Abstract
Aberrant splicing of the cyclin-dependent kinase-associated phosphatase, KAP, promotes glioblastoma invasion in a Cdc2-dependent manner. However, the mechanism by which this occurs is unknown. Here we show that miR-26a, which is often amplified in glioblastoma, promotes invasion in phosphatase and tensin homolog (PTEN)-competent and PTEN-deficient glioblastoma cells by directly downregulating KAP expression. Mechanistically, we find that KAP binds and activates ROCK2. Thus, RNA-mediated downregulation of KAP leads to decreased ROCK2 activity and this, in turn, increases Rac1-mediated invasion. In addition, the decrease in KAP expression activates the cyclin-dependent kinase, Cdk2, and this directly promotes invasion by increasing retinoblastoma phosphorylation, E2F-dependent Cdc2 expression and Cdc2-mediated inactivation of the actomyosin inhibitor, caldesmon. Importantly, glioblastoma cell invasion mediated by this pathway can be antagonized by Cdk2/Cdc2 inhibitors in vitro and in vivo. Thus, two distinct RNA-based mechanisms activate this novel KAP/ROCK2/Cdk2-dependent invasion pathway in glioblastoma.
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References
Mrugala MM . Advances and challenges in the treatment of glioblastoma: a clinician's perspective. Discov Med 2013; 15: 221–230.
Kwiatkowska A, Symons M . Signaling determinants of glioma cell invasion. Adv Exp Med Biol 2013; 986: 121–141.
Yu Y, Jiang X, Schoch BS, Carroll RS, Black PM, Johnson MD . Aberrant splicing of cyclin-dependent kinase-associated protein phosphatase KAP increases proliferation and migration in glioblastoma. Cancer Res 2007; 67: 130–138.
Hannon GJ, Casso D, Beach D . KAP: a dual specificity phosphatase that interacts with cyclin-dependent kinases. Proc Natl Acad Sci USA 1994; 91: 1731–1735.
Lin WR, Lai MW, Yeh CT . Cyclin-dependent kinase-associated protein phosphatase is overexpressed in alcohol-related hepatocellularcarcinoma and influences xenograft tumor growth. Oncol Rep 2013; 29: 903–910.
Yeh CT, Lu SC, Chen TC, Peng CY, Liaw YF . Aberrant transcripts of the cyclin-dependent kinase-associated protein phosphatase in hepatocellularcarcinoma. Cancer Res 2000; 60: 4697–4700.
Lee SW, Reimer CL, Fang L, Iruela-Arispe ML, Aaronson SA . Overexpression of kinase-associated phosphatase (KAP) in breast and prostate cancer and inhibition of the transformed phenotype by antisense KAP expression. Mol Cell Biol 2000; 20: 1723–1732.
Kim H, Huang W, Jiang X, Pennicooke B, Park PJ, Johnson MD . Integrative genome analysis reveals an oncomir/oncogene cluster regulating glioblastoma survivorship. Proc Natl Acad Sci USA 2010; 107: 2183–2188.
Huse JT, Brennan C, Hambardzumyan D, Wee B, Pena J, Rouhanifard SH et al. The PTEN-regulating microRNA miR-26a is amplified in high-grade glioma and facilitates gliomagenesis in vivo. Genes Dev 2009; 23: 1327–1337.
Liu B, Wu X, Liu B, Wang C, Liu Y, Zhou Q et al. MiR-26a enhances metastasis potential of lung cancer cells via AKT pathway by targeting PTEN. Biochim Biophys Acta 2012; 1822: 1692–1704.
Yang X, Liang L, Zhang XF, Jia HL, Qin Y, Zhu XC et al. MicroRNA-26a suppresses tumor growth and metastasis of human hepatocellularcarcinoma by targeting interleukin-6-Stat3 pathway. Hepatology 2013; 58: 158–170.
Yu L, Lu J, Zhang B, Liu X, Wang L, Li SY et al. miR-26a inhibits invasion and metastasis of nasopharyngealcancer by targeting EZH2. Oncol Lett 2013; 5: 1223–1228.
Wilkinson S, Paterson HF, Marshall CJ . Cdc42-MRCK and Rho-ROCK signalling cooperate in myosin phosphorylationand cell invasion. Nat Cell Biol 2005; 7: 255–261.
Salhia B, Rutten F, Nakada M, Beaudry C, Berens M, Kwan A et al. Inhibition of Rho-kinase affects astrocytoma morphology, motility, and invasion through activation of Rac1. Cancer Res 2005; 65: 8792–8800.
Li JQ, Miki H, Wu F, Saoo K, Nishioka M, Ohmori M et al. Cyclin A correlates with carcinogenesis and metastasis, and p27(kip1) correlates with lymphatic invasion, in colorectal neoplasms. Hum Pathol 2002; 33: 1006–1015.
Li JQ, Miki H, Ohmori M, Wu F, Funamoto Y . Expression of cyclin E and cyclin-dependent kinase 2 correlates with metastasis and prognosis in colorectal carcinoma. Hum Pathol 2001; 32: 945–953.
Pandithage R, Lilischkis R, Harting K, Wolf A, Jedamzik B, Luscher-Firzlaff J et al. The regulation of SIRT2 function by cyclin-dependent kinases affects cell motility. J Cell Biol 2008; 180: 915–929.
Berthet C, Kaldis P . Cdk2 and Cdk4 cooperatively control the expression of Cdc2. Cell Div 2006; 1: 10.
Han IS, Seo TB, Kim KH, Yoon JH, Yoon SJ, Namgung U . Cdc2-mediated Schwann cell migration during peripheral nerve regeneration. J Cell Sci 2007; 120: 246–255.
Satterwhite LL, Lohka MJ, Wilson KL, Scherson TY, Cisek LJ, Corden JL et al. Phosphorylationof myosin-II regulatory light chain by cyclin-p34cdc2: a mechanism for the timing of cytokinesis. J Cell Biol 1992; 118: 595–605.
Morrison DL, Sanghera JS, Stewart J, Sutherland C, Walsh MP, Pelech SL . Phosphorylationand activation of smooth muscle myosin light chainkinase by MAP kinase and cyclin-dependent kinase-1. Biochem Cell Biol 1996; 74: 549–557.
Knights MJ, Kyle S, Ismail A . Characteristic features of stem cells in glioblastomas: from cellular biology to genetics. Brain Pathol 2012; 22: 592–606.
Tawara S, Fukumoto Y, Shimokawa H . Effects of combined therapy with a Rho-kinase inhibitor and prostacyclin on monocrotaline-induced pulmonary hypertension in rats. J Cardiovasc Pharmacol 2007; 50: 195–200.
Yamaguchi H, Kasa M, Amano M, Kaibuchi K, Hakoshima T . Molecular mechanism for the regulation of rho-kinase by dimerization and its inhibition by fasudil. Structure 2006; 14: 589–600.
Jacobs M, Hayakawa K, Swenson L, Bellon S, Fleming M, Taslimi P et al. The structure of dimeric ROCK I reveals the mechanism for ligand selectivity. J Biol Chem 2006; 281: 260–268.
Amano M, Chihara K, Nakamura N, Kaneko T, Matsuura Y, Kaibuchi K . The COOH terminus of Rho-kinase negatively regulates rho-kinase activity. J Biol Chem 1999; 274: 32418–32424.
Ferretti R, Palumbo V, Di Savino A, Velasco S, Sbroggio M, Sportoletti P et al. Morgana/chp-1, a ROCK inhibitor involved in centrosome duplication and tumorigenesis. Dev Cell 2010; 18: 486–495.
Ma Z, Kanai M, Kawamura K, Kaibuchi K, Ye K, Fukasawa K . Interaction between ROCK II and nucleophosmin/B23 in the regulation of centrosome duplication. Mol Cell Biol 2006; 26: 9016–9034.
Yoneda A, Multhaupt HA, Couchman JR . The Rho kinases I and II regulate different aspects of myosin II activity. J Cell Biol 2005; 170: 443–453.
Corsino PE, Davis BJ, Norgaard PH, Parker NN, Law M, Dunn W et al. Mammary tumors initiated by constitutive Cdk2 activation contain an invasive basal-like component. Neoplasia 2008; 10: 1240–1252.
Bales E, Mills L, Milam N, McGahren-Murray M, Bandyopadhyay D, Chen D et al. The low molecular weight cyclin E isoforms augment angiogenesis and metastasis of human melanoma cells in vivo. Cancer Res 2005; 65: 692–697.
Joyce D, Bouzahzah B, Fu M, Albanese C, D'Amico M, Steer J et al. Integration of Rac-dependent regulation of cyclin D1 transcription through a nuclear factor-kappaB-dependent pathway. J Biol Chem 1999; 274: 25245–25249.
Nalepa G, Barnholtz-Sloan J, Enzor R, Dey D, He Y, Gehlhausen JR et al. The tumor suppressor CDKN3 controls mitosis. J Cell Biol 2013; 201: 997–1012.
Lowery DM, Clauser KR, Hjerrild M, Lim D, Alexander J, Kishi K et al. Proteomic screen defines the Polo-box domain interactome and identifies Rock2 as a Plk1 substrate. EMBO J 2007; 26: 2262–2273.
Wang HF, Takenaka K, Nakanishi A, Miki Y . BRCA2 and nucleophosmin coregulate centrosome amplification and form a complex with the Rho effector kinase ROCK2. Cancer Res 2011; 71: 68–77.
Kim TM, Huang W, Park R, Park PJ, Johnson MD . A developmental taxonomy of glioblastoma defined and maintained by MicroRNAs. Cancer Res 2011; 71: 3387–3399.
Yang HW, Menon LG, Black PM, Carroll RS, Johnson MD . SNAI2/Slug promotes growth and invasion in human gliomas. BMC Cancer 2010; 10: 301.
Acknowledgements
This work was supported by R01 NS062219 from the National Institute of Neurological Disorders and Stroke, a National Institutes of Health Director’s New Innovator Award (DP2OD002319) and a Brain Science Foundation Research Award to MDJ.
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Li, H., Jiang, X., Yu, Y. et al. KAP regulates ROCK2 and Cdk2 in an RNA-activated glioblastoma invasion pathway. Oncogene 34, 1432–1441 (2015). https://doi.org/10.1038/onc.2014.49
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DOI: https://doi.org/10.1038/onc.2014.49
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