Abstract
A major role for c-Myc in the proliferation of normal cells is attributed to its ability to promote progression through G1 and into S phase of the cell cycle. The absolute requirement of c-Myc for cell cycle progression in human tumor cells has not been comprehensively addressed. In the present work, we used a lentiviral-based short hairpin RNA (shRNA) expression vector to stably reduce c-Myc expression in a large number of human tumor cell lines and in three different types of normal human cells. In all cases, cell proliferation was severely inhibited, with normal cells ultimately undergoing G0/G1 growth arrest. In contrast, tumor cells demonstrated a much more variable cell cycle response with cells from several lines accumulating in S or G2/M phases. Moreover, in some tumor lines, the phase of cell cycle arrest caused by inhibition of c-Myc could be altered by depleting tumor suppressor protein p53 or its transcriptional target p21CIP/WAF. Our data suggest that, as in the case of normal cells, c-Myc is essential for sustaining proliferation of human tumor cells. However its rate-limiting role in cell cycle control is variable and is reliant upon the status of other cell cycle regulators.
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References
Adhikary S, Eilers M . (2005). Transcriptional regulation and transformation by Myc proteins. Nat Rev Mol Cell Biol 6: 635–645.
Agarwal ML, Agarwal A, Taylor WR, Chernova O, Shama Y, Stark GR . (1998). A p53-dependent S-phase checkpoint helps to protect cells from DNA damage in response to starvation for pyrimidine nucleotides. Proc Natl Acad Sci USA 95: 14775–14780.
Arvanitis C, Felsher DW . (2006). Conditional transgenic models define how MYC initiates and maintains tumorigenesis. Semin Cancer Biol 16: 313–317.
Athar M, Tang X, Lee JL, Kopelovich L, Kim AL . (2006). Hedgehog signalling in skin development and cancer. Exp Dermatol 15: 667–677.
Baena E, Gandarillas A, Vallespinos M, Zanet J, Bachs O, Redondo C et al. (2005). c-Myc regulates cell size and ploidy but is not essential for postnatal proliferation in live. Proc Natl Acad Sci USA 102: 7286–7291.
Bernard S, Eilers M . (2006). Control of cell proliferation and growth by Myc proteins. Results Probl Cell Differ 42: 329–342.
Bettess MD, Dubois N, Murphy MJ, Dubey C, Roger C, Robine S et al. (2005). c-Myc is required for the formation of intestinal crypts but dispensable for homeostasis of the adult intestinal epithelium. Mol Cell Biol 25: 7868–7878.
Boiko AD, Porteous S, Razorenova OV, Krivokrysenko VI, Williams BR, Gudkov AV et al. (2006). A systematic search for downstream mediators of tumor suppressor function of p53 reveals a major role of BTG2 in suppression of Ras-induced transformation. Genes Dev 20: 236–252.
Boxer RB, Jang JW, Sintasath L, Chodosh LA . (2004). Lack of sustained regression of c-MYC-induced mammary adenocarcinomas following brief or prolonged MYC inactivation. Cancer Cell 6: 577–586.
Chen JP, Lin C, Xu CP, Zhang XY, Fu M, Deng YP . (2001). Molecular therapy with recombinant antisense c-myc adenovirus for human gastric carcinoma cells in vitro and in vivo. J Gastroenterol Hepatol 6: 22–28.
Citro G, D'Agnano I, Leonetti C, Perini R, Bucci B, Zon G et al. (1998). c-myc antisense oligodeoxynucleotides enhance the efficacy of cisplatin in melanoma chemotherapy in vitro and in nude mice. Cancer Res 58: 283–289.
D'Agnano I, Valentini A, Fornari C, Bucci B, Starace G, Felsani A et al. (2001). Myc down-regulation induces apoptosis in M14 melanoma cells by increasing p27(kip1) levels. Oncogene 20: 2814–2825.
Dang CV . (1999). c-Myc target genes involved in cell growth, apoptosis, and metabolism. Mol Cell Biol 19: 1–11.
Dang CV, O'Donnell KA, Zeller KI, Nguyen T, Osthus RC, Li F . (2006). The c-Myc target gene network. Semin Cancer Biol 16: 253–264.
de Alboran IM, O'Hagan RC, Gartner F, Malynn B, Davidson L, Rickert R et al. (2001). Analysis of C-MYC function in normal cells via conditional gene-targeted mutation. Immunity 14: 45–55.
Denoyelle C, Abou-Rjaily G, Bezrookove V, Verhaegen M, Johnson TM, Fullen DR et al. (2006). Anti-oncogenic role of the endoplasmic reticulum differentially activated by mutations in the MAPK pathway. Nat Cell Biol 8: 1053–1063.
Eisenman RN . (2001). Deconstructing myc. Genes Dev 15: 2023–2030.
El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM et al. (1993). WAF1, a potential mediator of p53 tumor suppression. Cell 75: 817–825.
Felsher DW, Bishop JM . (1999). Reversible tumorigenesis by MYC in hematopoietic lineages. Mol Cell 4: 199–207.
Fernandez Y, Verhaegen M, Miller TP, Rush JL, Steiner P, Opipari AW et al. (2005). Differential regulation of noxa in normal melanocytes and melanoma cells by proteasome inhibition: therapeutic implications. Cancer Res 65: 6294–6304.
Grandori C, Cowley SM, James LP, Eisenman RN . (2000). The Myc/Max/Mad network and the transcriptional control of cell behavior. Annu Rev Cell Dev Biol 16: 653–659.
Gottifredi V, Prives C . (2005). The S phase checkpoint: when the crowd meets at the fork. Semin Cell Dev Biol 16: 355–368.
Hahn WC, Weinberg RA . (2002). Rules for making human tumor cells. N Engl J Med 347: 1593–1603.
Hanahan D, Weinberg RA . (2000). The hallmarks of cancer. Cell 100: 57–70.
Hermeking H, Rago C, Schuhmacher M, Li Q, Barrett JF, Obaya AJ et al. (2000). Identification of CDK4 as a target of c-Myc. Proc Natl Acad Sci USA 97: 2229–2234.
Hermeking H . (2003). The MYC oncogene as a cancer drug target. Curr Cancer Drug Targets 3: 163–175.
Ivanova N, Dobrin R, Lu R, Kotenko I, Levorse J, DeCoste C et al. (2006). Dissecting self-renewal in stem cells with RNA interference. Nature 442: 533–538.
Jain M, Arvanitis C, Chu K, Dewey W, Leonhardt E, Trinh M et al. (2002). Sustained loss of a neoplastic phenotype by brief inactivation of MYC. Science 297: 102–104.
Li F, Xiang Y, Potter J, Dinavahi R, Dang CV, Lee LA . (2006). Conditional deletion of c-myc does not impair liver regeneration. Cancer Res 66: 5608–5612.
Lutz W, Leon J, Eilers M . (2002). Contributions of Myc to tumorigenesis. Biochim Biophys Acta 1602: 61–71.
Malynn BA, de Alboran IM, O'Hagan RC, Bronson R, Davidson L, DePinho RA et al. (2000). N-myc can functionally replace c-myc in murine development, cellular growth, and differentiation. Genes Dev 14: 1390–1399.
Mateyak MK, Obaya AJ, Adachi S, Sedivy JM . (1997). Phenotypes of c-Myc-deficient rat fibroblasts isolated by targeted homologous recombination. Cell Growth Differ 8: 1039–1048.
McGuffie EM, Pacheco D, Carbone GM, Catapano CV . (2000). Antigene and antiproliferative effects of a c-myc-targeting phosphorothioate triple helix-forming oligonucleotide in human leukemia cells. Cancer Res 60: 3790–3799.
Meyer N, Kim SS, Penn LZ . (2006). The Oscar-worthy role of Myc in apoptosis. Semin Cancer Biol 16: 275–277.
Nesbit CE, Tersak JM, Prochownik EV . (1999). Myc oncogenes and human neoplastic disease. Oncogene 18: 3004–3016.
Nikiforov MA, Chandriani S, O'Connell B, Petrenko O, Kotenko I, Beavis A et al. (2002). A functional screen for Myc-responsive genes reveals serine hydroxymethyltransferase, a major source of the one-carbon unit for cell metabolism. Mol Cell Biol 22: 5793–5800.
Oskarsson T, Essers MA, Dubois N, Offner S, Dubey C, Roger C et al. (2006). Skin epidermis lacking the c-Myc gene is resistant to Ras-driven tumorigenesis but can reacquire sensitivity upon additional loss of the p21Cip1 gene. Genes Dev 20: 2024–2029.
Patel JH, Loboda AP, Showe MK, Showe LC, McMahon SB . (2004). Analysis of genomic targets reveals complex functions of MYC. Nat Rev Cancer 4: 562–568.
Paz MF, Fraga MF, Avila S, Guo M, Pollan M, Herman JG et al. (2003). A systematic profile of DNA methylation in human cancer cell lines. Cancer Res 63: 1114–1121.
Pelengaris S, Littlewood T, Khan M, Elia G, Evan G . (1999). Reversible activation of c-Myc in skin: induction of a complex neoplastic phenotype by a single oncogenic lesion. Mol Cell 3: 565–577.
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.
Ponzielli R, Katz S, Barsyte-Lovejoy D, Penn LZ . (2005). Cancer therapeutics: targeting the dark side of Myc. Eur J Cancer 41: 2485–2501.
Prathapam T, Tegen S, Oskarsson T, Trumpp A, Martin GS . (2006). Activated Src abrogates the Myc requirement for the G0/G1 transition but not for the G1/S transition. Proc Natl Acad Sci USA 103: 2695–2700.
Prochownik EV, Kukowska J, Rodgers C . (1988). c-myc antisense transcripts accelerate differentiation and inhibit G1 progression in murine erythroleukemia cells. Mol Cell Biol 8: 3683–3695.
Prochownik EV . (2004). c-Myc as a therapeutic target in cancer. Expert Rev Anticancer Ther 4: 289–302.
Rothermund K, Rogulski K, Fernandes E, Whiting A, Sedivy J, Pu L et al. (2005). c-Myc-independent restoration of multiple phenotypes by two C-Myc target genes with overlapping functions. Cancer Res 65: 2097–2107.
Sharma SV, Fischbach MA, Haber DA, Settleman J . (2006). ‘Oncogenic shock’: explaining oncogene addiction through differential signal attenuation. Clin Cancer Res 12: 4392s–4395s.
Skorski T, Nieborowska-Skorska M, Campbell K, Iozzo RV, Zon G, Darzynkiewicz Z et al. (1995). Leukemia treatment in severe combined immunodeficiency mice by antisense oligodeoxynucleotides targeting cooperating oncogenes. J Exp Med 182: 1645–1653.
Soengas MS, Capodieci P, Polsky D, Mora J, Esteller M, Opitz-Araya X et al. (2001). Inactivation of the apoptosis effector Apaf-1 in malignant melanoma. Nature 409: 207–211.
Stark GR, Taylor WR . (2006). Control of the G2/M transition. Mol Biotechnol, 2006 32: 227–248.
Trumpp A, Refaeli Y, Oskarsson T, Gasser S, Murphy M, Martin GR et al. (2001). c-Myc regulates mammalian body size by controlling cell number but not cell size. Nature 414: 768–773.
Vita M, Henriksson M . (2006). The Myc oncoprotein as a therapeutic target for human cancer. Semin Cancer Biol 16: 318–330.
Yin X, Grove L, Rogulski K, Prochownik EV . (2002). Myc target in myeloid cells-1, a novel c-Myc target, recapitulates multiple c-Myc phenotypes. J Biol Chem 277: 19998–20010.
Zhou ZQ, Hurlin PJ . (2001). The interplay between Mad and Myc in proliferation and differentiation. Trends Cell Biol 11: S10–S14.
Acknowledgements
We are grateful to Dr Gary Fisher, Dr Andrzej Dlugosz and Dr Laure Rittie for critical reading of the manuscript. We thank John Lazo, David Hackham and Yatin Vyas for cell lines, Jie Lu for DNA sequencing, Josh Solomon for assistance in the initial optimization of lentiviral infection protocols and MaryBeth Ribblet and Suresh Patil for technical assistance. This work was supported by Dermatology Research Foundation Carrier Development Award to MAN, NIH grants R01-CA120244-A1 to MAN, RO1-CA105033, RO1-CA078259 to EVP and R01-CA107237 to MSS. MAN is a Melanoma Research Foundation Scholar. HW was supported by a postdoctoral fellowship award from the Research Advisory Committee of Children's Hospital of Pittsburgh.
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
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Wang, H., Mannava, S., Grachtchouk, V. et al. c-Myc depletion inhibits proliferation of human tumor cells at various stages of the cell cycle. Oncogene 27, 1905–1915 (2008). https://doi.org/10.1038/sj.onc.1210823
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DOI: https://doi.org/10.1038/sj.onc.1210823
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