Abstract
Although many oncoproteins promote cell growth and proliferation, some also possess the potential to induce cell cycle arrest or cell death by apoptosis. Elevated and deregulated expression of the Myc protein promotes apoptosis in both cultured cells and in some tissues in vivo. Here we describe techniques to detect Myc-induced apoptosis in vitro using flow cytometry, microscopy, and immunoblotting, and in vivo using immunohistochemical staining, immunoblotting, and analysis of RNA expression.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
McMahon SB (2014) MYC and the control of apoptosis. Cold Spring Harb Perspect Med 4:a014407. https://doi.org/10.1101/cshperspect.a014407
Lowe SW, Cepero E, Evan G (2004) Intrinsic tumour suppression. Nature 432:307–315
Pelengaris S, Khan M, Evan GI (2002) Suppression of Myc-induced apoptosis in beta cells exposes multiple oncogenic properties of Myc and triggers carcinogenic progression. Cell 109:321–334
Dansen TB, Whitfield J, Rostker F et al (2006) Specific requirement for Bax, not Bak, in Myc-induced apoptosis and tumor suppression in vivo. J Biol Chem 281:10890–10895
Murphy DJ, Junttila MR, Pouyet L et al (2008) Distinct thresholds govern Myc’s biological output in vivo. Cancer Cell 14:447–457
Li X, Zhang XA, Li X, Xie W, Huang S (2015) MYC-mediated synthetic lethality for treating tumors. Curr Cancer Drug Targets 15:99–115
Cermelli S, Jang IS, Bernard B, Grandori C (2014) Synthetic lethal screens as a means to understand and treat MYC-driven cancers. Cold Spring Harb Perspect Med 4:a014209. https://doi.org/10.1101/cshperspect.a014209
Evan GI, Wyllie AH, Gilbert CS et al (1992) Induction of apoptosis in fibroblasts by c-myc protein. Cell 69:119–128
Littlewood TD, Hancock DC, Danielian PS et al (1995) A modified oestrogen receptor ligand-binding domain as an improved switch for the regulation of heterologous proteins. Nucleic Acids Res 23:1686–1690
Soucek L, Whitfield JR, Sodir NM, Masso-Valles D, Serrano E, Karnezis AN, Swigart LB, Evan GI (2013) Inhibition of Myc family proteins eradicates KRas-driven lung cancer in mice. Genes Dev 27:504–513
Kortlever RM, Sodir NM, Wilson CH, Burkhart DL, Pellegrinet L, Brown Swigart L, Littlewood TD, Evan GI (2017) Myc cooperates with Ras by programming inflammation and immune suppression. Cell 171:1301–1315
Sodir NM, Kortlever RM, Barthet VJA, Campos T, Pellegrinet L, Kupczak S, Anastasiou P, Brown Swigart L, Soucek L, Arends MJ, Littlewood TD, Evan GI (2020) Myc instructs and maintains the pancreatic adenocarcinoma phenotype. Cancer Discov 10(4):588–607. https://doi.org/10.1158/2159-8290.CD-19-0435
Iaccarino I, Hancock D, Evan G et al (2003) c-Myc induces cytochrome c release in Rat1 fibroblasts by increasing outer mitochondrial membrane permeability in a Bid-dependent manner. Cell Death Differ 10:599–608
Juin P, Hunt A, Littlewood T et al (2002) c-Myc functionally cooperates with Bax to induce apoptosis. Mol Cell Biol 22:6158–6169
Wagner AJ, Kokontis JM, Hay N (1994) Myc-mediated apoptosis requires wild-type p53 in a manner independent of cell cycle arrest and the ability of p53 to induce p21waf1/cip1. Genes Dev 8:2817–2830
Zindy F, Eischen CM, Randle DH et al (1998) Myc signaling via the ARF tumor suppressor regulates p53-dependent apoptosis and immortalization. Genes Dev 12:2424–2433
Hermeking H, Eick D (1994) Mediation of c-Myc-induced apoptosis by p53. Science 265:2091–2093
Hoffman B, Liebermann DA (2008) Apoptotic signaling by c-MYC. Oncogene 27:6462–64672
Muthalagu N, Junttila MR, Wiese KE, Wolf E, Morton J, Bauer B, Evan GI, Eilers M, Murphy DJ (2014) BIM is the primary mediator of MYC-induced apoptosis in multiple solid tissues. Cell Rep 8:1347–1353
Hueber A, Zörnig M, Lyon D et al (1997) Requirement for the CD95 receptor-ligand pathway in c-Myc-induced apoptosis. Science 278:1305–1309
Tanaka H, Matsumura I, Ezoe S et al (2002) E2F1 and c-Myc potentiate apoptosis through inhibition of NF-kappaB activity that facilitates MnSOD-mediated ROS elimination. Mol Cell 9:1017–1029
Kauffmann-Zeh A, Rodriguez-Viciana P, Ulrich E et al (1997) Suppression of c-Myc-induced apoptosis by Ras signalling through PI(3)K and PKB. Nature 385:544–548
Juin P, Hueber AO, Littlewood T et al (1999) c-Myc-induced sensitization to apoptosis is mediated through cytochrome c release. Genes Dev 13:1367–1381
Kreuzaler P, Clarke MA, Brown EJ, Wilson CH, Kortlever RM, Piterman N, Littlewood T, Evan GI, Fisher J (2019) Heterogeneity of Myc expression in breast cancer exposes pharmacological vulnerabilities revealed through executable mechanistic modeling. Proc Natl Acad Sci U S A 116:22399–22408
Acknowledgments
This work was supported by Cancer Research UK. We are indebted to our colleagues for advice on the methods presented in this chapter.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Lu, D., Wilson, C., Littlewood, T.D. (2021). Methods for Determining Myc-Induced Apoptosis. In: Soucek, L., Whitfield, J. (eds) The Myc Gene. Methods in Molecular Biology, vol 2318. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1476-1_10
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1476-1_10
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1475-4
Online ISBN: 978-1-0716-1476-1
eBook Packages: Springer Protocols