Pretreatment with PKC inhibitor triggers TNF-α induced apoptosis in TNF-α–resistant B16 melanoma BL6 cells
详细信息 查看全文
文摘
Tumor necrosis factor α (TNF-α) modulates various events through several different pathways. Many tumor cells are resistant to this cytokine. Pretreatment of these cells with actinomycin D enhances TNF-α–induced apoptosis. In the present study, we investigated the mechanism of this enhancement and whether or not the apoptosis of TNF-α–resistant cancer cells can be induced by the inhibition of Protein kinase C (PKC). When TNF-α was added after inhibition of PKC by H7, apoptosis was observed, and companied with the activation of nuclear factor kappa B (NF-κB). After the inhibition of protein kinase B (Akt) by LY294002 or p38 mitogen-activated protein kinase (p38MAPK) by SB203580, the addition of TNF-α did not cause apoptosis. However, after the inhibition of MAPK/extracellular signal-regulated kinase kinase 1/2 (MEK1/2) with U0126, apoptosis was observed when TNF-α was added. In the Western blotting analysis, phosphorylation of MEK1/2 occurred at 60 minutes after the addition of TNF-α. However, it was noted that after pretreatment with H7, a significant decrease in phosphorylated MEK1/2 was observed. The present findings suggest that MEK1/2 plays an important role in TNF-α–resistance in TNF-α–resistant B16 melanoma BL6 cells. Furthermore, it was found that MEK1/2 is more important than NF-κB, Akt, and p38MAPK in anti-apoptotic PKC signaling and that TNF-α–resistance can be overcome by inhibiting MEK1/2. These results suggest the possibility of development of a new anticancer drug treatment.