Abstract
The p53 tumor suppressor protein plays a central role in mediating the cellular response to a variety of stresses. Activation of p53 signaling will trigger cell cycle arrest or apoptosis in normal cells, depending on such factors as cell type and genetic context. The ability of a cell to circumvent either of these p53-directed outcomes leads to inappropriate proliferation, thereby contributing to the development of cancer. As such, tumors frequently escape the apoptotic pathway in response to cell stress. DNA-damaging agents, however, achieve significant tumor cytotoxicity in spite of this hallmark characteristic. Tumors treated with DNA-damaging drugs often undergo alternate forms of cell death, such as senescence or mitotic catastrophe, in addition to apoptosis that may ultimately lead to regression. Although not a predictor of chemotherapy response in patients per se, p53 status in tumor-derived cells is frequently a determinant of the death pathway promoted by these agents. The cytotoxic effects of DNA-damaging agents can be readily appreciated using such tools as cell cycle analysis, phopsho-H3Ser10 immunoblotting, and annexin V detection.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Stewart ZA, Pietenpol JA (2001) p53 Signaling and cell cycle checkpoints. Chem Res Toxicol 14:243–263
Vousden KH, Lu X (2002) Live or let die: the cell’s response to p53. Nat Rev Cancer 2:594–604
Sansom OJ, Clarke AR (2000) P53 null mice: damaging the hypothesis? Mutat Res 452:149–162
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100:57–70
Lowe SW, Ruley HE, Jacks T, Housman DE (1993) p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell 74:957–967
Lowe SW, Schmitt EM, Smith SW, Osborne BA, Jacks T (1993) p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature 362:847–849
Gudkov AV, Komarova EA (2003) The role of p53 in determining sensitivity to radiotherapy. Nat Rev Cancer 3:117–129
Merritt AJ, Allen TD, Potten CS, Hickman JA (1997) Apoptosis in small intestinal epithelial from p53-null mice: evidence for a delayed, p53-independent G2/M-associated cell death after gamma-irradiation. Oncogene 14:2759–2766
Han JW, Dionne CA, Kedersha NL, Goldmacher VS (1997) p53 status affects the rate of the onset but not the overall extent of doxorubicin-induced cell death in rat-1 fibroblasts constitutively expressing c-Myc. Cancer Res 57:176–182
Tannock IF, Lee C (2001) Evidence against apoptosis as a major mechanism for reproductive cell death following treatment of cell lines with anti-cancer drugs. Br J Cancer 84:100–105
Kemp CJ, Sun S, Gurley KE (2001) p53 induction and apoptosis in response to radio- and chemotherapy in vivo is tumor-type-dependent. Cancer Res 61:327–332
Agarwal ML, Agarwal A, Taylor WR, Stark GR (1995) p53 controls both the G2/M and the G1 cell cycle checkpoints and mediates reversible growth arrest in human fibroblasts. Proc Natl Acad Sci USA 92:8493–8497
Flatt PM, Tang LJ, Scatena CD, Szak ST, Pietenpol JA (2000) p53 regulation of G(2) checkpoint is retinoblastoma protein dependent. Mol Cell Biol 20:4210–4223
Meulmeester E, Jochemsen AG (2008) p53: a guide to apoptosis. Curr Cancer Drug Targets 8:87–97
Schmitt CA, Rosenthal CT, Lowe SW (2000) Genetic analysis of chemoresistance in primary murine lymphomas. Nat Med 6:1029–1035
Gewirtz DA, Holt SE, Elmore LW (2008) Accelerated senescence: an emerging role in tumor cell response to chemotherapy and radiation. Biochem Pharmacol 76:947–957
Dimri GP (2005) What has senescence got to do with cancer? Cancer Cell 7:505–512
Xue W, Zender L, Miething C, Dickins RA, Hernando E, Krizhanovsky V, Cordon-Cardo C, Lowe SW (2007) Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas. Nature 445:656–660
Brown JM, Wilson G (2003) Apoptosis genes and resistance to cancer therapy: what does the experimental and clinical data tell us? Cancer Biol Ther 2:477–490
Bunz F, Dutriaux A, Lengauer C, Waldman T, Zhou S, Brown JP, Sedivy JM, Kinzler KW, Vogelstein B (1998) Requirement for p53 and p21 to sustain G2 arrest after DNA damage. Science 282:1497–1501
Castedo M, Perfettini JL, Roumier T, Andreau K, Medema R, Kroemer G (2004) Cell death by mitotic catastrophe: a molecular definition. Oncogene 23:2825–2837
Mansilla S, Bataller M, Portugal J (2006) Mitotic catastrophe as a consequence of chemotherapy. Anticancer Agents Med Chem 6:589–602
Mansilla S, Priebe W, Portugal J (2006) Mitotic catastrophe results in cell death by caspase-dependent and caspase-independent mechanisms. Cell Cycle 5:53–60
Nowak SJ, Corces VG (2004) Phosphorylation of histone H3: a balancing act between chromosome condensation and transcriptional activation. Trends Genet 20:214–220
Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MH (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415–1420
DeWeese TL, Walsh JC, Dillehay LE, Kessis TD, Hedrick L, Cho KR, Nelson WG (1997) Human papillomavirus E6 and E7 oncoproteins alter cell cycle progression but not radiosensitivity of carcinoma cells treated with low-dose-rate radiation. Int J Radiat Oncol Biol Phys 37:145–154
Portugal J, Bataller M, Mansilla S (2009) Cell death pathways in response to antitumor therapy. Tumori 95:409–421
Bunz F, Hwang PM, Torrance C, Waldman T, Zhang Y, Dillehay L, Williams J, Lengauer C, Kinzler KW, Vogelstein B (1999) Disruption of p53 in human cancer cells alters the responses to therapeutic agents. J Clin Invest 104:263–269
Acknowledgments
The authors wish to thank Jerry Chipuk and Kostas Floros for guidance with Annexin V staining and detection protocols. The authors are supported by grants from the National Cancer Institute.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Senturk, E., Manfredi, J.J. (2013). Determine the Effect of p53 on Chemosensitivity. In: Deb, S., Deb, S. (eds) p53 Protocols. Methods in Molecular Biology, vol 962. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-236-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-62703-236-0_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-235-3
Online ISBN: 978-1-62703-236-0
eBook Packages: Springer Protocols