Abstract
The human adenovirus E4orf4 protein, when expressed alone, induces p53-independent death in a wide range of cancer cells. Earlier studies by our groups suggested that although in some cases cell death can be associated with some hallmarks of apoptosis, it is not always affected by caspase inhibitors. Thus it is unlikely that E4orf4-induced cell death occurs uniquely through apoptosis. In the present studies using H1299 human lung carcinoma cells as a model system we found that death is induced in the absence of activation of any of the caspases tested, accumulation of reactive oxygen species, or release of cytochrome c from mitochondria. E4orf4 caused a substantial change in cell morphology, including vigorous membrane blebbing, multiple nuclei in many cells and increased cell volume. Most of these characteristics are not typical of apoptosis, but they are of necrosis. FACS analysis and western blotting for cell cycle markers showed that E4orf4-expressing cells became arrested in G2/M and also accumulated high levels of cyclin E. The presence of significant numbers of tetraploid and polyploid cells and some cells with micronuclei suggested that E4orf4 appears to induce death in these cells through a process resulting from mitotic catastrophe.
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Acknowledgements
We thank Dennis Takayesu for technical support and Denis Paquette, Helen Chan and Mark Watson from Gemin X Biotechnologies Inc. for making some of the DNA constructs and for help in some experimental protocols. We also thank Eric Massicotte and Nathalie Tessier of Institut des reserches cliniques de Montréal and Ken McDonald at McGill Life Sciences Complex for performing flow cytometry analyses, and Jacynthe Lalibeté from the Department of Pharmacology and Therapeutics at McGill University for confocal microscopy study. Gordon Shore provided anti-caspase and anti-actin antibodies. This work was supported by grants from the Canadian Cancer Society through the National Cancer Institute of Canada (PEB, JNL), the Canadian Institutes of Health Research (PEB, JNL), and from Gemin X Biotechnologies Inc (PEB).
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Li, S., Szymborski, A., Miron, MJ. et al. The adenovirus E4orf4 protein induces growth arrest and mitotic catastrophe in H1299 human lung carcinoma cells. Oncogene 28, 390–400 (2009). https://doi.org/10.1038/onc.2008.393
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DOI: https://doi.org/10.1038/onc.2008.393
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