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B mediates the protein loss induced by TNF-
in
differentiated skeletal muscle myotubes
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030
Nuclear factor-
B (NF-B)
regulates the transcription of a variety of genes involved in immune
responses, cell growth, and cell death. However, the role of NF-B in
muscle biology is poorly understood. We recently reported that tumor
necrosis factor-
(TNF-) rapidly activates NF-B in
differentiated skeletal muscle myotubes and that TNF- acts directly
on the muscle cell to induce protein degradation. In the present study,
we ask whether NF-B mediates the protein loss induced by TNF-. We
addressed this problem by creating stable, transdominant negative
muscle cell lines. C2C12 myoblasts were transfected with viral plasmid
constructs that induce overexpression of mutant I-B proteins that
are insensitive to degradation via the ubiquitin-proteasome pathway.
These mutant proteins selectively inhibit NF-B activation. We found
that differentiated myotubes transfected with the empty viral vector
(controls) underwent a drop in total protein content and in fast-type
myosin heavy-chain content during 72 h of exposure to TNF-. In
contrast, total protein and fast-type myosin heavy-chain levels were
unaltered by TNF- in the transdominant negative cell lines. TNF-
did not induce apoptosis in any cell line, as assessed by DNA ladder
and annexin V assays. These data indicate that NF-B is an essential
mediator of TNF--induced catabolism in differentiated muscle cells.
cachexia; cytokine; free radicals; signal transduction; inflammation
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