Downregulation of NF-kappaB activation in a H4IIE transfectant insensitive to doxorubicin-induced apoptosis.
Cytostatic drugs are administered to cancer patients in order to drive the tumor cells into apoptosis by DNA damage signalling pathway(s). DNA damage also leads to NF-kappaB activation, and it is controversial whether this is exclusively part of a survival process, thus enabling drug resistance, or whether it can also lead to a pro-apoptotic response, thus supporting the therapeutic purpose of drug administration. In the present work, the pathway and outcome of NF-kappaB activation was compared in the doxorubicin sensitive H4IIE rat hepatoma cell and the H4IIE-derived transfectant Yv2-12 which is insensitive to doxorubicin induced apoptosis. In the wild type H4IIE cell, doxorubicin induces serine 536 phosphorylation and nuclear translocation of p65 which however results in reduced rather than increased expression of the anti-apoptotic protein XIAP. Apoptosis in H4IIE cells is accompanied by rapid production of intracellular reactive oxygen species, caspase activation and increased expression of the pro-apoptotic protein Bax. The doxorubicin-insensitive Yv2-12 transfectant differs from its wild type counterpart by the complete failure to activate NF-kappaB in response to doxorubicin. In contrast, serine 536 phosphorylation and nuclear translocation of p65 are even reduced by doxorubicin treatment while the expression of XIAP and Bax remain virtually unchanged. These results show that NF-kappaB activation by doxorubicin in our experimental system proceeds by an atypical pathway resulting in a pro-apoptotic effect and that insensitivity to doxorubicin-induced apoptosis was accompanied by a loss of NF-kappaB activation.
Chovolou
Yvonni
Y
Institute of Toxicology, Heinrich Heine University of Duesseldorf, P.O. Box 10 10 07, D-40001 Düsseldorf, Germany. chovolou@uni-duesseldorf.de
Wätjen
Wim
W
Kampkötter
Andreas
A
Kahl
Regine
R
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects; D004317:Doxorubicin / Q000494:pharmacology*; D008114:Liver Neoplasms, Experimental / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050863:Synaptotagmin I / Q000378:metabolism; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000235:genetics / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D000903:Antibiotics, Antineoplastic; D016328:NF-kappa B; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D050863:Synaptotagmin I; D014409:Tumor Necrosis Factor-alpha; D051636:X-Linked Inhibitor of Apoptosis Protein; D051028:bcl-2-Associated X Protein; D004317:Doxorubicin; D002374:Catalase; D020169:Caspases