Abrogation of PIK3CA or PIK3R1 reduces proliferation, migration, and invasion in glioblastoma multiforme cells

Glioblastoma multiforme (GBM) is a highly invasive and deadly brain tumor. Tumor cell invasion makes complete surgical resection impossible and reduces the efficacy of other therapies. Genome-wide analyses of mutations, copy-number changes, and expression patterns have provided new insights into genetic abnormalities common in GBM. We analyzed published data and identified the invasion and motility pathways most frequently altered in GBM. These were most notably the focal adhesion and integrin signaling, and extracellular matrix interactions pathways. We mapped alterations in each of these pathways and found that they included the catalytic PIK3CA and regulatory PIK3R1 subunit genes of the class IA PI3K. Knockdown of either of these genes separately in GBM cell lines by lentiviral-mediated shRNA expression resulted in decreased proliferation, migration, and invasion in all lines tested. FAK activity was reduced by knockdown of either PIK3CA or PIK3R1, and MMP2 levels were reduced by knockdown of PIK3R1. We conclude that PIK3R1, like PIK3CA, is a potential therapeutic target in GBM and that it also influences tumor cell growth and motility.

[1]  P. Burger,et al.  COMPUTERIZED TOMOGRAPHIC AND PATHOLOGIC STUDIES OF THE UNTREATED, QUIESCENT AND RECURRENT GLIOBLASTOMA MULTIFORME , 1983 .

[2]  W. Raub.  From the National Institutes of Health. , 1990, JAMA.

[3]  M. Berens,et al.  Determinants of human astrocytoma migration. , 1994, Cancer research.

[4]  H. Sönmez,et al.  Fibronectin and sialic acid levels in human meningiomas and gliomas. , 1995, Cancer letters.

[5]  S. Coons,et al.  Dichotomy of astrocytoma migration and proliferation , 1996, International journal of cancer.

[6]  D. Silbergeld,et al.  Isolation and characterization of human malignant glioma cells from histologically normal brain. , 1997, Journal of neurosurgery.

[7]  J. Backer,et al.  Regulation of the p85/p110 Phosphatidylinositol 3′-Kinase: Stabilization and Inhibition of the p110α Catalytic Subunit by the p85 Regulatory Subunit , 1998, Molecular and Cellular Biology.

[8]  J. Backer,et al.  Regulation of the p85/p110alpha phosphatidylinositol 3'-kinase. Distinct roles for the n-terminal and c-terminal SH2 domains. , 1998, The Journal of biological chemistry.

[9]  Mario Mellado,et al.  Role of the Pi3k Regulatory Subunit in the Control of Actin Organization and Cell Migration , 2000, The Journal of cell biology.

[10]  C. Damsky,et al.  FAK integrates growth-factor and integrin signals to promote cell migration , 2000, Nature Cell Biology.

[11]  J. Segall,et al.  N-terminal Domains of the Class IA Phosphoinositide 3-Kinase Regulatory Subunit Play a Role in Cytoskeletal but Not Mitogenic Signaling* , 2001, The Journal of Biological Chemistry.

[12]  A. Ljubimov,et al.  Overexpression of alpha4 chain-containing laminins in human glial tumors identified by gene microarray analysis. , 2001, Cancer research.

[13]  D. Bigner,et al.  EGF mutant receptor vIII as a molecular target in cancer therapy. , 2001, Endocrine-related cancer.

[14]  I. Campbell,et al.  The phosphatidylinositol 3'-kinase p85alpha gene is an oncogene in human ovarian and colon tumors. , 2001, Cancer research.

[15]  D. Durden,et al.  PTEN and phosphatidylinositol 3'-kinase inhibitors up-regulate p53 and block tumor-induced angiogenesis: evidence for an effect on the tumor and endothelial compartment. , 2003, Cancer research.

[16]  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.

[17]  T. Yamasaki,et al.  Growth inhibition of human malignant glioma cells induced by the PI3-K-specific inhibitor. , 2003, Journal of neurosurgery.

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

[19]  J. Ptak,et al.  High Frequency of Mutations of the PIK3CA Gene in Human Cancers , 2004, Science.

[20]  T. Klockgether,et al.  Molecular Analysis of the PTEN, TP53 and CDKN2A Tumor Suppressor Genes in Long-term Survivors of Glioblastoma Multiforme , 2000, Journal of Neuro-Oncology.

[21]  Wenjun Guo,et al.  Integrin signalling during tumour progression , 2004, Nature Reviews Molecular Cell Biology.

[22]  竹本 尚史.  Interaction between leukemic-cell VLA-4 and stromal fibronectin is a decisive factor for minimal residual disease of acute myelogenous leukemia , 2004 .

[23]  Y. Kanemura,et al.  Role of fibronectin-stimulated tumor cell migration in glioma invasion in vivo: clinical significance of fibronectin and fibronectin receptor expressed in human glioma tissues , 1998, Clinical & Experimental Metastasis.

[24]  C. Gondi,et al.  Downregulation of uPA, uPAR and MMP-9 using small, interfering, hairpin RNA (siRNA) inhibits glioma cell invasion, angiogenesis and tumor growth. , 2004, Neuron glia biology.

[25]  C. Livasy,et al.  Upregulation of focal adhesion kinase (FAK) expression in ductal carcinoma in situ (DCIS) is an early event in breast tumorigenesis , 2004, Breast Cancer Research and Treatment.

[26]  Catherine L Nutt,et al.  Genetic Alterations of Phosphoinositide 3‐kinase Subunit Genes in Human Glioblastomas , 2004, Brain pathology.

[27]  J. Condeelis,et al.  Breast Cancer Cells Isolated by Chemotaxis from Primary Tumors Show Increased Survival and Resistance to Chemotherapy , 2004, Cancer Research.

[28]  J. Nakamura,et al.  PKB/Akt mediates radiosensitization by the signaling inhibitor LY294002 in human malignant gliomas , 2005, Journal of Neuro-Oncology.

[29]  Carlo Rago,et al.  Mutant PIK3CA promotes cell growth and invasion of human cancer cells. , 2005, Cancer cell.

[30]  R. Kiss,et al.  Possible future issues in the treatment of glioblastomas: special emphasis on cell migration and the resistance of migrating glioblastoma cells to apoptosis. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[31]  Martin J. van den Bent,et al.  Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. , 2005, The New England journal of medicine.

[32]  K. Sekiguchi,et al.  Downregulation of laminin α4 chain expression inhibits glioma invasion in vitro and in vivo , 2005, International journal of cancer.

[33]  Gary L Gallia,et al.  PIK3CA Gene Mutations in Pediatric and Adult Glioblastoma Multiforme , 2006, Molecular Cancer Research.

[34]  D. Livant,et al.  Role of focal adhesion kinase and phosphatidylinositol 3'-kinase in integrin fibronectin receptor-mediated, matrix metalloproteinase-1-dependent invasion by metastatic prostate cancer cells. , 2006, Cancer research.

[35]  Marc-André Elsliger,et al.  Rare cancer-specific mutations in PIK3CA show gain of function , 2007, Proceedings of the National Academy of Sciences.

[36]  S. Frisch,et al.  Caspase-8 interacts with the p85 subunit of phosphatidylinositol 3-kinase to regulate cell adhesion and motility. , 2007, Cancer research.

[37]  B. Vanhaesebroeck,et al.  Class IA phosphoinositide 3-kinases are obligate p85-p110 heterodimers , 2007, Proceedings of the National Academy of Sciences.

[38]  Yuval Inbar,et al.  Mechanism of Two Classes of Cancer Mutations in the Phosphoinositide 3-Kinase Catalytic Subunit , 2007, Science.

[39]  M. Ewen,et al.  Wild-Type NRas and KRas Perform Distinct Functions during Transformation , 2007, Molecular and Cellular Biology.

[40]  Pingfu Fu,et al.  Activation of PI3K‐Akt signaling pathway promotes prostate cancer cell invasion , 2007, International journal of cancer.

[41]  D. Busam,et al.  An Integrated Genomic Analysis of Human Glioblastoma Multiforme , 2008, Science.

[42]  Li Zhao,et al.  Helical domain and kinase domain mutations in p110α of phosphatidylinositol 3-kinase induce gain of function by different mechanisms , 2008, Proceedings of the National Academy of Sciences.

[43]  J. Xiang,et al.  Disrupted RabGAP Function of the p85 Subunit of Phosphatidylinositol 3-Kinase Results in Cell Transformation* , 2008, Journal of Biological Chemistry.

[44]  S. Scherer,et al.  Erratum: Expression and mutation status of candidate kinases in multiple myeloma (Leukemia (2007) vol. 21 (1124-1127) 10.1038/sj.leu.2404612) , 2008 .

[45]  S. Scherer,et al.  Expression and mutation status of candidate kinases in multiple myeloma , 2008, Leukemia.

[46]  A. Kyritsis,et al.  Adenovirus-mediated transfer of siRNA against MMP-2 mRNA results in impaired invasion and tumor-induced angiogenesis, induces apoptosis in vitro and inhibits tumor growth in vivo in glioblastoma , 2008, Oncogene.

[47]  B. Peters,et al.  Somatic mutations in p85alpha promote tumorigenesis through class IA PI3K activation. , 2009, Cancer cell.

[48]  N. Stanietsky,et al.  The interaction of TIGIT with PVR and PVRL2 inhibits human NK cell cytotoxicity , 2009, Proceedings of the National Academy of Sciences.

[49]  Leyla Isik,et al.  Cancer-specific high-throughput annotation of somatic mutations: computational prediction of driver missense mutations. , 2009, Cancer research.

[50]  Andrés J. García,et al.  Focal adhesion kinase modulates cell adhesion strengthening via integrin activation. , 2009, Molecular biology of the cell.

[51]  G. Riggins,et al.  A survey of glioblastoma genomic amplifications and deletions , 2009, Journal of Neuro-Oncology.

[52]  Jin-Wu Nam,et al.  miR-29 miRNAs activate p53 by targeting p85α and CDC42 , 2009, Nature Structural &Molecular Biology.

[53]  S. Qiu,et al.  The effect of epidermal growth factor receptor variant III on glioma cell migration by stimulating ERK phosphorylation through the focal adhesion kinase signaling pathway. , 2010, Archives of biochemistry and biophysics.

[54]  Jing Xu,et al.  Tie2/TEK Modulates the Interaction of Glioma and Brain Tumor Stem Cells with Endothelial Cells and Promotes an Invasive Phenotype , 2010, Oncotarget.

[55]  K. Kinzler,et al.  PI3Kα Inhibitors That Inhibit Metastasis , 2010, Oncotarget.

[56]  P. Vogt,et al.  Cancer-derived mutations in the regulatory subunit p85α of phosphoinositide 3-kinase function through the catalytic subunit p110α , 2010, Proceedings of the National Academy of Sciences.

[57]  L. Zawel.  P3Ka: A Driver of Tumor Metastasis? , 2010, Oncotarget.

[58]  M. D. Chamberlain,et al.  Direct positive regulation of PTEN by the p85 subunit of phosphatidylinositol 3-kinase , 2010, Proceedings of the National Academy of Sciences.

[59]  R. Kaneva,et al.  Rare mutations in the PIK3CA gene contribute to aggressive endometrial cancer. , 2010, DNA and cell biology.

[60]  N. Yoo,et al.  Somatic mutation of PIK3R1 gene is rare in common human cancers , 2010, Acta oncologica.

[61]  S. Sengupta,et al.  Short hairpin RNA-mediated fibronectin knockdown delays tumor growth in a mouse glioma model. , 2010, Neoplasia.

[62]  Ws. Rasband.  ImageJ, U.S. National Institutes of Health, Bethesda, Maryland, USA , 2011 .

[63]  F. Lang,et al.  Glioblastoma angiogenesis and tumor cell invasiveness are differentially regulated by β8 integrin. , 2011, Cancer research.

[64]  G. van der Pluijm,et al.  Integrin αv expression is required for the acquisition of a metastatic stem/progenitor cell phenotype in human prostate cancer. , 2011, The American journal of pathology.

[65]  D. Sgroi,et al.  PIK3R1 (p85α) is somatically mutated at high frequency in primary endometrial cancer. , 2011, Cancer research.