Abstract
Checkpoint kinase 1 (Chk1) is a key element in the DNA-damage response pathway that is required for maintaining genomic stability. To study the potential role of Chk1 in mammary tumorigenesis, we disrupted it using a Cre/loxP system. We showed that although Chk1 heterozygosity caused abnormal development of the mammary gland, it was not sufficient to induce tumorigenesis. Simultaneous deletion of one copy of p53 failed to rescue the developmental defects; however, it synergistically induced mammary tumor formation in Chk1+/−;MMTV-Cre animals with a median time to tumor latency of about 10 months. Chk1 deficiency caused a preponderance of abnormalities, including prolongation, multipolarity, misalignment, mitotic catastrophe and loss of spindle checkpoint, that are accompanied by reduced expression of several cell cycle regulators, including Mad2. On the other hand, we also showed that Chk1 deficiency inhibited mammary tumor formation in mice carrying a homozygous deletion of p53, uncovering a complex relationship between Chk1 and p53. Furthermore, inhibition of Chk1 with a specific inhibitor, SB-218078, or acute deletion of Chk1 using small hairpin RNA killed mammary tumor cells effectively. These data show that Chk1 is critical for maintaining genome integrity and serves as a double-edged sword for cancer: although its inhibition kills cancer cells, it also triggers tumorigenesis when favorable mutations are accumulated for cell growth.
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References
Bartek J, Lukas J . (2003). Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell 3: 421–429.
Blagden S, de Bono J . (2005). Drugging cell cycle kinases in cancer therapy. Curr Drug Targets 6: 325–335.
Brodie SG, Xu X, Li C, Kuo A, Leder P, Deng CX . (2001a). Inactivation of p53 tumor suppressor gene acts synergistically with c-neu oncogene in salivary gland tumorigenesis. Oncogene 20: 1445–1454.
Brodie SG, Xu X, Qiao W, Li WM, Cao L, Deng CX . (2001b). Multiple genetic changes are associated with mammary tumorigenesis in Brca1 conditional knockout mice. Oncogene 20: 7514–7523.
Brugarolas J, Chandrasekaran C, Gordon JI, Beach D, Jacks T, Hannon GJ . (1995). Radiation-induced cell cycle arrest compromised by p21 deficiency. Nature 377: 552–557.
Carrassa L, Sanchez Y, Erba E, Damia G . (2009). U2OS cells lacking Chk1 undergo aberrant mitosis and fail to activate the spindle checkpoint. J Cell Mol Med 13: 1565–1576.
Collura A, Blaisonneau J, Baldacci G, Francesconi S . (2005). The fission yeast Crb2/Chk1 pathway coordinates the DNA damage and spindle checkpoint in response to replication stress induced by topoisomerase I inhibitor. Mol Cell Biol 25: 7889–7899.
Deng C, Zhang P, Harper JW, Elledge SJ, Leder P . (1995). Mice lacking p21CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control. Cell 82: 675–684.
Deng CX . (2006). BRCA1: cell cycle checkpoint, genetic instability, DNA damage response, and cancer evolution. Nucleic Acids Res 34: 1416–1426.
Deng CX, Wang RH . (2003). Roles of BRCA1 in DNA damage repair: a link between development and cancer. Hum Mol Genet 12: R113–R123.
Deng CX, Xu X . (2004). Generation and analysis of Brca1 conditional knockout mice. Methods Mol Biol 280: 185–200.
Dobles M, Liberal V, Scott ML, Benezra R, Sorger PK . (2000). Chromosome missegregation and apoptosis in mice lacking the mitotic checkpoint protein Mad2. Cell 101: 635–645.
Durkin SG, Arlt MF, Howlett NG, Glover TW . (2006). Depletion of CHK1, but not CHK2, induces chromosomal instability and breaks at common fragile sites. Oncogene 25: 4381–4388.
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.
Fuse E, Tanii H, Kurata N, Kobayashi H, Shimada Y, Tamura T et al. (1998). Unpredicted clinical pharmacology of UCN-01 caused by specific binding to human alpha1-acid glycoprotein. Cancer Res 58: 3248–3253.
Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ . (1993). The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 75: 805–816.
Hotte SJ, Oza A, Winquist EW, Moore M, Chen EX, Brown S et al. (2006). Phase I trial of UCN-01 in combination with topotecan in patients with advanced solid cancers: a Princess Margaret Hospital Phase II Consortium study. Ann Oncol 17: 334–340.
Jackson JR, Gilmartin A, Imburgia C, Winkler JD, Marshall LA, Roshak A . (2000). An indolocarbazole inhibitor of human checkpoint kinase (Chk1) abrogates cell cycle arrest caused by DNA damage. Cancer Res 60: 566–572.
Jonkers J, Meuwissen R, van der Gulden H, Peterse H, van der Valk M, Berns A . (2001). Synergistic tumor suppressor activity of BRCA2 and p53 in a conditional mouse model for breast cancer. Nat Genet 29: 418–425.
Kastan MB, Bartek J . (2004). Cell-cycle checkpoints and cancer. Nature 432: 316–323.
Kortmansky J, Shah MA, Kaubisch A, Weyerbacher A, Yi S, Tong W et al. (2005). Phase I trial of the cyclin-dependent kinase inhibitor and protein kinase C inhibitor 7-hydroxystaurosporine in combination with Fluorouracil in patients with advanced solid tumors. J Clin Oncol 23: 1875–1884.
Lam MH, Liu Q, Elledge SJ, Rosen JM . (2004). Chk1 is haploinsufficient for multiple functions critical to tumor suppression. Cancer Cell 6: 45–59.
Li W, Xiao C, Vonderhaar BK, Deng CX . (2007). A role of estrogen/ERalpha signaling in BRCA1-associated tissue-specific tumor formation. Oncogene 26: 7204–7212.
Liu Q, Guntuku S, Cui XS, Matsuoka S, Cortez D, Tamai K et al. (2000). Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint. Genes Dev 14: 1448–1459.
Lowe SW, Bodis S, McClatchey A, Remington L, Ruley HE, Fisher DE et al. (1994). p53 status and the efficacy of cancer therapy in vivo. Science 266: 807–810.
Margolis RL . (2005). Tetraploidy and tumor development. Cancer Cell 8: 353–354.
Michel LS, Liberal V, Chatterjee A, Kirchwegger R, Pasche B, Gerald W et al. (2001). MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells. Nature 409: 355–359.
Miki Y, Swensen J, Shattuck-Eidens D, Futreal PA, Harshman K, Tavtigian S et al. (1994). A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. Science 266: 66–71.
Muller WJ, Sinn E, Pattengale PK, Wallace R, Leder P . (1988). Single-step induction of mammary adenocarcinoma in transgenic mice bearing the activated c-neu oncogene. Cell 54: 105–115.
Padilla-Nash HM, Barenboim-Stapleton L, Difilippantonio MJ, Ried T . (2006). Spectral karyotyping analysis of human and mouse chromosomes. Nat Protoc 1: 3129–3142.
Peddibhotla S, Lam MH, Gonzalez-Rimbau M, Rosen JM . (2009). The DNA-damage effector checkpoint kinase 1 is essential for chromosome segregation and cytokinesis. Proc Natl Acad Sci USA 106: 5159–5164.
Puc J, Keniry M, Li HS, Pandita TK, Choudhury AD, Memeo L et al. (2005). Lack of PTEN sequesters CHK1 and initiates genetic instability. Cancer Cell 7: 193–204.
Puc J, Parsons R . (2005). PTEN loss inhibits CHK1 to cause double stranded-DNA breaks in cells. Cell Cycle 4: 927–929.
Shen SX, Weaver Z, Xu X, Li C, Weinstein M, Chen L et al. (1998). A targeted disruption of the murine Brca1 gene causes gamma-irradiation hypersensitivity and genetic instability. Oncogene 17: 3115–3124.
Sudakin V, Chan GK, Yen TJ . (2001). Checkpoint inhibition of the APC/C in HeLa cells is mediated by a complex of BUBR1, BUB3, CDC20, and MAD2. J Cell Biol 154: 925–936.
Takai H, Tominaga K, Motoyama N, Minamishima YA, Nagahama H, Tsukiyama T et al. (2000). Aberrant cell cycle checkpoint function and early embryonic death in Chk1(−/−) mice. Genes Dev 14: 1439–1447.
Tang Z, Bharadwaj R, Li B, Yu H . (2001). Mad2-Independent inhibition of APCCdc20 by the mitotic checkpoint protein BubR1. Dev Cell 1: 227–237.
Wagner KU, Wall RJ, St-Onge L, Gruss P, Wynshaw-Boris A, Garrett L et al. (1997). Cre-mediated gene deletion in the mammary gland. Nucleic Acids Res 25: 4323–4330.
Wang RH, Yu H, Deng CX . (2004). A requirement for breast-cancer-associated gene 1 (BRCA1) in the spindle checkpoint. Proc Natl Acad Sci USA 101: 17108–17113.
Wang X, Zhou YX, Qiao W, Tominaga Y, Ouchi M, Ouchi T et al. (2006). Overexpression of aurora kinase A in mouse mammary epithelium induces genetic instability preceding mammary tumor formation. Oncogene 25: 7148–7158.
Xiao Z, Chen Z, Gunasekera AH, Sowin TJ, Rosenberg SH, Fesik S et al. (2003). Chk1 mediates S and G2 arrests through Cdc25A degradation in response to DNA-damaging agents. J Biol Chem 278: 21767–21773.
Xu X, Qiao W, Linke SP, Cao L, Li WM, Furth PA et al. (2001). Genetic interactions between tumor suppressors Brca1 and p53 in apoptosis, cell cycle and tumorigenesis. Nat Genet 28: 266–271.
Xu X, Weaver Z, Linke SP, Li C, Gotay J, Wang XW et al. (1999). Centrosome amplification and a defective G2-M cell cycle checkpoint induce genetic instability in BRCA1 exon 11 isoform-deficient cells. Mol Cell 3: 389–395.
Yarden RI, Brody LC . (2001). Identification of proteins that interact with BRCA1 by Far-Western library screening. J Cell Biochem 83: 521–531.
Yarden RI, Pardo-Reoyo S, Sgagias M, Cowan KH, Brody LC . (2002). BRCA1 regulates the G2/M checkpoint by activating Chk1 kinase upon DNA damage. Nat Genet 30: 285–289.
Zachos G, Black EJ, Walker M, Scott MT, Vagnarelli P, Earnshaw WC et al. (2007). Chk1 is required for spindle checkpoint function. Dev Cell 12: 247–260.
Zhang WH, Poh A, Fanous AA, Eastman A . (2008). DNA damage-induced S phase arrest in human breast cancer depends on Chk1, but G2 arrest can occur independently of Chk1, Chk2 or MAPKAPK2. Cell Cycle 7: 1668–1677.
Zhao H, Watkins JL, Piwnica-Worms H . (2002). Disruption of the checkpoint kinase 1/cell division cycle 25A pathway abrogates ionizing radiation-induced S and G2 checkpoints. Proc Natl Acad Sci USA 99: 14795–14800.
Acknowledgements
We gratefully acknowledge members of Deng laboratory for a critical reading of the article. This work was supported by the Intramural Research Program of the National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, USA.
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Fishler, T., Li, YY., Wang, RH. et al. Genetic instability and mammary tumor formation in mice carrying mammary-specific disruption of Chk1 and p53. Oncogene 29, 4007–4017 (2010). https://doi.org/10.1038/onc.2010.163
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DOI: https://doi.org/10.1038/onc.2010.163
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