Cyr61 Is Overexpressed in Gliomas and Involved in Integrin-Linked Kinase-Mediated Akt and β-Catenin-TCF/Lef Signaling Pathways

Cyr61 is a member of the CCN family of growth factors; these proteins are secreted and can act as ligands of distinct integrins. We show that Cyr61 can enhance tumorigenicity of glioma cells acting through activated integrin-linked kinase (ILK) to stimulate β-catenin-TCF/Lef and Akt signaling pathways. Overexpression of Cyr61 occurred in highly tumorigenic glioma cell lines and in 68% of the most malignant glioblastoma multiforme brain tumors. Forced expression of Cyr61 in U343 glioma cells accelerated their growth in liquid culture, enhanced their anchorage-independent proliferation in soft agar, and significantly increased their ability to form large, vascularized tumors in nude mice. Overexpression of Cyr61 in the U343 cells led to the up-regulation of distinct integrins, including β1 and ανβ3, which have been shown to interact with Cyr61 and ILK. The activity of ILK was increased dramatically in these cells. Overexpression of Cyr61 also resulted in the phosphorylation of glycogen synthase kinase-3β and accumulation and nuclear translocation of β-catenin, leading to activation of the β-catenin-TCF/Lef-1 signaling pathway. Furthermore, forced expression of Cyr61 in the glioma cells activated phosphatidylinositol 3′-kinase pathway, resulting in prominent phosphorylation of Akt and the antiapoptotic protein Bad. Cyr61 appears to stimulate several signaling pathways in the development of gliomas.

[1]  C. Ackerley,et al.  Tracking the invasiveness of human astrocytoma cells by using green fluorescent protein in an organotypical brain slice model. , 2001, Journal of neurosurgery.

[2]  F. Bertrand,et al.  Insulin and IGF-1 stimulate the β-catenin pathway through two signalling cascades involving GSK-3β inhibition and Ras activation , 2001, Oncogene.

[3]  L. Lau,et al.  Activation-dependent Adhesion of Human Platelets to Cyr61 and Fisp12/Mouse Connective Tissue Growth Factor Is Mediated through Integrin αIIbβ3 * , 1999, The Journal of Biological Chemistry.

[4]  Gary R. Grotendorst,et al.  Expression of connective tissue growth factor mRNA in the fibrous stroma of mammary tumors. , 1997, The international journal of biochemistry & cell biology.

[5]  P. Cohen,et al.  Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B , 1995, Nature.

[6]  Michael Kühl,et al.  Functional interaction of β-catenin with the transcription factor LEF-1 , 1996, Nature.

[7]  S. Shurtleff,et al.  Overexpression of mouse D-type cyclins accelerates G1 phase in rodent fibroblasts. , 1993, Genes & development.

[8]  L. Donehower,et al.  The Integrin-linked Kinase Regulates the Cyclin D1 Gene through Glycogen Synthase Kinase 3β and cAMP-responsive Element-binding Protein-dependent Pathways* , 2000, The Journal of Biological Chemistry.

[9]  Eric C. Holland,et al.  Combined activation of Ras and Akt in neural progenitors induces glioblastoma formation in mice , 2000, Nature Genetics.

[10]  M. Hüfner,et al.  The human analog of murine cystein rich protein 61 [correction of 16] is a 1alpha,25-dihydroxyvitamin D3 responsive immediate early gene in human fetal osteoblasts: regulation by cytokines, growth factors, and serum. , 1998, Endocrinology.

[11]  W. Cavenee,et al.  Genetics and malignant progression of human brain tumours. , 1995, Cancer surveys.

[12]  W F Bodmer,et al.  Target genes of beta-catenin-T cell-factor/lymphoid-enhancer-factor signaling in human colorectal carcinomas. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[13]  N. M. Khazenzon,et al.  Gene expression abnormalities in human glial tumors identified by gene array. , 2001, International journal of oncology.

[14]  M. Hüfner,et al.  The Human Analog of Murine Cystein Rich Protein 16 Is a 1α,25-Dihydroxyvitamin D3 Responsive Immediate Early Gene in Human Fetal Osteoblasts: Regulation by Cytokines, Growth Factors, and Serum. , 1998, Endocrinology.

[15]  Dianqing Wu,et al.  Suppression of Glycogen Synthase Kinase Activity Is Not Sufficient for Leukemia Enhancer Factor-1 Activation* , 1999, The Journal of Biological Chemistry.

[16]  R. Hynes Targeted mutations in cell adhesion genes: what have we learned from them? , 1996, Developmental biology.

[17]  F. McCormick,et al.  Differential Regulation of Glycogen Synthase Kinase 3β by Insulin and Wnt Signaling* , 2000, The Journal of Biological Chemistry.

[18]  L. Lau,et al.  Adhesion of Human Umbilical Vein Endothelial Cells to the Immediate-Early Gene Product Cyr61 Is Mediated through Integrin αvβ3 * , 1998, The Journal of Biological Chemistry.

[19]  K. Kinzler,et al.  Constitutive Transcriptional Activation by a β-Catenin-Tcf Complex in APC−/− Colon Carcinoma , 1997, Science.

[20]  T. V. Kolesnikova,et al.  CYR61, a product of a growth factor-inducible immediate early gene, promotes angiogenesis and tumor growth. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Elizabeth Yang,et al.  Serine Phosphorylation of Death Agonist BAD in Response to Survival Factor Results in Binding to 14-3-3 Not BCL-XL , 1996, Cell.

[22]  J W Smith,et al.  Integrin activation controls metastasis in human breast cancer. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Michael W. Peterson,et al.  Lipopolysaccharide Activates Akt in Human Alveolar Macrophages Resulting in Nuclear Accumulation and Transcriptional Activity of β-Catenin1 , 2001, The Journal of Immunology.

[24]  L. Strong,et al.  Structural analysis of the human nov proto-oncogene and expression in Wilms tumor. , 1994, Oncogene.

[25]  D. Botstein,et al.  WISP genes are members of the connective tissue growth factor family that are up-regulated in wnt-1-transformed cells and aberrantly expressed in human colon tumors. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[26]  R. Bravo,et al.  Complexity of the early genetic response to growth factors in mouse fibroblasts , 1988, Molecular and cellular biology.

[27]  M. E. Ruaro,et al.  Gas6 Induces Growth, β-Catenin Stabilization, and T-Cell Factor Transcriptional Activation in Contact-Inhibited C57 Mammary Cells , 2001, Molecular and Cellular Biology.

[28]  J. Brugge,et al.  Phosphatidylinositol 3-kinase is required for integrin-stimulated AKT and Raf-1/mitogen-activated protein kinase pathway activation , 1997, Molecular and cellular biology.

[29]  S. Fukumoto,et al.  Akt Participation in the Wnt Signaling Pathway through Dishevelled* , 2001, The Journal of Biological Chemistry.

[30]  S. Dedhar,et al.  Inhibition of integrin-linked kinase (ILK) suppresses activation of protein kinase B/Akt and induces cell cycle arrest and apoptosis of PTEN-mutant prostate cancer cells. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[31]  P. Polakis Wnt signaling and cancer. , 2000, Genes & development.

[32]  J. Woodgett,et al.  Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Diehl,et al.  Wnt1 and MEK1 Cooperate to Promote Cyclin D1 Accumulation and Cellular Transformation* , 2000, The Journal of Biological Chemistry.

[34]  L. Lau,et al.  Adhesion of Human Skin Fibroblasts to Cyr61 Is Mediated through Integrin α6β1 and Cell Surface Heparan Sulfate Proteoglycans* , 2000, The Journal of Biological Chemistry.

[35]  B. Herrmann,et al.  Nuclear localization of β-catenin by interaction with transcription factor LEF-1 , 1996, Mechanisms of Development.

[36]  L. D.,et al.  Brain tumors , 2005, Psychiatric Quarterly.

[37]  O. Kallioniemi,et al.  Identification of differentially expressed genes in human gliomas by DNA microarray and tissue chip techniques. , 2000, Cancer research.

[38]  M. Mayhaus,et al.  Muscarinic Acetylcholine Receptors Induce the Expression of the Immediate Early Growth Regulatory Gene CYR61 * , 2000, The Journal of Biological Chemistry.

[39]  R. Winneker,et al.  Cyr61, a Member of the CCN Family, Is Required for MCF-7 Cell Proliferation: Regulation by 17β-Estradiol and Overexpression in Human Breast Cancer. , 2001, Endocrinology.

[40]  H. Hanafusa,et al.  Transcriptional down regulation of the nov proto-oncogene in fibroblasts transformed by p60v-src , 1996, Molecular and cellular biology.

[41]  Y. Nakabeppu,et al.  Genomic response to growth factors. , 1988, Cold Spring Harbor symposia on quantitative biology.

[42]  M. Berger,et al.  Akt pathway activation converts anaplastic astrocytoma to glioblastoma multiforme in a human astrocyte model of glioma. , 2001, Cancer research.

[43]  S. Dedhar,et al.  Integrin-linked Protein Kinase Regulates Fibronectin Matrix Assembly, E-cadherin Expression, and Tumorigenicity* , 1998, The Journal of Biological Chemistry.

[44]  A. Levine,et al.  WISP-1 attenuates p53-mediated apoptosis in response to DNA damage through activation of the Akt kinase. , 2002, Genes & development.

[45]  L. Lau,et al.  Expression of a set of growth-related immediate early genes in BALB/c 3T3 cells: coordinate regulation with c-fos or c-myc. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[46]  Gary R. Grotendorst,et al.  Connective tissue growth factor: a cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10 , 1991, The Journal of cell biology.

[47]  Gordon Mills,et al.  Protein kinase B (PKB/Akt) activity is elevated in glioblastoma cells due to mutation of the tumor suppressor PTEN/MMAC , 1998, Current Biology.

[48]  S. Dedhar,et al.  Inhibition of integrin linked kinase (ILK) suppresses β-catenin-Lef/Tcf-dependent transcription and expression of the E-cadherin repressor, snail, in APC−/− human colon carcinoma cells , 2001, Oncogene.

[49]  L. Lau,et al.  Identification of a set of genes expressed during the G0/G1 transition of cultured mouse cells. , 1985, The EMBO journal.

[50]  David E. Misek,et al.  Distinctive molecular profiles of high-grade and low-grade gliomas based on oligonucleotide microarray analysis. , 2001, Cancer research.

[51]  A. Giaccia,et al.  Inhibition of the anti-apoptotic PI(3)K/Akt/Bad pathway by stress. , 1998, Genes & development.

[52]  S. R. Datta,et al.  Akt Phosphorylation of BAD Couples Survival Signals to the Cell-Intrinsic Death Machinery , 1997, Cell.

[53]  L. Lau,et al.  CYR61 Stimulates Human Skin Fibroblast Migration through Integrin αvβ5 and Enhances Mitogenesis through Integrin αvβ3, Independent of Its Carboxyl-terminal Domain* , 2001, The Journal of Biological Chemistry.

[54]  R. Elashoff,et al.  Elevated levels of connective tissue growth factor, WISP-1, and CYR61 in primary breast cancers associated with more advanced features. , 2001, Cancer research.

[55]  W. Cavenee,et al.  Accumulation of genetic defects during astrocytoma progression , 1992, Cancer.

[56]  H. Hameister,et al.  Expression and differential regulation of connective tissue growth factor in pancreatic cancer cells , 1999, Oncogene.

[57]  A. Levine,et al.  WISP-1 is a Wnt-1- and β-catenin-responsive oncogene , 2000, Genes & Development.

[58]  L. Strong,et al.  Structural analysis of the human nov proto-oncogene and expression in Wilms tumors , 1994 .

[59]  V. Joliot,et al.  Proviral rearrangements and overexpression of a new cellular gene (nov) in myeloblastosis-associated virus type 1-induced nephroblastomas , 1992, Molecular and cellular biology.

[60]  A. Sparks,et al.  Identification of c-MYC as a target of the APC pathway. , 1998, Science.

[61]  S. Dedhar,et al.  Cell adhesion and the integrin-linked kinase regulate the LEF-1 and beta-catenin signaling pathways. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[62]  K Satyamoorthy,et al.  N-cadherin-mediated intercellular interactions promote survival and migration of melanoma cells. , 2001, Cancer research.

[63]  Kakinuma,et al.  Expression of fibrogenic cytokines in desmoplastic malignant melanoma , 1998, The British journal of dermatology.

[64]  Hans Clevers,et al.  XTcf-3 Transcription Factor Mediates β-Catenin-Induced Axis Formation in Xenopus Embryos , 1996, Cell.

[65]  J. Lakins,et al.  Expression and function of CYR61, an angiogenic factor, in breast cancer cell lines and tumor biopsies. , 2000, Cancer research.

[66]  Carl W. Miller,et al.  Breast Cancer , 2001, The Journal of Biological Chemistry.

[67]  S. Dedhar,et al.  Regulation of cell adhesion and anchorage-dependent growth by a new β1-integrin-linked protein kinase , 1996, Nature.

[68]  Asim Khwaja,et al.  Matrix adhesion and Ras transformation both activate a phosphoinositide 3‐OH kinase and protein kinase B/Akt cellular survival pathway , 1997, The EMBO journal.

[69]  L. Peso,et al.  Interleukin-3-induced phosphorylation of BAD through the protein kinase Akt. , 1997, Science.

[70]  C. Ackerley,et al.  Co-expression of nestin and vimentin intermediate filaments in invasive human astrocytoma cells , 1999, International Journal of Developmental Neuroscience.

[71]  Yuqin Wang,et al.  Disintegrin Interaction with αvβ3Integrin on Human Umbilical Vein Endothelial Cells: Expression of Ligand-Induced Binding Site on β3Subunit , 1996 .

[72]  L. Lau,et al.  Activation-dependent adhesion of human platelets to Cyr61 and Fisp12/mouse connective tissue growth factor is mediated through integrin alpha(IIb)beta(3). , 1999, The Journal of biological chemistry.