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B Activation and Reactive Oxygen Intermediates
Cytokine Research Laboratory, Department of Molecular Oncology, University of Texas M. D. Anderson Cancer Center, Houston, TX 77030
LPS, a component of the cell wall in Gram-negative bacteria,
induces inflammation and septic shock syndrome by stimulating various
inflammatory cytokines including TNF. How LPS affects the TNF-mediated
cellular responses, however, is not understood. In this study, the
effect of LPS on TNF-mediated apoptosis in human histiocytic lymphoma
U-937 cells was investigated. We found that treatment of cells with LPS
completely abolished TNF-mediated cytotoxicity and activation of
caspase-3. LPS-chelating antibiotic, polymyxin B, suppressed the
antiapoptotic activity, indicating the specificity of the effect.
Within minutes, LPS through CD14 induced the activation of NF-
B,
degradation of IB
(inhibitory subunit of NF-B) and IBß,
and nuclear translocation of p65. An antioxidant, pyrrolidine
dithiocarbamate, which blocked LPS-induced NF-B activation, also
abolished the antiapoptotic effects of LPS at the same time. Besides
TNF, the apoptosis induced by taxol and okadaic acid was also sensitive
to LPS-induced NF-B activation, whereas that induced by
H2O2, doxorubicin, daunomycin, vincristine, and
vinblastine was NF-B insensitive. Tumor cells that constitutively
expressed NF-B also showed resistance to the apoptotic effects of
TNF, taxol, and okadaic acid, but sensitivity to all other agents,
indicating the critical role of NF-B in blocking apoptosis induced
by certain agents. Overall, these results indicate that LPS induces
resistance to the apoptotic effects of TNF and other agents, and that
NF-B activation, whether induced or constitutive, inhibits this
apoptosis.
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