RTVP-1 regulates glioma cell migration and invasion via interaction with N-WASP and hnRNPK

Glioblastoma (GBM) are characterized by increased invasion into the surrounding normal brain tissue. RTVP-1 is highly expressed in GBM and regulates the migration and invasion of glioma cells. To further study RTVP-1 effects we performed a pull-down assay using His-tagged RTVP-1 followed by mass spectrometry and found that RTVP-1 was associated with the actin polymerization regulator, N-WASP. This association was further validated by co-immunoprecipitation and FRET analysis. We found that RTVP-1 increased cell spreading, migration and invasion and these effects were at least partly mediated by N-WASP. Another protein which was found by the pull-down assay to interact with RTVP-1 is hnRNPK. This protein has been recently reported to associate with and to inhibit the effect of N-WASP on cell spreading. hnRNPK decreased cell migration, spreading and invasion in glioma cells. Using co-immunoprecipitation we validated the interactions of hnRNPK with N-WASP and RTVP-1 in glioma cells. In addition, we found that overexpression of RTVP-1 decreased the association of N-WASP and hnRNPK. In summary, we report that RTVP-1 regulates glioma cell spreading, migration and invasion and that these effects are mediated via interaction with N-WASP and by interfering with the inhibitory effect of hnRNPK on the function of this protein.

[1]  Li Jiang,et al.  Mammalian diaphanous-related formin 1 is required for motility and invadopodia formation in human U87 glioblastoma cells. , 2014, International journal of molecular medicine.

[2]  T. Mikkelsen,et al.  Related to testes-specific, vespid and pathogenesis protein-1 is regulated by methylation in glioblastoma , 2014, Oncology letters.

[3]  N. Mohandas,et al.  A 130-kDa Protein 4.1B Regulates Cell Adhesion, Spreading, and Migration of Mouse Embryo Fibroblasts by Influencing Actin Cytoskeleton Organization* , 2013, The Journal of Biological Chemistry.

[4]  H. Fathallah-Shaykh,et al.  c-Src and Neural Wiskott-Aldrich Syndrome Protein (N-WASP) Promote Low Oxygen-Induced Accelerated Brain Invasion by Gliomas , 2013, PloS one.

[5]  B. Baguley,et al.  Variable Expression of GLIPR1 Correlates with Invasive Potential in Melanoma Cells , 2013, Front. Oncol..

[6]  T. Mikkelsen,et al.  MicroRNA-137 is downregulated in glioblastoma and inhibits the stemness of glioma stem cells by targeting RTVP-1 , 2013, Oncotarget.

[7]  Nashi Widodo,et al.  Heterogeneous Nuclear Ribonucleoprotein K (hnRNP-K) Promotes Tumor Metastasis by Induction of Genes Involved in Extracellular Matrix, Cell Movement, and Angiogenesis , 2013, The Journal of Biological Chemistry.

[8]  P. Kloppenburg,et al.  CRN2 enhances the invasiveness of glioblastoma cells , 2013, Neuro-oncology.

[9]  T. Mikkelsen,et al.  MicroRNA-145 Is Downregulated in Glial Tumors and Regulates Glioma Cell Migration by Targeting Connective Tissue Growth Factor , 2013, PloS one.

[10]  F. Umansky,et al.  TRAIL conjugated to nanoparticles exhibits increased anti-tumor activities in glioma cells and glioma stem cells in vitro and in vivo. , 2013, Neuro-oncology.

[11]  H. Gil-Henn,et al.  Invadopodia: the leading force. , 2012, European journal of cell biology.

[12]  Amanda Y. Chan,et al.  The small GTPase RhoG mediates glioblastoma cell invasion , 2012, Molecular Cancer.

[13]  B. Shia,et al.  hnRNP Q Regulates Cdc42-Mediated Neuronal Morphogenesis , 2012, Molecular and Cellular Biology.

[14]  L. Parada,et al.  Malignant Glioma: Lessons from Genomics, Mouse Models, and Stem Cells , 2012, Cell.

[15]  I. Nakano,et al.  Glioma stem cells: their role in chemoresistance. , 2012, World neurosurgery.

[16]  R. Sarid,et al.  RTVP-1 expression is regulated by SRF downstream of protein kinase C and contributes to the effect of SRF on glioma cell migration. , 2011, Cellular signalling.

[17]  M. Tate,et al.  Biology of angiogenesis and invasion in glioma , 2009, Neurotherapeutics.

[18]  S. Kurisu,et al.  WASP and WAVE family proteins: Friends or foes in cancer invasion? , 2010, Cancer science.

[19]  H. Yamaguchi,et al.  Membrane lipids in invadopodia and podosomes: Key structures for cancer invasion and metastasis , 2010, Oncotarget.

[20]  Y. Ueyama,et al.  Proteomic analysis for nuclear proteins related to tumour malignant progression: a comparative proteomic study between malignant progressive cells and regressive cells. , 2010, Anticancer research.

[21]  Eric C. Holland,et al.  Targeting brain cancer: advances in the molecular pathology of malignant glioma and medulloblastoma , 2010, Nature Reviews Cancer.

[22]  O. Destaing,et al.  Actin machinery and mechanosensitivity in invadopodia, podosomes and focal adhesions , 2009, Journal of Cell Science.

[23]  A. Chakravarti,et al.  Molecular advances of brain tumors in radiation oncology. , 2009, Seminars in radiation oncology.

[24]  H. Fernando,et al.  WASP and WAVE proteins: vital intrinsic regulators of cell motility and their role in cancer (review). , 2009, International journal of molecular medicine.

[25]  S. Stylli,et al.  Invadopodia: At the cutting edge of tumour invasion , 2008, Journal of Clinical Neuroscience.

[26]  A. Luini,et al.  Multiple regulatory inputs converge on cortactin to control invadopodia biogenesis and extracellular matrix degradation , 2008, Journal of Cell Science.

[27]  L. Chin,et al.  Malignant astrocytic glioma: genetics, biology, and paths to treatment. , 2007, Genes & development.

[28]  T. Huang,et al.  Hypomethylation and aberrant expression of the glioma pathogenesis-related 1 gene in Wilms tumors. , 2007, Neoplasia.

[29]  K. Kinzler,et al.  A protocol for rapid generation of recombinant adenoviruses using the AdEasy system , 2007, Nature Protocols.

[30]  T. Haystead,et al.  PITK, a PP1 targeting subunit that modulates the phosphorylation of the transcriptional regulator hnRNP K. , 2006, Cellular signalling.

[31]  G I Murray,et al.  Heterogeneous nuclear ribonucleoprotein K is over expressed, aberrantly localised and is associated with poor prognosis in colorectal cancer , 2006, British Journal of Cancer.

[32]  J. Guan,et al.  Interaction of N-WASP with hnRNPK and Its Role in Filopodia Formation and Cell Spreading* , 2006, Journal of Biological Chemistry.

[33]  Angelo L. Vescovi,et al.  Brain tumour stem cells , 2006, Nature Reviews Cancer.

[34]  Y. Shoshan,et al.  Related to testes-specific, vespid, and pathogenesis protein-1 (RTVP-1) is overexpressed in gliomas and regulates the growth, survival, and invasion of glioma cells. , 2006, Cancer research.

[35]  Graeme I Murray,et al.  The roles of heterogeneous nuclear ribonucleoproteins in tumour development and progression. , 2006, Biochimica et biophysica acta.

[36]  A. Friedman,et al.  Chemotherapy and novel therapeutic approaches in malignant glioma. , 2005, Frontiers in bioscience : a journal and virtual library.

[37]  Shiro Suetsugu,et al.  Rac-WAVE2 signaling is involved in the invasive and metastatic phenotypes of murine melanoma cells , 2005, Oncogene.

[38]  C. Brodie,et al.  Protein kinase C-epsilon regulates the apoptosis and survival of glioma cells. , 2005, Cancer research.

[39]  R. Henkelman,et al.  Identification of human brain tumour initiating cells , 2004, Nature.

[40]  Nicole Rusk,et al.  Role of Synaptojanin 2 in Glioma Cell Migration and Invasion , 2004, Cancer Research.

[41]  G. Martin,et al.  Active Rho is localized to podosomes induced by oncogenic Src and is required for their assembly and function , 2004, The Journal of cell biology.

[42]  M. Mann,et al.  RNA and RNA Binding Proteins Participate in Early Stages of Cell Spreading through Spreading Initiation Centers , 2004, Cell.

[43]  L. Machesky,et al.  Signalling to actin assembly via the WASP (Wiskott-Aldrich syndrome protein)-family proteins and the Arp2/3 complex. , 2004, The Biochemical journal.

[44]  M. Ittmann,et al.  RTVP-1, a Tumor Suppressor Inactivated by Methylation in Prostate Cancer , 2004, Cancer Research.

[45]  Cynthia Hawkins,et al.  Identification of a cancer stem cell in human brain tumors. , 2003, Cancer research.

[46]  J. S. Rao,et al.  Molecular mechanisms of glioma invasiveness: the role of proteases , 2003, Nature Reviews Cancer.

[47]  M. Westphal,et al.  Cost of migration: invasion of malignant gliomas and implications for treatment. , 2003, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[48]  Y. Okada,et al.  The role of matrix metalloproteinases in glioma invasion. , 2003, Frontiers in bioscience : a journal and virtual library.

[49]  G. Reifenberger,et al.  The WHO Classification of Tumors of the Nervous System , 2002, Journal of neuropathology and experimental neurology.

[50]  Leszek Rychlewski,et al.  Heterogeneous Nuclear Ribonucleoprotein K Protein Associates with Multiple Mitochondrial Transcripts within the Organelle* , 2002, The Journal of Biological Chemistry.

[51]  E. Zamir,et al.  Molecular complexity and dynamics of cell-matrix adhesions. , 2001, Journal of cell science.

[52]  T. Takenawa,et al.  WASP and WAVE family proteins: key molecules for rapid rearrangement of cortical actin filaments and cell movement. , 2001, Journal of cell science.

[53]  M. Welch,et al.  The world according to Arp: regulation of actin nucleation by the Arp2/3 complex. , 1999, Trends in cell biology.

[54]  M. Kirschner,et al.  The Interaction between N-WASP and the Arp2/3 Complex Links Cdc42-Dependent Signals to Actin Assembly , 1999, Cell.

[55]  G. Dreyfuss,et al.  The K nuclear shuttling domain: a novel signal for nuclear import and nuclear export in the hnRNP K protein , 1997, The EMBO journal.

[56]  M. Israel,et al.  RTVP-1, a novel human gene with sequence similarity to genes of diverse species, is expressed in tumor cell lines of glial but not neuronal origin. , 1996, Gene.

[57]  J. Biggs,et al.  The human glioma pathogenesis-related protein is structurally related to plant pathogenesis-related proteins and its gene is expressed specifically in brain tumors. , 1995, Gene.