PGE2 Upregulates IL-8 Via P38MAPK-Dependent Dual-Activation of CHOP and C/EBP-β in Human Astrocytomas


  • Isabella Venza Department of Experimental Specialized Medical and Surgical and Odontostomatology Sciences, University of Messina, Messina, Italy
  • Maria Visalli Department ofClinical and Experimental Medicine, University of Messina, Messina, Ital
  • Rosaria Oteri Department ofClinical and Experimental Medicine, University of Messina, Messina, Ital
  • Federica Agliano Department of Clinical and Experimental Medicine, University of Messina, Messina, Italy
  • Silvia Morabito Department ofClinical and Experimental Medicine, University of Messina, Messina, Ital
  • Gerardo Caruso Neurosurgical Clinic,Department ofNeurosciences, University of Messina, Messina, Italy
  • Maria Caffo Neurosurgical Clinic,Department ofNeurosciences, University of Messina, Messina, Italy
  • Diana Teti Department ofClinical and Experimental Medicine, University of Messina, Messina, Ital



Astrocytoma, IL-8, PGE2, p38MAPK


We previously showed that in low- as well as in high-grade astrocytomas IL-8 overexpression is triggered by prostaglandin E2 (PGE2) through the upregulation of the transcription factors CCAAT/enhancer-binding protein-β (C/EBP-β) and C/EBP homologous protein (CHOP). Here we investigated the signal transduction pathways and the molecular mechanisms underlying the PGE2-dependent IL-8 gene expression in astrocytomas. Low- and high-grade PGE2-treated astrocytoma cells were transfected with wild-type and mutated IL-8 promoter constructs in the presence of various signal transduction pathway inhibitors, and cotransfected with transcription factor overexpressing plasmids or small-interfering RNAs. p38MAPK, C/EBP-β, and CHOP phosphorylation was analyzed by Western blotting. Electrophoretic mobility shift assay and chromatin immunoprecipitation evaluated the in vitro and in vivo binding of CHOP and C/EBP-β to IL-8 promoter. The results obtained allowed us to find out the signaling pathways triggered by PGE2 and responsible for the activation of the transcription factors involved in the overproduction of IL-8 by astrocytoma. Therefore, it can be argued that the inhibition of the PGE2 downstream pathways may represent a novel therapeutic approach for the treatment of patients with astrocytoma.


Wittekind C, Neid M. Cancer invasion and metastasis. Oncology 2005; 69: 14-6.

Van Meir EG. Cytokines and tumors of the central nervous system. Glia 1995; 15: 264-88.

Mandraffino G, Sardo MA, Riggio S, D'Ascola A, Loddo S, Alibrandi A, et al. Smoke exposure and circulating progenitor cells: evidence for modulation of antioxidant enzymes and cell count. Clin Biochem 2010; 43: 1436-42.

Koch AE, Polverini PJ, Kunkel SL, Harlow LA, Di Pietro LA, Elner VM, et al. Interleukin-8 as a macrophage-derived mediator of angiogenesis. Science1992; 258: 1798-801.

Wakabayashi K, Kambe F, Cao X, Murakami R, Mitsuyama H, Nagaya T, et al. Inhibitory effects of cyclosporine-A on calcium mobilization-dependent interleukin-8 expression and invasive potential of human glioblastoma U251MG cells. Oncogene 2004; 23: 6924-32.

Hoffmann E, Dittrich-Breiholz O, Holtmann H, Kracht M. Multiple control of interleukin-8 gene expression. J Leukoc Biol 2002; 72: 847-55.

Xie K. Interleukin-8 and human cancer biology. Cytokine Growth Factor Rev 2001; 12: 375-91.

Yamanaka R, Tanaka R, Yoshida S. Effects of irradiation on cytokine production in glioma cell lines. Neurol Med Chir 1993; 33: 744-8.

Desbaillets I, Diserens AC, Tribolet N, Hamou MF, Van Meir EG. Upregulation of interleukin 8 by oxygen-deprived cells in glioblastoma suggests a role in leukocyte activation, chemotaxis, and angiogenesis J Exp Med 1997; 186: 1201-12.

Kasahara T, Oda T, Hatake K, Akiyama M, Mukaida N, Matsushima K. Interleukin-8 and monocyte chemotactic protein-1 production by a human glioblastoma cell line, T98G in coculture with monocytes: involvement of monocyte-derived interleukin-1alpha. Eur Cytokine Netw 1998; 9: 47-55.

Palma C, Manzini S. Substance P induces secretion of immunomodulatory cytokines by human astrocytoma cells. J Neuroimmunol 1998; 81: 127-37.

Kehlen A, Thiele K, Riemann D, Rainov N, Langner J. Interleukin-17 stimulates the expression of IkappaB alpha mRNA and the secretion of IL-6 and IL-8 in glioblastoma cell lines J Neuroimmunol 1999; 101: 1-6.

Choi C, Kutsch O, Park J, Zhou T, Seol DW, Benveniste EN. Tumor necrosis factor-related apoptosis-inducing ligand induces caspase-dependent interleukin-8 expression and apoptosis in human astroglioma cells. Mol Cell Biol 2002; 22: 724-36.

Choi C, Xu X, Oh JW, Lee SJ, Gillespie GY, Park H, et al. Fas-induced expression of chemokines in human glioma cells: involvement of extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase. Cancer Res 2001; 61: 3084-91.

Venza M, Visalli M, Alafaci C, Caffo M, Caruso G, Salpietro FM, et al. Interleukin-8 (IL-8) overexpression in astrocytomas is induced by prostaglandin E2 (PGE2) and is associated to the transcription factors CCAAT enhancer binding protein-beta (C/EBP-β) and C/EBP Homologous Protein (CHOP). Neurosurgery 2011; 69: 713-21.

Venza I, Visalli M, Fortunato C, Ruggeri M, Ratone S, Caffo M, et al. PGE2 induces interleukin-8 derepression in human astrocytoma through coordinated DNA demethylation and histone hyperacetylation. Epigenetics 2012; 7: 1315-30.

Cucinotta M, Visalli M, Aguennouz M, Valenti A, Loddo S, Altucci L, Teti D. Regulation of interleukin-8 gene at a distinct site of its promoter by CCAAT enhancer-binding protein homologous protein in prostaglandin E2-treated human T cells. J Biol Chem 2008; 283: 29760-9.

Venza I, Cucinotta M, Caristi S, Mancuso G, Teti D. Transcriptional regulation of IL-8 by Staphylococcus aureus in human conjunctival cells involves activation of AP-1. Investig Ophthalmol Vis Sci 2007; 48: 270-6.

Wang XZ, Ron D. Stress-induced phosphorylation and activation of the transcription factor CHOP (GADD153) by p38 MAP kinase. Science 1996; 272: 1347-9.

Venza I, Cucinotta M, Visalli M, De Grazia G, Oliva S, Teti D. Pseudomonas aeruginosa induces interleukin-8 (IL-8) gene expression in human conjunctiva through the recruitment of both RelA and CCAAT/enhancer-binding protein beta to the IL-8 promoter. J Biol Chem 2009; 284: 4191-9.

Yao XH, Ping YF, Chen JH, Chen DL, Xu CP, Zheng J, et al. Production of angiogenic factors by human glioblastoma cells following activation of the G-protein coupled formylpeptide receptor FPR. J Neurooncol 2008; 86: 47-53.

Caffo M, Barresi V, Caruso G, Cutugno M, La Fata G, Venza M, et al. Innovative therapeutic strategies in the treatment of brain metastases. Int J Mol Sci 2013; 14: 2135-74.

Herlaar E, Brown Z. p38 MAPK signalling cascades in inflammatory disease. Mol Med Today 1999; 5: 439-47.

Ono K, Han J. The p38 signal transduction pathway: activation and function. Cell Signal 2000; 12: 1-13.

Kim SJ, Jeong HJ, Lee KM, Myung NY, An NH, Yang WM, et al. Epigallocatechin-3-gallate suppresses NF-kappaB activation and phosphorylation of p38 MAPK and JNK in human astrocytoma U373MG cells. J Nutr Biochem 2007; 18: 587-96.

Tokuda H, Kozawa O, Niwa M, Matsuno H, Kato K, Uematsu T. Mechanism of prostaglandin E2-stimulated heat shock protein 27 induction in osteoblast-like MC3T3-E1 cells. J Endocrinol 2002; 172: 271-81.

Fujino H, Xu W, Regan JW. Prostaglandin E2 induced functional expression of early growth response factor-1 by EP4, but not EP2, prostanoid receptors via the phosphatidylinositol 3-kinase and extracellular signal-regulated kinases. J Biol Chem 2003; 278: 12151-6.

Kobayashi Y, Mizoguchi T, Take I, Kurihara S, Udagawa N, Takahashi N. Prostaglandin E2 enhances osteoclastic differentiation of precursor cells through protein kinase A-dependent phosphorylation of TAK1. J Biol Chem 2005; 280: 11395-403.

Caristi S, Piraino G, Cucinotta M, Valenti A, Loddo S, Teti D. Prostaglandin E2 induces interleukin-8 gene transcription by activating C/EBP homologous protein in human T lymphocytes. J Biol Chem 2005; 280: 14433-42.

Delgado MA, Poschet JF, Deretic V. Nonclassical pathway of pseudomonas aeruginosa DNA-induced interleukin-8 secretion in cystic fibrosis airway epithelial cells. Infect Immun 2006; 74: 2975-84.

Matsusaka T, Fujikawa K, Nishio Y, Mukaida N, Mastushima K, Kishimoto T, et al. Transcription factors NF-IL6 and NF-kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. Proc Natl Acad Sci USA 1993; 90: 10193-7.

Kunsch C, Lang RK, Rosen CA, Shannon MF. Synergistic transcriptional activation of the IL-8 gene by NF-kappa B p65 (RelA) and NF-IL-6. J Immunol 1994; 153: 153-64.

Mukerjee R, Sawaya B E, Khalili K, Amini S. Association of p65 and C/EBPbeta with HIV-1 LTR modulates transcription of the viral promoter. J Cell Biochem 2007; 100: 1210-6.

Ruocco MR, Chen X, Ambrosino C, Dragonetti E, Liu W, Maliardo M, et al. Regulation of HIV-1 long terminal repeats by interaction of C/EBP(NF-IL6) and NF-kappaB/Rel transcription factors. J Biol Chem 1996; 271: 22479-86.

Mukaida N, Okamoto S, Ishikawa Y, Matsushima K. Molecular mechanism of interleukin-8 gene expression. J Leukoc Biol 1994; 56: 554-8.

Stein B, Baldwin AS Jr. Distinct mechanisms for regulation of the interleukin-8 gene involve synergism and cooperativity between C/EBP and NF-κB. Mol Cell Biol 1993; 13: 7191-8.

Vij N, Fang S, Zeitlin PL. Selective inhibition of endoplasmic reticulum-associated degradation rescues ΔF508-cystic fibrosis transmembrane regulator and suppresses interleukin-8 levels: Therapeutic implications. J Biol Chem 2006; 281: 17369-78.

Ron D, Habener JF. CHOP, a novel developmentally regulated nuclear protein that dimerizes with transcriptionfactors C/EBP and LAP and functions as a dominant negative inhibitor of gene transcription. Genes Dev 1992; 6: 439-53.

Shirakawa K, Maeda S, Gotoh T, Hayashi M, Shinomiya K, Ehata S, et al. CCAAT/enhancer-binding protein homologous protein (CHOP) regulates osteoblast differentiation. Mol Cell Biol 2006; 26: 6105-16.

John AE, Zhu YM, Brightling CE, Pang L, Knox AJ. Human airway smooth muscle cells from asthmatic individuals have CXCL8 hypersecretion due to increased NF-kappa B p65, C/EBP beta, and RNA polymerase II binding to the CXCL8 promoter. Immunol 2009; 183: 4682-92.

Liu S, Shi X, Bauer I, Günther J, Seyfert HM. Lingual antimicrobial peptide and IL-8 expression are oppositely regulated by the antagonistic effects of NF-κB p65 and C/EBPβ in mammary epithelial cells. Mol Immunol 2011; 48: 895-908.

Homma J, Yamanaka R, Yajima N, Tsuchiya N, Genkai N, Sano M, Tanaka R. Increased expression of CCAAT/enhancer binding protein beta correlates with prognosis in glioma patients. Oncol Rep 2006; 15: 595-601.

Ma J, Qiu Y, Yang L, Peng L, Xia Z, Hou LN, et al. Desipramine induces apoptosis in rat glioma cells via endoplasmic reticulum stress-dependent CHOP pathway. J Neurooncol 2010; 101: 41-8.

Pyrko P, Schönthal AH, Hofman FM, Chen TC, Lee AS. The unfolded protein response regulator GRP78/BiP as a novel target for increasing chemosensitivity in malignant gliomas. Cancer Res 2007; 67: 9809-16.

Nozell S, Laver T, Patel K, Benveniste EN. Mechanism of IFN-beta-mediated inhibition of IL-8 gene expression in astroglioma cells. J Immunol 2006; 177: 822-30.

Roebuck KA. Regulation of interleukin-8 gene expression. J Interferon Cytokine Res 1999; 19: 429-38.




How to Cite

Isabella Venza, Maria Visalli, Rosaria Oteri, Agliano, F. ., Silvia Morabito, Gerardo Caruso, Maria Caffo, & Diana Teti. (2014). PGE2 Upregulates IL-8 Via P38MAPK-Dependent Dual-Activation of CHOP and C/EBP-β in Human Astrocytomas. Journal of Analytical Oncology, 3(3),  146–158.




Most read articles by the same author(s)