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A small molecule based on the pRb2/p130 spacer domain leads to inhibition of cdk2 activity, cell cycle arrest and tumor growth reduction in vivo

Oncogene volume 26, pages 1829–1839 (2007)Cite this article

Abstract

One strategy in the development of anticancer therapeutics has been to arrest malignant proliferation through inhibition of the enzymatic activity of cyclin-dependent kinases (cdks), which are key regulatory molecules of the cell cycle. Over the past few years, numerous compounds with remarkable cdk inhibitory activity have been studied in cancer therapy, although it is very difficult to point out the best cdk to target. An excellent candidate appears to be cdk2, whose alteration is a pathogenic hallmark of tumorigenesis. The small molecule described in our study showed an inhibitory effect on the kinase activity of cdk2, a significant growth arrest observed in a colony formation assay and a reduction in the size of the tumor in nude mice, thus suggesting its potential role as a promising new type of mechanism-based antitumor drug, also for the treatment of hyperproliferative disorders.

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References

  • Al-Aynati MM, Radulovich N, Ho J, Tsao MS . (2004). Overexpression of G1–S cyclins and cyclin-dependent kinases during multistage human pancreatic duct cell carcinogenesis. Clin Cancer Res 10: 6598–6605.

    Article  CAS  PubMed  Google Scholar 

  • Alessi F, Quarta S, Savio M, Riva F, Rossi L, Stivala LA et al. (1998). The cyclin-dependent kinase inhibitors olomoucine and roscovitine arrest human fibroblasts in G1 phase by specific inhibition of CDK2 kinase activity. Exp Cell Res 245: 8–18.

    Article  CAS  PubMed  Google Scholar 

  • Andrews MJ, McInnes C, Kontopidis G, Innes L, Cowan A, Plater A et al. (2004). Design, synthesis, biological activity and structural analysis of cyclic peptide inhibitors targeting the substrate recruitment site of cyclin-dependent kinase complexes. Org Biomol Chem 2: 2735–2741.

    Article  CAS  PubMed  Google Scholar 

  • Bagella L, Giacinti C, Simone C, Giordano A . (2006). Identification of murine cdk10: association with Ets2 transcription factor and effects on the cell cycle. J Cell Biochem (E-pub ahead of print).

  • Bates S, Bonetta L, MacAllan D, Parry D, Holder A, Dickson C et al. (1994). CDK6 (PLSTIRE) and CDK4 (PSK-J3) are a distinct subset of the cyclin-dependent kinases that associate with cyclin D1. Oncogene 9: 71–79.

    CAS  PubMed  Google Scholar 

  • Berthet C, Aleem E, Coppola V, Tessarollo L, Kaldis P . (2003). Cdk2 knockout mice are viable. Curr Biol 13: 1775–1785.

    Article  CAS  PubMed  Google Scholar 

  • Caputi M, Russo G, Esposito V, Mancini A, Giordano A . (2005). Role of cell-cycle regulators in lung cancer. J Cell Physiol 205: 319–327.

    Article  CAS  PubMed  Google Scholar 

  • Carlson BA, Dubay MM, Sausville EA, Brizuela L, Worland PJ . (1996). Flavopiridol induces G1 arrest with inhibition of cyclin-dependent kinase (CDK) 2 and CDK4 in human breast carcinoma cells. Cancer Res 56: 2973–2978.

    CAS  PubMed  Google Scholar 

  • Chae HD, Yun J, Bang YJ, Shin DY . (2004). Cdk2-dependent phosphorylation of the NF-Y transcription factor is essential for the expression of the cell cycle-regulatory genes and cell cycle G1/S and G2/M transitions. Oncogene 23: 4084–4088.

    Article  CAS  PubMed  Google Scholar 

  • Chao SH, Fujinaga K, Marion JE, Taube R, Sausville EA, Senderowicz AM et al. (2000). Flavopiridol inhibits P-TEFb and blocks HIV-1 replication. J Biol Chem 275: 28345–28348.

    Article  CAS  PubMed  Google Scholar 

  • Cinti C, Macaluso M, Giordano A . (2005). Tumor-specific exon 1 mutations could be the ‘hit event’ predisposing Rb2/p130 gene to epigenetic silencing in lung cancer. Oncogene 24: 5821–5826.

    Article  PubMed  Google Scholar 

  • Claudio PP, Howard CM, Baldi A, De Luca A, Fu Y, Condorelli G et al. (1994). p130/pRb2 has growth suppressive properties similar to yet distinctive from those of retinoblastoma family members pRb and p107. Cancer Res 54: 5556–5560.

    CAS  PubMed  Google Scholar 

  • Claudio PP, De Luca A, Howard CM, Baldi A, Firpo EJ, Koff A et al. (1996). Functional analysis of pRb2/p130 interaction with cyclins. Cancer Res 56: 2003–2008.

    CAS  PubMed  Google Scholar 

  • Claudio PP, Howard CM, Pacilio C, Cinti C, Romano G, Minimo C et al. (2000). Mutations in the retinoblastoma-related gene RB2/p130 in lung tumors and suppression of tumor growth in vivo by retrovirus-mediated gene transfer. Cancer Res 60: 372–382.

    CAS  PubMed  Google Scholar 

  • Dai Y, Grant S . (2004). Small molecule inhibitors targeting cyclin-dependent kinases as anticancer agents. Curr Oncol Rep 6: 123–130.

    Article  PubMed  Google Scholar 

  • De Luca A, MacLachlan TK, Bagella L, Dean C, Howard CM, Claudio PP et al. (1997). A unique domain of pRb2/p130 acts as an inhibitor of Cdk2 kinase activity. J Biol Chem 272: 20971–20974.

    Article  CAS  PubMed  Google Scholar 

  • Du W, Pogoriler J . (2006). Retinoblastoma family genes. Oncogene 25: 5190–5200.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fischer PM, Gianella-Borradori A . (2003). CDK inhibitors in clinical development for the treatment of cancer. Expert Opin Investig Drugs 12: 955–970.

    Article  CAS  PubMed  Google Scholar 

  • Gallo G, Giordano A . (2005). Are RB proteins a potential substrate of Pin1 in the regulation of the cell cycle? J Cell Physiol 205: 176–181.

    Article  CAS  PubMed  Google Scholar 

  • Geng Y, Yu Q, Sicinska E, Das M, Schneider JE, Bhattacharya S et al. (2003). Cyclin E ablation in the mouse. Cell 114: 431–443.

    Article  CAS  PubMed  Google Scholar 

  • Genovese C, Trani D, Caputi M, Claudio PP . (2006). Cell cycle control and beyond: emerging roles for the retinoblastoma gene family. Oncogene 25: 5201–5209.

    Article  CAS  PubMed  Google Scholar 

  • Gibbs JB, Oliff A . (1994). Pharmaceutical research in molecular oncology. Cell 79: 193–198.

    Article  CAS  PubMed  Google Scholar 

  • Giordano A, Lee JH, Scheppler JA, Herrmann C, Harlow E, Deuschle U et al. (1991). Cell cycle regulation of histone H1 kinase activity associated with the adenoviral protein E1A. Science 253: 1271–1275.

    Article  CAS  PubMed  Google Scholar 

  • Giordano A, Whyte P, Harlow E, Franza Jr BR, Beach D, Draetta G . (1989). A 60 kd cdc2-associated polypeptide complexes with the E1A proteins in adenovirus-infected cells. Cell 58: 981–990.

    Article  CAS  PubMed  Google Scholar 

  • Gray NS, Wodicka L, Thunnissen AM, Norman TC, Kwon S, Espinoza FH et al. (1998). Exploiting chemical libraries, structure, and genomics in the search for kinase inhibitors. Science 281: 533–538.

    Article  CAS  PubMed  Google Scholar 

  • Gu Y, Rosenblatt J, Morgan DO . (1992). Cell cycle regulation of CDK2 activity by phosphorylation of Thr160 and Tyr15. EMBOJ 11: 3995–4005.

    Article  CAS  Google Scholar 

  • Harwell RM, Mull BB, Porter DC, Keyomarsi K . (2004). Activation of cyclin-dependent kinase 2 by full length and low molecular weight forms of cyclin E in breast cancer cells. J Biol Chem 279: 12695–12705.

    Article  CAS  PubMed  Google Scholar 

  • Howard CM, Claudio PP, De Luca A, Stiegler P, Jori FP, Safdar NM et al. (2000). Inducible pRb2/p130 expression and growth-suppressive mechanisms: evidence of a pRb2/p130, p27Kip1, and cyclin E negative feedback regulatory loop. Cancer Res 60: 2737–2744.

    CAS  PubMed  Google Scholar 

  • Hsu YL, Kuo PL, Lin CC . (2004). Proliferative inhibition, cell-cycle dysregulation, and induction of apoptosis by ursolic acid in human non-small cell lung cancer A549 cells. Life Sci 75: 2303–2316.

    Article  CAS  PubMed  Google Scholar 

  • Kamb A, Gruis NA, Weaver-Feldhaus J, Liu Q, Harshman K, Tavtigian SV et al. (1994). A cell cycle regulator potentially involved in genesis of many tumor types. Science 264: 436–440.

    Article  CAS  PubMed  Google Scholar 

  • Koff A, Giordano A, Desai D, Yamashita K, Harper JW, Elledge S et al. (1992). Formation and activation of a cyclin E-cdk2 complex during the G1 phase of the human cell cycle. Science 257: 1689–1694.

    Article  CAS  PubMed  Google Scholar 

  • Lee HR, Chang TH, Tebalt III MJ, Senderowicz AM, Szabo E . (1999). Induction of differentiation accompanies inhibition of Cdk2 in a non-small cell lung cancer cell line. Int J Oncol 15: 161–166.

    CAS  PubMed  Google Scholar 

  • Lees JA, Buchkovich KJ, Marshak DR, Anderson CW, Harlow E . (1991). The retinoblastoma protein is phosphorylated on multiple sites by human cdc2. EMBOJ 10: 4279–4290.

    Article  CAS  Google Scholar 

  • Li Y, Chinni SR, Senderowicz AM, Sarkar FH . (2000). Induction of growth inhibition and apoptosis in prostate cancer cells by flavopiridol. Int J Oncol 17: 755–759.

    CAS  PubMed  Google Scholar 

  • Liao C, Li S, Wang X, Muhlrad S, Bjartell A, Wolgemuth DJ . (2004). Elevated levels and distinct patterns of expression of A-type cyclins and their associated cyclin-dependent kinases in male germ cell tumors. Int J Cancer 108: 654–664.

    Article  CAS  PubMed  Google Scholar 

  • Losiewicz MD, Carlson BA, Kaur G, Sausville EA, Worland PJ . (1994). Potent inhibition of CDC2 kinase activity by the flavonoid L86-8275. Biochem Biophys Res Commun 201: 589–595.

    Article  CAS  PubMed  Google Scholar 

  • Macaluso M, Montanari M, Giordano A . (2006). Rb family proteins as modulators of gene expression and new aspects regarding the interaction with chromatin remodeling enzymes. Oncogene 25: 5263–5267.

    Article  CAS  PubMed  Google Scholar 

  • MacLachlan TK, Sang N, Giordano A . (1995). Cyclins, cyclin-dependent kinases and cdk inhibitors: implications in cell cycle control and cancer. Crit Rev Eukaryot Gene Expr 5: 127–156.

    Article  CAS  PubMed  Google Scholar 

  • Malumbres M, Barbacid M . (2005). Mammalian cyclin-dependent kinases. Trends Biochem Sci 30: 630–641.

    Article  CAS  PubMed  Google Scholar 

  • Matsumoto Y, Hayashi K, Nishida E . (1999). Cyclin-dependent kinase 2 (Cdk2) is required for centrosome duplication in mammalian cells. Curr Biol 9: 429–432.

    Article  CAS  PubMed  Google Scholar 

  • Matsumoto Y, Maller JL . (2004). A centrosomal localization signal in cyclin E required for Cdk2-independent S phase entry. Science 306: 885–888.

    Article  CAS  PubMed  Google Scholar 

  • Mayol X, Grana X, Baldi A, Sang N, Hu Q, Giordano A . (1993). Cloning of a new member of the retinoblastoma gene family (pRb2) which binds to the E1A transforming domain. Oncogene 8: 2561–2566.

    CAS  PubMed  Google Scholar 

  • Merola E, Mattioli E, Minimo C, Zuo W, Rabitti C, Cicala M et al. (2006). Immunohistochemical evaluation of pRb2/p130, VEGF, EZH2, p53, p16, p21waf-1, p27, and PCNA in Barrett's esophagus. J Cell Physiol 207: 512–519.

    Article  CAS  PubMed  Google Scholar 

  • Morgan DO . (1997). Cyclin-dependent kinases: engines, clocks, and microprocessors. Annu Rev Cell Dev Biol 13: 261–291.

    Article  CAS  PubMed  Google Scholar 

  • Nobori T, Miura K, Wu DJ, Lois A, Takabayashi K, Carson DA . (1994). Deletions of the cyclin-dependent kinase-4 inhibitor gene in multiple human cancers. Nature 368: 753–756.

    Article  CAS  PubMed  Google Scholar 

  • Ohtsubo M, Theodoras AM, Schumacher J, Roberts J, Pagano M . (1995). Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol Cell Biol 15: 2612–2624.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Olofsson A, Willen H, Goransson M, Engstrom K, Meis-Kindblom JM, Stenman G et al. (2004). Abnormal expression of cell cycle regulators in FUS-CHOP carrying liposarcomas. Int J Oncol 25: 1349–1355.

    CAS  PubMed  Google Scholar 

  • Ortega S, Prieto I, Odajima J, Martin A, Dubus P, Sotillo R et al. (2003). Cyclin-dependent kinase 2 is essential for meiosis but not for mitotic cell division in mice. Nat Genet 35: 25–31.

    Article  CAS  PubMed  Google Scholar 

  • Pepper C, Thomas A, Hoy T, Fegan C, Bentley P . (2001). Flavopiridol circumvents Bcl-2 family mediated inhibition of apoptosis and drug resistance in B-cell chronic lymphocytic leukaemia. Br J Haematol 114: 70–77.

    Article  CAS  PubMed  Google Scholar 

  • Purev E, Giordano A, Soprano DR, Soprano KJ . (2006). Interaction of PP2A catalytic subunit with Rb2/p130 is required for all-trans retinoic acid suppression of ovarian carcinoma cell growth. J Cell Physiol 206: 495–502.

    Article  CAS  PubMed  Google Scholar 

  • Resnitzky D, Reed SI . (1995). Different roles for cyclins D1 and E in regulation of the G1-to-S transition. Mol Cell Biol 15: 3463–3469.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Senderowicz AM . (2003). Small-molecule cyclin-dependent kinase modulators. Oncogene 22: 6609–6620.

    Article  CAS  PubMed  Google Scholar 

  • Shapiro GI . (2006). Cyclin-dependent kinase pathways as targets for cancer treatment. J Clin Oncol 24: 1770–1783.

    Article  CAS  PubMed  Google Scholar 

  • Song Y, Kesuma D, Wang J, Deng Y, Duan J, Wang JH et al. (2004). Specific inhibition of cyclin-dependent kinases and cell proliferation by harmine. Biochem Biophys Res Commun 317: 128–132.

    Article  CAS  PubMed  Google Scholar 

  • Sunters A, Thomas DP, Yeudall WA, Grigoriadis AE . (2004). Accelerated cell cycle progression in osteoblasts overexpressing the c-fos proto-oncogene: induction of cyclin A and enhanced CDK2 activity. J Biol Chem 279: 9882–9891.

    Article  PubMed  Google Scholar 

  • Tetsu O, McCormick F . (2003). Proliferation of cancer cells despite CDK2 inhibition. Cancer Cell 3: 233–245.

    Article  CAS  PubMed  Google Scholar 

  • Tosi GM, Trimarchi C, Macaluso M, la Sala D, Ciccodicola A, Lazzi S et al. (2005). Genetic and epigenetic alterations of RB2/p130 tumor suppressor gene in human sporadic retinoblastoma: implications for pathogenesis and therapeutic approach. Oncogene 24: 5827–5836.

    Article  CAS  PubMed  Google Scholar 

  • Vives E, Brodin P, Lebleu B . (1997). A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus. J Biol Chem 272: 16010–16017.

    Article  CAS  PubMed  Google Scholar 

  • Weinberg RA . (1995). The retinoblastoma protein and cell cycle control. Cell 81: 323–330.

    Article  CAS  PubMed  Google Scholar 

  • Woo MS, Sanchez I, Dynlacht BD . (1997). p130 and p107 use a conserved domain to inhibit cellular cyclin-dependent kinase activity. Mol Cell Biol 17: 3566–3579.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xiong Y . (1996). Why are there so many CDK inhibitors? Biochim Biophys Acta 1288: 01–05.

    CAS  PubMed  Google Scholar 

  • Yu CH, Wu J, Su YF, Ho PY, Liang YC, Sheu MT et al. (2004). Anti-proliferation effect of 3-amino-2-imino-3,4-dihydro-2H-1,3-benzothiazin-4-one (BJ-601) on human vascular endothelial cells: G0/G1 p21-associated cell cycle arrest. Biochem Pharmacol 67: 1907–1916.

    Article  CAS  PubMed  Google Scholar 

  • Zhu L, Enders G, Lees JA, Beijersbergen RL, Bernards R, Harlow E . (1995a). The pRB-related protein p107 contains two growth suppression domains: independent interactions with E2F and cyclin/cdk complexes. EMBOJ 14: 1904–1913.

    Article  CAS  Google Scholar 

  • Zhu L, Harlow E, Dynlacht BD . (1995b). p107 uses a p21CIP1-related domain to bind cyclin/cdk2 and regulate interactions with E2F. Genes Dev 9: 1740–1752.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to Ms M Basso for her assistance in editing the manuscript. This work was supported by Sbarro Health Research Organization and NIH grants (AG).

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Authors and Affiliations

  1. Sbarro Institute for Cancer Research and Molecular Medicine, Centre of Biotechnology, Temple University, Philadelphia, PA, USA

    L Bagella, A Sun, T Tonini, G Abbadessa, A De Luca, P P Claudio & A Giordano

  2. Department of Biomedical Sciences, Division of Biochemistry, University of Sassari, Sassari, Italy

    L Bagella

  3. Department of Human Pathology and Oncology, University of Siena, Siena, Italy

    T Tonini & A Giordano

  4. Department for the Development of Therapeutic Programs, Laboratory ‘C’, Center for Experimental Research, Regina Elena Cancer Institute, Rome, Italy

    G Cottone & M G Paggi

  5. Department of Medicine and Public Health, Section of Clinical Anatomy, Second University of Naples, Naples, Italy

    A De Luca

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  1. L Bagella
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Correspondence to A Giordano.

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Bagella, L., Sun, A., Tonini, T. et al. A small molecule based on the pRb2/p130 spacer domain leads to inhibition of cdk2 activity, cell cycle arrest and tumor growth reduction in vivo. Oncogene 26, 1829–1839 (2007). https://doi.org/10.1038/sj.onc.1209987

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