Goniothalamin-induced oxidative stress, DNA damage and apoptosis via caspase-2 independent and Bcl-2 independent pathways in Jurkat T-cells

Salmaan Hussain Inayat Hussain, K M Chan, N F Rajab, L B Din, Chow Sek Chuen, A Kizilors, F Farzaneh, G T Williams

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    Abstract

    Goniothalamin (GTN) isolated from Goniothalamus sp. has been demonstrated to induce apoptosis in a variety of cancer cell lines including Jurkat T leukemia cells However, the mechanism of GTN-induced apoptosis upstream of mitochondria is still poorly defined in this study, GTN caused a decrease in GSH with an elevation of reactive oxygen species as early as 30 min and DNA damage as assessed by Comet assay Analysis using topoisomerase 11 processing of supercoiled pBR 322 DNA showed that GTN caused DNA damage via a topoisomerase II-independent pathway suggesting that cellular oxidative stress may contribute to genotoxicity. A 12-fold increase of caspase-2 activity was observed in GTN-treated Jurkat cells after 4 h treatment and this was confirmed using Western blotting Although the caspase-2 inhibitor Z-VDVAD-FMK inhibited the proteolytic activity of caspase-2, apoptosis ensued confirming that caspase-2 activity was not crucial for GTN-induced apoptosis However, GTN-induced apoptosis was completely abrogated by N-acetylcysteine further confirming the role of oxidative stress Since cytochrome c release was observed as early as 1 h without any appreciable change in Bcl-2 protein expression, we further investigated whether overexpression of Bcl-2 confers resistance in GTN-induced cytotoxicity Using a panel of Jurkat Bcl-2 transfectants, GTN cytotoxicity was not abrogated in these cells. In conclusion, GTN induces DNA damage and oxidative stress resulting in apoptosis which is independent of both caspase-2 and Bcl-2. (C) 2009 Elsevier Ireland Ltd All rights reserved
    Original languageEnglish
    Pages (from-to)108 - 114
    Number of pages7
    JournalToxicology Letters
    Volume193
    Issue number1
    DOIs
    Publication statusPublished - 2010

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