Abstract
Expressing oncogenes in normal somatic human cells leads to cellular senescence after just a few cell division cycles. In cells that are more resistant to culture stresses, such as human dermal fibroblasts, this oncogene-induced senescence (OIS) is a result of a DNA damage response (DDR) that is activated due to the formation of DNA lesions at both non-telomeric and telomeric DNA sequences. DNA lesions can be visualized as DDR foci by immunofluorescence microscopy using antibodies against a number of DDR factors, including ϒ-H2AX and 53BP1. Over time and as cells remain arrested in OIS, non-telomeric DDR foci progressively become resolved, while telomeric DDR foci, also called dysfunctional telomeres, persist. Here we describe a protocol to detect dysfunctional telomeres in cultured human cells, to monitor a temporal enrichment of dysfunctional telomeres in cells that had undergone OIS, and to detect dysfunctional telomeres in paraffin-embedded and formalin-fixed human tissue.
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Acknowledgments
Research in the laboratory of UH is supported by the National Cancer Institute of the NIH (R01CA136533, R01CA184572). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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Patel, P.L., Herbig, U. (2017). Detection of Dysfunctional Telomeres in Oncogene-Induced Senescence. In: Nikiforov, M. (eds) Oncogene-Induced Senescence. Methods in Molecular Biology, vol 1534. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6670-7_6
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DOI: https://doi.org/10.1007/978-1-4939-6670-7_6
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-6670-7
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