TNF-alpha, major pro-inflammatory cytokine, is involved in physiological and pathophysiological processes in the skeletal muscle. For example, NF-a modulates myogenesis by affecting directly myoblast cell Proliferation/differentiation or induction of apoptosis. We investigated the effects of TNF-alpha on transcriptional changes occurring during myoblast differentiation in C2C 12 cells, a well-characterized in vitro model of myogenesis. The cells were cultured in media to allow for proliferation for 24 hrs followed by differentiation media for 2, 24 and 48 hrs in the presence or absence of recombinant mouse TNF-alpha. The microarray analysis included 715 genes and was conducted in triplicate. Only genes showing a differential expression (both increased and decreased) 1.7-fold or greater (p < 0.05) than control cells were analyzed. Clustering analysis based on expression profiles and gene function revealed sets of genes differentially regulated between proliferating and differentiating C2CI2 cells. TNF-alpha induced increased expression of genes coding mediators of apoptosis, cell-matrix interactions, proteolysis and C-C chemokines. The expression of selected genes was validated by real time RT -PCR, for both the direction and magnitude of the gene expression observed by the cDNA array. This study supports the hypothesis that a complex regulatory role of TNF-alpha exits in skeletal myogenesis.
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