Aryl hydrocarbon receptor inhibition downregulates the TGF-β/Smad pathway in human glioblastoma cells

The dioxin/aryl hydrocarbon receptor (AhR) is a transcription factor, which has been attributed a role in human cancerogenesis, cell cycle progression and transforming growth factor-β (TGF-β) signaling. As TGF-β is an important mediator of the malignant phenotype of human gliomas, we studied AhR expression and function in glioma cells. AhR was not only expressed in glioma cells in vitro, but was also detected in human gliomas in vivo by immunohistochemistry, with a predominantly nuclear staining in glioblastomas. The AhR agonist, 3-methylcholanthrene, induced AhR nuclear translocation and upregulated mRNA levels of the AhR target gene, cytochrome P450 1A1 (CYP1A1). Conversely, pharmacological inhibition of AhR using the novel AhR antagonist, CH-223191, or AhR gene silencing using small interfering RNA showed that constitutive AhR activity positively controls TGF-β1, TGF-β2 and latent TGF-β-binding protein-1 protein levels in malignant glioma cells. Moreover, antagonism of AhR reduced clonogenic survival and invasiveness of glioma cells. In contrast, AhR regulates TGF-β signaling negatively in non-neoplastic astrocytes. Thus, the pathogenesis of glioma formation may involve altered AhR regulation of the TGF-β/Smad pathway, and AhR may represent a promising target for the treatment of human malignant gliomas and other diseases associated with pathological TGF-β activity.

[1]  Y. Fujii‐Kuriyama,et al.  Benzo[a]pyrene carcinogenicity is lost in mice lacking the aryl hydrocarbon receptor. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[2]  C. Elferink.  Aryl hydrocarbon receptor-mediated cell cycle control. , 2003, Progress in cell cycle research.

[3]  J. Kononen,et al.  Tissue microarrays for high-throughput molecular profiling of tumor specimens , 1998, Nature Medicine.

[4]  Xiaoqing Chang,et al.  LTBP‐1 blockade in dioxin receptor‐null mouse embryo fibroblasts decreases TGF‐β activity: Role of extracellular proteases plasmin and elastase , 2006, Journal of cellular biochemistry.

[5]  K. Miyazono,et al.  Regulation of TGF-beta signaling and its roles in progression of tumors. , 2003, Cancer science.

[6]  J. Buer,et al.  The aryl hydrocarbon receptor links TH17-cell-mediated autoimmunity to environmental toxins , 2008, Nature.

[7]  Yishi Jin,et al.  The AHR-1 aryl hydrocarbon receptor and its co-factor the AHA-1 aryl hydrocarbon receptor nuclear translocator specify GABAergic neuron cell fate in C. elegans , 2004, Development.

[8]  C. Tohyama,et al.  A Constitutively Active Arylhydrocarbon Receptor Induces Growth Inhibition of Jurkat T Cells through Changes in the Expression of Genes Related to Apoptosis and Cell Cycle Arrest* , 2004, Journal of Biological Chemistry.

[9]  S. Kress,et al.  Functional analysis of the human cytochrome P4501A1 (CYP1A1) gene enhancer. , 1998, European journal of biochemistry.

[10]  M. Weller,et al.  SD-208, a Novel Transforming Growth Factor β Receptor I Kinase Inhibitor, Inhibits Growth and Invasiveness and Enhances Immunogenicity of Murine and Human Glioma Cells In vitro and In vivo , 2004, Cancer Research.

[11]  Sun-Hee Kim,et al.  Novel Compound 2-Methyl-2H-pyrazole-3-carboxylic Acid (2-methyl-4-o-tolylazo-phenyl)-amide (CH-223191) Prevents 2,3,7,8-TCDD-Induced Toxicity by Antagonizing the Aryl Hydrocarbon Receptor , 2006, Molecular Pharmacology.

[12]  A. Puga,et al.  Aryl hydrocarbon receptor, cell cycle regulation, toxicity, and tumorigenesis , 2005, Journal of cellular biochemistry.

[13]  K. Miyazono,et al.  Regulation of TGF‐β signaling and its roles in progression of tumors , 2003 .

[14]  M. Denison,et al.  Activation of the aryl hydrocarbon receptor by structurally diverse exogenous and endogenous chemicals. , 2003, Annual review of pharmacology and toxicology.

[15]  A Poland,et al.  2,3,7,8-tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons: examination of the mechanism of toxicity. , 1982, Annual review of pharmacology and toxicology.

[16]  Thai D. Nguyen,et al.  WT1 Expression Distinguishes Astrocytic Tumor Cells from Normal and Reactive Astrocytes , 2008, Brain pathology.

[17]  Y. Fujii‐Kuriyama,et al.  Functional role of AhR in the expression of toxic effects by TCDD. , 2003, Biochimica et biophysica acta.

[18]  Y. Fujii‐Kuriyama,et al.  Characterization of xenobiotic responsive elements upstream from the drug-metabolizing cytochrome P-450c gene: a similarity to glucocorticoid regulatory elements. , 1987, Nucleic acids research.

[19]  A. Hanberg,et al.  A constitutively active dioxin/aryl hydrocarbon receptor induces stomach tumors , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[20]  S. Anderson,et al.  Integration of Smad and Forkhead Pathways in the Control of Neuroepithelial and Glioblastoma Cell Proliferation , 2004, Cell.

[21]  B. Scheithauer,et al.  The 2007 WHO classification of tumours of the central nervous system , 2007, Acta Neuropathologica.

[22]  P. Fernández-Salguero,et al.  Liver portal fibrosis in dioxin receptor‐null mice that overexpress the latent transforming growth factor‐β‐binding protein‐1 , 2004, International journal of experimental pathology.

[23]  L. Poellinger,et al.  Purification of the DNA binding form of dioxin receptor. Role of the Arnt cofactor in regulation of dioxin receptor function. , 1994, The Journal of biological chemistry.

[24]  S. Mulero-Navarro,et al.  Overexpression of latent transforming growth factor-β binding protein 1 (LTBP-1) in dioxin receptor-null mouse embryo fibroblasts , 2004, Journal of Cell Science.

[25]  Alvaro Puga,et al.  Ligand-Independent Regulation of Transforming Growth Factor β1 Expression and Cell Cycle Progression by the Aryl Hydrocarbon Receptor , 2007, Molecular and Cellular Biology.

[26]  Paola Pisani,et al.  Genetic Pathways to Glioblastoma , 2004, Cancer Research.

[27]  C. Tohyama,et al.  Modulation of oestrogen receptor signalling by association with the activated dioxin receptor , 2003, Nature.

[28]  D. Louis.  WHO classification of tumours of the central nervous system , 2007 .

[29]  T. Haarmann-Stemmann,et al.  Growth factors, cytokines and their receptors as downstream targets of arylhydrocarbon receptor (AhR) signaling pathways. , 2009, Biochemical pharmacology.

[30]  R S McCuskey,et al.  Portosystemic shunting and persistent fetal vascular structures in aryl hydrocarbon receptor-deficient mice. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[31]  Michael Platten,et al.  Glioma Cell Invasion: Regulation of Metalloproteinase Activity by TGF-β , 2001, Journal of Neuro-Oncology.

[32]  Y. Fujii‐Kuriyama,et al.  Intrinsic AhR function underlies cross-talk of dioxins with sex hormone signalings. , 2008, Biochemical and biophysical research communications.

[33]  L. Poellinger,et al.  A Constitutively Active Dioxin/Aryl Hydrocarbon Receptor Promotes Hepatocarcinogenesis in Mice , 2004, Cancer Research.

[34]  M. Weller,et al.  RNA Interference Targeting Transforming Growth Factor-β Enhances NKG2D-Mediated Antiglioma Immune Response, Inhibits Glioma Cell Migration and Invasiveness, and Abrogates Tumorigenicity In vivo , 2004, Cancer Research.

[35]  A. Poland,et al.  Response of murine epidermis to 2,3,7,8-tetrachlorodibenzo-p-dioxin: Interaction of the Ah and hr loci , 1982, Cell.

[36]  E. Glover,et al.  Stereospecific, high affinity binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin by hepatic cytosol. Evidence that the binding species is receptor for induction of aryl hydrocarbon hydroxylase. , 1976, The Journal of biological chemistry.

[37]  Y. Fujii‐Kuriyama,et al.  Dioxin receptor is a ligand-dependent E3 ubiquitin ligase , 2007, Nature.

[38]  Alejandra Bruna,et al.  High TGFbeta-Smad activity confers poor prognosis in glioma patients and promotes cell proliferation depending on the methylation of the PDGF-B gene. , 2007, Cancer cell.

[39]  L. Poellinger,et al.  Definition of a Dioxin Receptor Mutant That Is a Constitutive Activator of Transcription , 2001, The Journal of Biological Chemistry.

[40]  Robert Walgate,et al.  Proliferation , 1985, Nature.

[41]  G. Albertin,et al.  Similar sequence-free amplification of human glyceraldehyde-3-phosphate dehydrogenase for real time RT-PCR applications. , 2005, Molecular and cellular probes.

[42]  M. Weller,et al.  Modulation of TGF‐β activity by latent TGF‐β‐binding protein 1 in human malignant glioma cells , 2009, International journal of cancer.

[43]  Fred Hirsch,et al.  The aryl hydrocarbon receptor repressor is a putative tumor suppressor gene in multiple human cancers. , 2008, The Journal of clinical investigation.

[44]  Adriano Fontana,et al.  The failure of current immunotherapy for malignant glioma. Tumor-derived TGF-β, T-cell apoptosis, and the immune privilege of the brain , 1995, Brain Research Reviews.

[45]  Y. Fujii‐Kuriyama,et al.  [Xenobiotics and transcriptional regulation]. , 2003, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[46]  K. Kinzler,et al.  Human Smad3 and Smad4 are sequence-specific transcription activators. , 1998, Molecular cell.

[47]  J. Hogenesch,et al.  The PAS superfamily: sensors of environmental and developmental signals. , 2000, Annual review of pharmacology and toxicology.

[48]  The aryl hydrocarbon receptor agonist 3,3',4,4',5-pentachlorobiphenyl induces distinct patterns of gene expression between hepatoma and glioma cells: chromatin remodeling as a mechanism for selective effects. , 2007, Neurotoxicology.

[49]  A. Fornace,et al.  The involvement of aryl hydrocarbon receptor in the activation of transforming growth factor-beta and apoptosis. , 1998, Molecular pharmacology.

[50]  P. Harper,et al.  Regulating the regulator: factors that control levels and activity of the aryl hydrocarbon receptor. , 2006, Biochemical pharmacology.

[51]  P. Harper,et al.  Cell-specific regulation of human aryl hydrocarbon receptor expression by transforming growth factor-beta(1). , 2001, Molecular pharmacology.

[52]  H. Weiner,et al.  Control of Treg and TH17 cell differentiation by the aryl hydrocarbon receptor , 2008, Nature.