Cancer Cell Biology
Transient arrest in a quiescent state allows ovarian cancer cells to survive suboptimal growth conditions and is mediated by both Mirk/dyrk1b and p130/Rb2
Jing Hu,
Hassan Nakhla,
Eileen Friedman,
Jing Hu
Pathology Department, Upstate Medical University, State University of New York, Syracuse, New York
Search for more papers by this authorHassan Nakhla
Pathology Department, Upstate Medical University, State University of New York, Syracuse, New York
Search for more papers by this authorCorresponding Author
Eileen Friedman
Pathology Department, Upstate Medical University, State University of New York, Syracuse, New York
Pathology Department, Upstate Medical University, Syracuse, NY 13210, USA, Tel.: 315-464-7148Search for more papers by this authorJing Hu,
Hassan Nakhla,
Eileen Friedman,
Jing Hu
Pathology Department, Upstate Medical University, State University of New York, Syracuse, New York
Search for more papers by this authorHassan Nakhla
Pathology Department, Upstate Medical University, State University of New York, Syracuse, New York
Search for more papers by this authorCorresponding Author
Eileen Friedman
Pathology Department, Upstate Medical University, State University of New York, Syracuse, New York
Pathology Department, Upstate Medical University, Syracuse, NY 13210, USA, Tel.: 315-464-7148Search for more papers by this authorAbstract
Some ovarian cancer cells in vivo are in a reversible quiescent state where they can contribute to cancer spread under favorable growth conditions. The serine/threonine kinase Mirk/dyrk1B was expressed in each of seven ovarian cancer cell lines and in 21 of 28 resected human ovarian cancers, and upregulated in 60% of the cancers. Some ovarian cancer cells were found in a G0 quiescent state, with the highest fraction in a line with an amplified Mirk gene. Suboptimal culture conditions increased the G0 fraction in SKOV3 and TOV21G, but not OVCAR4 cultures. Less than half as many OVCAR4 cells survived under suboptimal culture conditions as shown by total cell numbers, dye exclusion viability studies, and assay of cleaved apoptotic marker proteins. G0 arrest in TOV21G and SKOV3 cells led to increased levels of Mirk, the CDK inhibitor p27, p130/Rb2, and p130/Rb2 complexed with E2F4. The G0 arrest was transient, and cells exited G0 when fresh nutrients were supplied. Depletion of p130/Rb2 reduced the G0 fraction, increased cell sensitivity to serum-free culture and to cisplatin, and reduced Mirk levels. Mirk contributed to G0 arrest by destabilization of cyclin D1. In TOV21G cells, but not in normal diploid fibroblasts, Mirk depletion led to increased apoptosis and loss of viability. Because Mirk is expressed at low levels in most normal adult tissues, the elevated Mirk protein levels in ovarian cancers may present a novel therapeutic target, in particular for quiescent tumor cells which are difficult to eradicate by conventional therapies targeting dividing cells.
Supporting Information
Additional Supporting Information may be found in the online version of this article.
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| IJC_25692_sm_suppfigure1.tif95.8 KB | Supporting Figure 1. |
| IJC_25692_sm_suppfigure2.tif138.2 KB | Supporting Figure 2. |
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References
- 1 Deng X, Mercer SE, Shah S, Ewton DZ, Friedman E. The Cyclin-dependent Kinase Inhibitor p27Kip1 Is Stabilized in G0 by Mirk/dyrk1B Kinase. J Biol Chem 2004; 279: 22498–504.
- 2 Besson A, Gurian-West M, Chen X, Kelly-Spratt KS, Kemp CJ, Roberts JM. A pathway in quiescent cells that controls p27Kip1 stability, subcellular localization, and tumor suppression. Genes Dev 2006; 20: 47–64.
- 3 Janumyan Y, Cui Q, Yan L, Sansam CG, Vanlentin M, Yang E. G0 function of BCL2 and BCL-xL requires BAX, BAK, and p27 phosphorylation by Mirk, revealing a novel role of BAX and BAK in quiescence regulation. J Biol Chem 2008; 283: 34108–20.
- 4 Sang L, Coller HA, Roberts JM. Control of the reversibility of cellular quiescence by the transcriptional repressor HES1. Science 2008; 321: 1095–100.
- 5 Sang L, Roberts JM, Coller HA. Hijacking HES1: how tumors co-opt the anti-differentiation strategies of quiescent cells. Trends Mol Med 2010; 16: 17.
- 6 Barnes D, Melo J. Primitive, quiescent and difficult to kill: the role of non-proliferating stem cells in chronic myeloid leukemia. Cell Cycle 2006; 5: 2862–6.
- 7 Deng X, Ewton DZ, Friedman E. Mirk/Dyrk1B maintains the viability of quiescent pancreatic cancer cells by decreasing ROS levels. Cancer Res 2009; 69: 3317–24.
- 8 Schindlbeck C, Hantschmann P, Zerzer M, Jahns B, Rjosk D, Janni W, Rack, B Sommer H, Friese K. Prognostic impact of KI67, p53, human epithelial growth factor receptor 2, topoisomerase IIalpha, epidermal growth factor receptor, and nm23 expression of ovarian carcinomas and disseminated tumor cells in the bone marrow. Int J Gynecol Cancer 2007; 17: 1047–55.
- 9 Kusumbe AP, Bapat SA. Cancer stem cells and aneuploid populations within developing tumors are the major determinants of tumor dormancy. Cancer Res 2009; 69: 9245–53.
- 10 Lu Z, Luo R, Lu Y, Zhang X, Yu Q, Khare S, Kondo S, Kondo Y, Yu, Y. Mills GB, Liao S-L, Bast RC, Jr The tumor suppressor gene ARHI regulates autophagy and tumor dormancy in human ovarian cancer cells. J Clin Invest 2008; 118: 3917–29.
- 11 Amaravadi RK. Autophagy-induced tumor dormancy in ovarian cancer. J Clin Invest 2008; 118: 3837–40.
- 12 Adam AP, George A, Schewe D, Bragado P, Iglesias BV, Ranganathan AC, Kourtidis A, Conklin DS, Aguirre-Ghiso JA. Computational identification of a p38SAPK-regulated transcription factor network required for tumor cell quiescence. Cancer Res 2009; 69: 5664–72.
- 13 Chen C, Liu Y, Liu R, Ikenoue T, Guan K-L, Liu Y, Zheng P. TSC-mTOR maintains quiescence and function of hematopoietic stem cells by repressing mitochondrial biogenesis and reactive oxygen species. J Exp Med 2008; 205: 2397–408.
- 14 Jin K, Park S-J, Ewton D, Friedman E. The survival kinase Mirk/dyrk1B is a downstream effector of oncogenic K-ras. Cancer Res 2007; 67: 7247–55.
- 15 Tejedor F, Zhu X, Kaltenbach E, Ackermann A, Baumann A, Canal I, Heisenberg M, Fischbach KF, Pongs O. Minibrain:a new protein kinase family involved in postembryonic neurogenesis in Drosophila. Neuron 1995; 14: 287–301.
- 16 Kentrup H, Becker W, Heukelbach J, Wilmes A, Schurman A, Huppertz C, Kainulainen H, Joost H-G. Dyrk, a dual specificity protein kinase with unique structural features whose activity is dependent on tyrosine residues between subdomains VII and VIII. J Biol Chem 1996; 271: 3488–95.
- 17 Becker W, Weber Y, Wetzel K, Eirmbter K, Tejedor F, Joost H-G. Sequence characteristics, subcellular localization and substrate specificity of dyrk-related kinases, a novel family of dual specificity protein kinases. J Biol Chem 1998; 273: 25893–902.
- 18 Mercer SE, Ewton DZ, Deng X, Lim S, Mazur TR, Friedman E. Mirk/Dyrk1B Mediates Survival during the Differentiation of C2C12 Myoblasts. J Biol Chem 2005; 280: 25788–801.
- 19 Arron JR, Winslow MM, Polleri A, Chang C-P, Wu H, Gao X, Neilson JR, Chen L, Heit JJ, Kim SK, Yamasaki N, Miyakawa T, Francke U, Graef IA, Crabtree GR. NFAT dysregulation by increased dosage of DSCR1 and DYRK1A on chromosome 21. Nature 2006; 441: 595–600.
- 20 Geiger JN, Knudsen GT, Panek L, Pandit AK, Yoder MD, Lord KA, Creasy CL, Burns BM, Gaines P, Dillon SB, Wojchowski DM. mDYRK3 kinase is expressed selectively in late erythroid progenitor cells and attenuates colony-forming unit-erythroid development. Blood 2001; 97: 901–10.
- 21 Li K, Zhao S, Karur V, Wojchowski DM. DYRK3 activation, engagement of protein kinase A/cAMP response element-binding protein, and modulation of progenitor cell survival. J Biol Chem 2002; 277: 47052–60.
- 22 Sacher F, Moller C, Bone W, Gottwald U, Fritsch M. The expression of the testis-specific Dyrk4 kinase is highly restricted to step 8 spermatids but is not required for male fertility in mice. Mol Cell Endocrinol 2007; 267: 80–8.
- 23 Gwack Y, Sharma S, Nardone J, Tanasa B, Iuga A, Srikanth S, Okamura H, Bolton D, Feske S, Hogan PG, Rao A. A genome-wide Drosophila RNAi screen identifies DYRK-family kinases as regulators of NFAT. Nature 2006; 441: 646.
- 24 Leder S, Czajkowska H, Maenz B, de Graaf K, Barthel A, Joost H-G, Becker W. Alternative splicing variants of the protein kinase DYRK1B exhibit distinct patterns of expression and functional properties. Biochem J 2003; 372: 881–8.
- 25 Collins CS, Hong J, Sapinoso L, Zhou Y, Liu Z, Micklash K, Schultz, PG, Hampton G. A small interfering RNA screen for modulators of tumor cell motility identifies MAP4K4 as a promigratory kinase. PNAS 2006; 103: 3775–80.
- 26 Lee K, Deng X, Friedman E. Mirk protein kinase is a mitogen-activated protein kinase substrate that mediates survival of colon cancer cells. Cancer Res 2000; 60: 3631–37.
- 27 Mercer SE, Ewton DZ, Shah S, Naqvi A, Friedman E. Mirk/Dyrk1b mediates cell survival in rhabdomyosarcomas. Cancer Res 2006; 66: 5143–50.
- 28 Deng X, Ewton DZ, Li S, Naqvi A, Mercer SE, Landas S, Friedman E. The kinase Mirk/Dyrk1B mediates cell survival in pancreatic ductal adenocarcinoma. Cancer Res 2006; 66: 4149–58.
- 29 Deng X, Ewton DZ, Mercer SE, Friedman E. Mirk/dyrk1B decreases the nuclear accumulation of class II histone deacetylases during skeletal muscle differentiation. J Biol Chem 2005; 280: 4894–905.
- 30 Coller H, Sang L, Roberts JM. A new description of cellular quiescence. PLOS Biol 2006; 4: 329–49.
- 31 D'Andrilli G, Masciullo V, Bagella L, Tonini T, Minimo C, Zannoni GF, Giuntoli RL, II, Carlson JA, Jr, Soprano DR, Soprano KJ, Scambia G, Giordano A. Frequent loss of pRb2/p130 in human ovarian carcinoma. Clin Cancer Res 2004; 10: 3098–103.
- 32 Smith E, Leone G, Nevins J. Distinct mechanisms control the accumulation of the Rb-related p107 and p130 proteins during cell growth. Cell Growth Differ 1998; 9: 297–303.
- 33 Kops GJPL, Dansen TB, Polderman PE, Saarloos I, Wirtz KWA, Coffer PJ, Lam EW, Burgering F, Boudewijn MT. Forkhead transcription factor FOXO3a protects quiescent cells from oxidative stress. Nature 2002; 419: 316.
- 34 Canhoto A, Chestukhin A, Litovchick L, DeCaprio JA. Phosphorylation of the retinoblastoma-related protein p130 in growth-arrested cells. Oncogene 2000; 19: 5116–22.
- 35 Farkas T, Hansen K, Holm K, Lukas J, Bartek J. Distinct phosphorylation events regulate p130- and p107-mediated repression of E2F-4. J Biol Chem 2002; 277: 26741–52.
- 36 Smith EJ, Leone G, DeGregori J, Jakoi L, Nevins JR. The accumulation of an E2F-p130 transcriptional repressor distinguishes a G0 cell state from a G1 cell state. Mol Cell Biol 1996; 16: 6965–76.
- 37 Deng X, Ewton DZ, Pawlikowski B, Maimone M, Friedman E. Mirk/dyrk1B is a Rho-induced kinase active in skeletal muscle differentiation. J Biol Chem 2003; 278: 41347–54.
- 38 Jin K, Ewton D, Park S, Hu J, Friedman E. Mirk regulates the exit of colon cancer cells from quiescence. J Biol Chem 2009; 284: 22916–25.
- 39 Zou Y, Ewton D, Deng D, Mercer S, Friedman E. Mirk/dyrk1B kinase destabilizes cyclin D1 by phosphorylation at threonine 288. J Biol Chem 2004; 279: 27790–98.
- 40 Thompson FH, Nelson MA, Trent JM, Guan X-Y, Liu Y, Yang J-M, Emerson J, Adair L, Wymer J, Balfour C, Massey K, Weinstein R, Alberts DS, Taetle R. Amplification of 19q13.1-q13.2 sequences in ovarian cancer: G-band, FISH, and molecular studies. Cancer Genet Cytogenet 1996; 87: 55.
- 41 Karhu R, Mahlamaki E, Kallioniemi A. Pancreatic adenocarcinoma-genetic portrait from chromosomes to microarrays. Genes Chromosomes Cancer 2006; 45: 721–30.
- 42 Moniaux N, Nemos C, Schmied BM, Chauhan SC, Deb S, Morikane K, Choudhury A, VanLith M, Sutherlin M, Sikela JM, Hollingsworth MA, Batra SK. The human homologue of the RNA polymerase II-associated factor 1 (hPaf1), localized on the 19q13 amplicon, is associated with tumorigenesis. Oncogene 2006; 25: 3247–57.
- 43 Kuuselo R, Savinainen K, Azorsa DO, Basu GD, Karhu R, Tuzmen S, Mousses S, Kallioniemi A. Intersex-like (IXL) is a cell survival regulator in pancreatic cancer with 19q13 amplification. Cancer Res 2007; 67: 1943–49.
- 44 Cheng J, Godwin A, Bellacosa A, Taguchi T, Franke T, Hamilton T, Tsichlis PN, Testa JR. AKT2, a putative oncogene encoding a member of a subfamily of protein-serine/threonine kinases, is amplified in human ovarian carcinomas. PNAS 1992; 89: 9267–71.
- 45 Hu J, Friedman E. Mirk kinase is a novel factor mediating cisplatin resistance in ovarian cancer cells. Genes Cancer, in press.
- 46 Moreno CS, Matyunina L, Dickerson EB, Schubert N, Bowen NJ, Logani S, Benigno BB, McDonald JF. Evidence that p53 mediated cell-cycle-arrest inhibits chemotherapeutic treatment of ovarian carcinomas. PLoS ONE 2007; 2: e441.
- 47 Shaw RJ. LKB1 and AMP-activated protein kinase control of mTOR signalling and growth. Acta Physiol 2009; 196: 65–80.
- 48 Soprano KJ, Purev E, Vuocolo S, Soprano DR. Rb2/p130 and protein phosphatase 2A: key mediators of ovarian carcinoma cell growth suppression by all-trans retinoic acid. Oncogene 2006; 25: 5315.
- 49 Litovchick L, Sadasivam S, Florens L, Zhu X, Swanson SK, Velmurugan S, Chestukhin A, DeCaprio JA. Evolutionarily conserved multisubunit RBL2/p130 and E2F4 protein complex represses human cell cycle-dependent genes in quiescence. Mol Cell 2007; 26: 539.
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