The survival for patients with high-grade glioma is poor, and only a limited number of patients respond to the therapy. The aim of this study was to analyze the significance of using p38 MAPK phosphorylation as a prognostic marker in high-grade glioma patients and as a therapeutic target in combination chemotherapy with vandetanib. p38 MAPK phosphorylation was analyzed with immunohistochemistry in 90 high-grade glioma patients. Correlation between p38 MAPK phosphorylation and overall survival was analyzed with Mann–Whitney U test analysis. The effects on survival of glioblastoma cells of combining vandetanib with the p38 MAPK inhibitor SB 203580 were analyzed in vitro with the median-effect method with the fluorometric microculture cytotoxicity assay. Two patients had phosphorylated p38 MAPK in both the cytoplasm and nucleus, and these two presented with worse survival than patients with no detectable p38 MAPK phosphorylation or phosphorylated p38 MAPK only in the nucleus. This was true for both high-grade glioma patients (WHO grade III and IV, n = 90, difference in median survival: 6.1 months, 95 % CI [0.20, 23], p = 0.039) and for the subgroup with glioblastoma patients (WHO grade IV, n = 70, difference in median survival: 6.1 months, 95 % CI [0.066, 23], p = 0.043). The combination of vandetanib and the p38 MAPK inhibitor SB 203580 had synergistic effects on cell survival for glioblastoma-derived cells in vitro. In conclusion, p38 MAPK phosphorylation may be a prognostic marker for high-grade glioma patients, and vandetanib combined with a p38 MAPK inhibitor may be useful combination chemotherapy for glioma patients.
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The authors would like to thank Dr. H Hedman, Umea University, the UCSF Tissue Bank, and Dr. JS Guillamo for providing us with the glioma cell lines used in the experiments. We would also like to thank Dr. E. Freyhult at Bioinformatics Infrastructure for Life Sciences for statistical support. In addition, we would also like to express our gratitude for the financial support from the Cancer Foundation at Gavle Hospital, the Research Fund at the Department of Oncology, Uppsala University Hospital, the Swedish Cancer Society, the Swedish Research Council, and the Knut and Alice Wallenberg Foundation.
Conflict of interest
The authors declare that we have no conflict of interest.
Dose-effect curves (A) and median-effect plots (B) of vandetanib combined with SB 203580. The combination was tested in a vandetanib sensitive (left panel) and resistant cell line (right panel). fa = affected fraction, fu = unaffected fraction. (TIFF 2987 kb)
van den Bent MJ, Dubbink HJ, Sanson M, van der Lee-Haarloo CR, Hegi M, Jeuken JWM, et al. MGMT promoter methylation is prognostic but not predictive for outcome to adjuvant PCV chemotherapy in anaplastic oligodendroglial tumors: a report from EORTC Brain Tumor Group Study 26951. J Clin Oncol. 2009;27(35):5881–6.PubMedCrossRefGoogle Scholar
Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352(10):987–96.PubMedCrossRefGoogle Scholar
Stupp R, Tonn JC, Brada M, Pentheroudakis G, Group EGW. High-grade malignant glioma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2010;21(Suppl 5):v190–3.PubMedCrossRefGoogle Scholar
Wagner EF, Nebreda AR. Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 2009;9(8):537–49.PubMedCrossRefGoogle Scholar
Demuth T, Reavie LB, Rennert JL, Nakada M, Nakada S, Hoelzinger DB, et al. MAP-ing glioma invasion: mitogen-activated protein kinase kinase 3 and p38 drive glioma invasion and progression and predict patient survival. Mol Cancer Ther. 2007;6(4):1212–22.PubMedCrossRefGoogle Scholar
Greenberg AK, Basu S, Hu J, Yie TA, Tchou-Wong KM, Rom WN, et al. Selective p38 activation in human non-small cell lung cancer. Am J Respir Cell Mol Biol. 2002;26(5):558–64.PubMedCrossRefGoogle Scholar
Esteva FJ, Sahin AA, Smith TL, Yang Y, Pusztai L, Nahta R, et al. Prognostic significance of phosphorylated P38 mitogen-activated protein kinase and HER-2 expression in lymph node-positive breast carcinoma. Cancer. 2004;100(3):499–506.PubMedCrossRefGoogle Scholar
Handra-Luca A, Lesty C, Hammel P, Sauvanet A, Rebours V, Martin A, et al. Biological and prognostic relevance of mitogen-activated protein kinases in pancreatic adenocarcinoma. Pancreas. 2012;41(3):416–21.PubMedCrossRefGoogle Scholar
Huang W, Deng B, Wang RW, Tan QY, He Y, Jiang YG, et al. BCAR1 protein plays important roles in carcinogenesis and predicts poor prognosis in non-small-cell lung cancer. PLoS One. 2012;7(4):e36124.PubMedCrossRefGoogle Scholar
Herbst RS, Sun Y, Eberhardt WE, Germonpre P, Saijo N, Zhou C, et al. Vandetanib plus docetaxel versus docetaxel as second-line treatment for patients with advanced non-small-cell lung cancer (ZODIAC): a double-blind, randomised, phase 3 trial. Lancet Oncol. 2010;11(7):619–26.PubMedCrossRefGoogle Scholar
Damiano V, Melisi D, Bianco C, Raben D, Caputo R, Fontanini G, et al. Cooperative antitumor effect of multitargeted kinase inhibitor ZD6474 and ionizing radiation in glioblastoma. Clin Cancer Res. 2005;11(15):5639–44.PubMedCrossRefGoogle Scholar
Drappatz J, Norden AD, Wong ET, Doherty LM, Lafrankie DC, Ciampa A, et al. Phase I study of vandetanib with radiotherapy and temozolomide for newly diagnosed glioblastoma. Int J Radiat Oncol Biol Phys. 2010;78(1):85–90.PubMedCrossRefGoogle Scholar
Paillas S, Boissiere F, Bibeau F, Denouel A, Mollevi C, Causse A, et al. Targeting the p38 MAPK pathway inhibits irinotecan resistance in colon adenocarcinoma. Cancer Res. 2011;71(3):1041–9.PubMedCrossRefGoogle Scholar
Guo X, Ma N, Wang J, Song J, Bu X, Cheng Y, et al. Increased p38-MAPK is responsible for chemotherapy resistance in human gastric cancer cells. BMC Cancer. 2008;8:375.PubMedCrossRefGoogle Scholar
Hirose Y, Katayama M, Stokoe D, Haas-Kogan DA, Berger MS, Pieper RO. The p38 mitogen-activated protein kinase pathway links the DNA mismatch repair system to the G2 checkpoint and to resistance to chemotherapeutic DNA-methylating agents. Mol Cell Biol. 2003;23(22):8306–15.PubMedCrossRefGoogle Scholar
Yoshino Y, Aoyagi M, Tamaki M, Duan L, Morimoto T, Ohno K. Activation of p38 MAPK and/or JNK contributes to increased levels of VEGF secretion in human malignant glioma cells. Int J Oncol. 2006;29(4):981–7.PubMedGoogle Scholar
Gee E, Milkiewicz M, Haas TL. p38 MAPK activity is stimulated by vascular endothelial growth factor receptor 2 activation and is essential for shear stress-induced angiogenesis. J Cell Physiol. 2010;222(1):120–6.PubMedCrossRefGoogle Scholar
Yilmaz A, Kliche S, Mayr-Beyrle U, Fellbrich G, Waltenberger J. p38 MAPK inhibition is critically involved in VEGFR-2-mediated endothelial cell survival. Biochem Biophys Res Commun. 2003;306(3):730–6.PubMedCrossRefGoogle Scholar
Mueller KL, Powell K, Madden JM, Eblen ST, Boerner JL. EGFR tyrosine 845 phosphorylation-dependent proliferation and transformation of breast cancer cells require activation of p38 MAPK. Transl Oncol. 2012;5(5):327–34.PubMedGoogle Scholar
Kampf C, Olsson I, Ryberg U, Sjostedt E, Ponten F. Production of tissue microarrays, immunohistochemistry staining and digitalization within the human protein atlas. J Vis Exp. 2012; (63):e3620. doi:10.3791/3620.
Lindhagen E, Nygren P, Larsson R. The fluorometric microculture cytotoxicity assay. Nat Protoc. 2008;3(8):1364–9.PubMedCrossRefGoogle Scholar
Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 1984;22:27–55.PubMedCrossRefGoogle Scholar
Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res. 2010;70(2):440–6.PubMedCrossRefGoogle Scholar
Uhlen M, Oksvold P, Fagerberg L, Lundberg E, Jonasson K, Forsberg M, et al. Towards a knowledge-based Human Protein Atlas. Nat Biotechnol. 2010;28(12):1248–50.PubMedCrossRefGoogle Scholar