The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) regulates the production of proangiogenic molecules by myeloma cells and suppresses hypoxia-inducible factor-1 alpha (HIF-1alpha) activity: involvement in myeloma-induced angiogenesis.

Angiogenesis has a critical role in the pathophysiology of multiple myeloma (MM); however, the molecular mechanisms underlying this process are not completely elucidated. The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) has been recently implicated in solid tumors as a repressor of angiogenesis. In this study, we found that ING4 expression in MM cells was correlated with the expression of the proangiogenic molecules interleukin-8 (IL-8) and osteopontin (OPN). Moreover, we demonstrate that ING4 suppression in MM cells up-regulated IL-8 and OPN, increasing the hypoxia inducible factor-1alpha (HIF-1alpha) activity and its target gene NIP-3 expression in hypoxic condition. In turn, we show that the inhibition of HIF-1alpha by siRNA suppressed IL-8 and OPN production by MM cells under hypoxia. A direct interaction between ING4 and the HIF prolyl hydroxylase 2 (HPH-2) was also demonstrated. Finally, we show that ING4 suppression in MM cells significantly increased vessel formation in vitro, blunted by blocking IL-8 or OPN. These in vitro observations were confirmed in vivo by finding that MM patients with high IL-8 production and microvascular density (MVD) have significantly lower ING4 levels compared with those with low IL-8 and MVD. Our data indicate that ING4 exerts an inhibitory effect on the production of proangiogenic molecules and consequently on MM-induced angiogenesis.

[1]  M. Pierotti,et al.  Detection of novel mRNA splice variants of human ING4 tumor suppressor gene , 2007, Oncogene.

[2]  Shaji K. Kumar,et al.  Cytokine and chemokine profiles in multiple myeloma; significance of stromal interaction and correlation of IL-8 production with disease progression. , 2007, Leukemia research.

[3]  M. Simon,et al.  Hypoxia-inducible factors: central regulators of the tumor phenotype. , 2007, Current opinion in genetics & development.

[4]  G. Casey,et al.  Breast cancer metastasis suppressor 1 (BRMS1) inhibits osteopontin transcription by abrogating NF-κB activation , 2007, Molecular Cancer.

[5]  V. Kalra,et al.  A Novel Role of Hypoxia-Inducible Factor in Cobalt Chloride- and Hypoxia-Mediated Expression of IL-8 Chemokine in Human Endothelial Cells12 , 2006, The Journal of Immunology.

[6]  C. Harris,et al.  Novel Splice Variants of ING4 and Their Possible Roles in the Regulation of Cell Growth and Motility* , 2006, Journal of Biological Chemistry.

[7]  G. Roodman,et al.  Chemokines in multiple myeloma. , 2006, Experimental hematology.

[8]  John Crowley,et al.  The molecular classification of multiple myeloma. , 2006, Blood.

[9]  J. Pouysségur,et al.  The role of the hypoxia-inducible factor in tumor metabolism growth and invasion. , 2006, Bulletin du cancer.

[10]  G. Semenza,et al.  Regulation of angiogenesis by hypoxia-inducible factor 1. , 2006, Critical reviews in oncology/hematology.

[11]  M. Simon,et al.  Regulation of angiogenesis by hypoxia and hypoxia-inducible factors. , 2006, Current topics in developmental biology.

[12]  S. Colla,et al.  Human myeloma cells express the bone regulating gene Runx2/Cbfa1 and produce osteopontin that is involved in angiogenesis in multiple myeloma patients , 2005, Leukemia.

[13]  P. Sutphin,et al.  Hypoxia upregulates osteopontin expression in NIH-3T3 cells via a Ras-activated enhancer , 2005, Oncogene.

[14]  Y. Ninomiya,et al.  Frequent deletion and down-regulation of ING4, a candidate tumor suppressor gene at 12p13, in head and neck squamous cell carcinomas. , 2005, Gene.

[15]  A. Ozer,et al.  Regulation of HIF by Prolyl hydroxylases: Recruitment of the Candidate Tumor Suppressor Protein ING4 , 2005, Cell cycle.

[16]  T. Tsuruo,et al.  Blockade of the stromal cell-derived factor-1/CXCR4 axis attenuates in vivo tumor growth by inhibiting angiogenesis in a vascular endothelial growth factor-independent manner. , 2005, Cancer research.

[17]  Ze-Guang Han,et al.  Nuclear localization signal of ING4 plays a key role in its binding to p53. , 2005, Biochemical and biophysical research communications.

[18]  Abdullah Ozer,et al.  The candidate tumor suppressor ING4 represses activation of the hypoxia inducible factor (HIF). , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Erwin G. Van Meir,et al.  The role of interleukin-8 and its receptors in gliomagenesis and tumoral angiogenesis. , 2005, Neuro-oncology.

[20]  M. Toi,et al.  Thymidine phosphorylase (platelet-derived endothelial-cell growth factor) in cancer biology and treatment. , 2005, The Lancet. Oncology.

[21]  Suwon Kim.  HuntING4 New Tumor Suppressors , 2005, Cell cycle.

[22]  S. Colla,et al.  Lack of receptor activator of nuclear factor-kB ligand (RANKL) expression and functional production by human multiple myeloma cells. , 2005, Haematologica.

[23]  Jeremy J. W. Chen,et al.  The role of interleukin-8 in cancer cells and microenvironment interaction. , 2005, Frontiers in bioscience : a journal and virtual library.

[24]  M. Ridder,et al.  Role of the hypoxic bone marrow microenvironment in 5T2MM murine myeloma tumor progression. , 2005, Haematologica.

[25]  K. Chin,et al.  A screen for genes that suppress loss of contact inhibition: identification of ING4 as a candidate tumor suppressor gene in human cancer. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[26]  A. Harris,et al.  Differential Function of the Prolyl Hydroxylases PHD1, PHD2, and PHD3 in the Regulation of Hypoxia-inducible Factor* , 2004, Journal of Biological Chemistry.

[27]  Shaji K. Kumar,et al.  Bone marrow angiogenic ability and expression of angiogenic cytokines in myeloma: evidence favoring loss of marrow angiogenesis inhibitory activity with disease progression. , 2004, Blood.

[28]  P. Ongusaha,et al.  HB-EGF Is a Potent Inducer of Tumor Growth and Angiogenesis , 2004, Cancer Research.

[29]  E. Stathopoulos,et al.  The relation between bone marrow angiogenesis and the proliferation index Ki-67 in multiple myeloma , 2004, Journal of Clinical Pathology.

[30]  R. Jain,et al.  The candidate tumour suppressor protein ING4 regulates brain tumour growth and angiogenesis , 2004, Nature.

[31]  Gonzalez,et al.  Insights into the multistep transformation of MGUS to myeloma using microarray expression analysis. , 2003, Blood.

[32]  F. Watzinger,et al.  Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using ‘real-time’ quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) – a Europe against cancer program , 2003, Leukemia.

[33]  J. Pouysségur,et al.  HIF prolyl‐hydroxylase 2 is the key oxygen sensor setting low steady‐state levels of HIF‐1α in normoxia , 2003, The EMBO journal.

[34]  K. Nishio,et al.  Osteopontin overproduced by tumor cells acts as a potent angiogenic factor contributing to tumor growth. , 2003, Cancer letters.

[35]  R. Bataille,et al.  Proangiogenic properties of human myeloma cells: production of angiopoietin-1 and its potential relationship to myeloma-induced angiogenesis. , 2003, Blood.

[36]  Koh Miura,et al.  p29ING4 and p28ING5 bind to p53 and p300, and enhance p53 activity. , 2003, Cancer research.

[37]  R. Lai,et al.  Proliferation, apoptosis, and intratumoral vascularity in multiple myeloma: correlation with the clinical stage and cytological grade. , 2002, Journal of clinical pathology.

[38]  R. Fonseca,et al.  Bone marrow angiogenesis in 400 patients with monoclonal gammopathy of undetermined significance, multiple myeloma, and primary amyloidosis. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[39]  N. Saijo,et al.  Osteopontin induces angiogenesis of murine neuroblastoma cells in mice , 2002, International journal of cancer.

[40]  John Crowley,et al.  Global gene expression profiling of multiple myeloma, monoclonal gammopathy of undetermined significance, and normal bone marrow plasma cells. , 2002, Blood.

[41]  Yung‐Chie Lee,et al.  Aberrant p53 expression correlates with expression of vascular endothelial growth factor mRNA and interleukin-8 mRNA and neoangiogenesis in non-small-cell lung cancer. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[42]  N. Munshi,et al.  Increased bone marrow microvessel density in newly diagnosed multiple myeloma carries a poor prognosis. , 2001, Seminars in oncology.

[43]  R. Ransohoff,et al.  TNF-α mediates SDF-1α–induced NF-κB activation and cytotoxic effects in primary astrocytes , 2001 .

[44]  M. Mollenauer,et al.  Endogenous CD28 expressed on myeloma cells up-regulates interleukin-8 production: implications for multiple myeloma progression. , 2001, Blood.

[45]  M. Burdick,et al.  CXC chemokines in angiogenesis , 2000, Journal of leukocyte biology.

[46]  W. Berdel,et al.  Vascular endothelial growth factor and interleukin-6 in paracrine tumor-stromal cell interactions in multiple myeloma. , 2000, Blood.

[47]  D. Ribatti,et al.  Bone marrow neovascularization, plasma cell angiogenic potential, and matrix metalloproteinase-2 secretion parallel progression of human multiple myeloma. , 1999, Blood.

[48]  Luigi Naldini,et al.  Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo , 1997, Nature Biotechnology.

[49]  S. Schwartz,et al.  Osteopontin promotes vascular cell adhesion and spreading and is chemotactic for smooth muscle cells in vitro. , 1994, Circulation research.