TAILIEUCHUNG - Báo cáo khoa học: Etoposide upregulates Bax-enhancing tumour necrosis factor-related apoptosis inducing ligand-mediated apoptosis in the human hepatocellular carcinoma cell line QGY-7703

Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) has attractedmuch attention because of its ability to kill tumour cells. In this study, we demonstrated that treatment of QGY-7703 cells with the combination of TRAIL and etoposide resulted in synergistic cytotoxic effects. In dissecting the mechanism underlying this syner-gistic effect, we found that treatment with etoposide alone resulted in the upregulation of Bax, while the level of trun-catedBid (tBid)wasunchanged. | Eur. J. Biochem. 270 2721-2731 2003 FEBS 2003 doi Etoposide upregulates Bax-enhancing tumour necrosis factor-related apoptosis inducing ligand-mediated apoptosis in the human hepatocellular carcinoma cell line QGY-7703 Lin Miao Peng Yi Yi Wang and Mian Wu Department of Molecular and Cell Biology Key Laboratory of Structural Biology School of Life Sciences University of Science and Technology of China Hefei Anhui China Tumour necrosis factor-related apoptosis-inducing ligand TRAIL has attracted much attention because of its ability to kill tumour cells. In this study we demonstrated that treatment of QGY-7703 cells with the combination of TRAIL and etoposide resulted in synergistic cytotoxic effects. In dissecting the mechanism underlying this synergistic effect we found that treatment with etoposide alone resulted in the upregulation of Bax while the level of truncated Bid tBid was unchanged. In contrast while treatment with TRAIL alone significantly increased the level of tBid the expression of Bax remained unaffected. The enhanced apoptosis was accompanied by an increased release of cytochrome c and second mitochondria-derived activator of caspase direct IAP binding protein with low pI DIABLO from mitochondria leading to the activation of cellular caspase-8 -9 -3 and -7 as well as poly ADP-ribose polymerase. This enhanced release of cytochrome c and second mitochondria-derived activator of caspase DIABLO was inhibited by the general caspase inhibitor N-benzyloxycar-bonyl-Val-Ala-Asp-fluoromethylketone. The RT-PCR and Western blotting results demonstrated that the levels of both mRNA and protein for death receptor-4 death receptor-5 and decoy receptor-2 remained unchanged in response to etoposide indicating that the synergistic effect of TRAIL and etoposide is not a result of increasing the expression for TRAIL receptors but rather is associated with amplification of the mitochondrial signal pathway. Keywords p53 Bax tBid .

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