron Transport Complex IV; D010269:Paraquat; D003630:Daunorubicin