Non-apoptotic function of caspases in a cellular model of hydrogen peroxide-associated colitis

作者：Poehlmann Angela^*; Reissig Kathrin; Just Andrea; Walluscheck Diana; Hartig Roland; Schinlauer Antje; Lessel Wiebke; Guenther Thomas; Silver Andrew; Steinberg Pablo; Roessner Albert

来源：Journal of Cellular and Molecular Medicine, 2013, 17(7): 901-913.

DOI：10.1111/jcmm.12079

• hydrogen peroxide-associated colitis
• DNA-damage checkpoints
• non-apoptotic caspase function
• JNK-dependent cell cycle arrests
• inflammation
• neoplastic transformation
• ATM degradation
• gamma-H2AX

Oxidative stress, caused by reactive oxygen species (ROS), is a major contributor to inflammatory bowel disease (IBD)-associated neoplasia. We mimicked ROS exposure of the epithelium in IBD using non-tumour human colonic epithelial cells (HCEC) and hydrogen peroxide (H2O2). A population of HCEC survived H2O2-induced oxidative stress via JNK-dependent cell cycle arrests. Caspases, p21(WAF1) and gamma-H2AX were identified as JNK-regulated proteins. Up-regulation of caspases was linked to cell survival and not, as expected, to apoptosis. Inhibition using the pan-caspase inhibitor Z-VAD-FMK caused up-regulation of gamma-H2AX, a DNA-damage sensor, indicating its negative regulation via caspases. Cell cycle analysis revealed an accumulation of HCEC in the G(1)-phase as first response to oxidative stress and increased S-phase population and then apoptosis as second response following caspase inhibition. (1)- and S-phase by overriding the G(1)/S- and intra-S checkpoints despite DNA-damage. (2)/M-phase and decreased apoptosis. Caspases mediate survival of oxidatively damaged HCEC via gamma-H2AX suppression, although its direct proteolytic inactivation was excluded. Conversely, we found that oxidative stress led to caspase-dependent proteolytic degradation of the DNA-damage checkpoint protein A that is upstream of gamma-H2AX. As a consequence, undetected DNA-damage and increased proliferation were found in repeatedly H2O2-exposed HCEC. Such features have been associated with neoplastic transformation and appear here to be mediated by a non-apoptotic function of caspases. Overexpression of upstream p-JNK in active ulcerative colitis also suggests a potential importance of this pathway in vivo.

• 出版日期2013-7