PMID- 27791989 OWN - NLM STAT- MEDLINE DCOM- 20180222 LR - 20210103 IS - 1949-2553 (Electronic) IS - 1949-2553 (Linking) VI - 7 IP - 50 DP - 2016 Dec 13 TI - Stimulation of the toll-like receptor 3 promotes metabolic reprogramming in head and neck carcinoma cells. PG - 82580-82593 LID - 10.18632/oncotarget.12892 [doi] AB - In this study, a possible link between the innate immune recognition receptor TLR3 and metabolic reprogramming in Head and Neck carcinoma (HNC) cells was investigated. The effects of TLR3 stimulation/knock-down were assessed under several culture conditions in 4 HNC cell-lines by cell growth assays, targeted metabolomics, and glycolysis assays based on time-resolved analysis of proton release (Seahorse analyzer). The stimulation of TLR3 by its synthetic agonist Poly(A:U) resulted in a faster growth of HNC cells under low foetal calf serum conditions. Targeted analysis of glucose metabolism pathways demonstrated a tendency towards a shift from tricarboxylic acid cycle (Krebs cycle) to glycolysis and anabolic reactions in cells treated with Poly(A:U). Glycolysis assays confirmed that TLR3 stimulation enhanced the capacity of malignant cells to switch from oxidative phosphorylation to extra-mitochondrial glycolysis. We found evidence that HIF-1alpha is involved in this process: addition of the TLR3 agonist resulted in a higher cell concentration of the HIF-1alpha protein, even in normoxia, whereas knocking-down TLR3 resulted in a lower concentration, even in hypoxia. Finally, we assessed TLR3 expression by immunohistochemistry in a series of 7 HNSCC specimens and found that TLR3 was detected at higher levels in tumors displaying a hypoxic staining pattern. Overall, our results demonstrate that TLR3 stimulation induces the Warburg effect in HNC cells in vitro, and suggest that TLR3 may play a role in tumor adaptation to hypoxia. FAU - Veyrat, Mathieu AU - Veyrat M AD - University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France. FAU - Durand, Sylvere AU - Durand S AD - Equipe 11 Labelisee par la Ligue Nationale Contre le Cancer, INSERM U1138, Centre de Recherche des Cordeliers, Paris, France. AD - Metabolomics and Molecular Cell Biology Platforms, Gustave Roussy, Villejuif, France. FAU - Classe, Marion AU - Classe M AD - Department of Pathology, Lariboisiere Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France. FAU - Glavan, Tanja Matijevic AU - Glavan TM AD - Division of Molecular Medicine, Rudjer Boskovic Institute, Zagreb, Croatia. FAU - Oker, Natalie AU - Oker N AD - University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France. AD - Department of Head and Neck surgery, Lariboisiere Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France. FAU - Kapetanakis, Nikiforos-Ioannis AU - Kapetanakis NI AD - University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France. FAU - Jiang, Xiaojun AU - Jiang X AD - University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France. FAU - Gelin, Aurore AU - Gelin A AD - University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France. FAU - Herman, Philippe AU - Herman P AD - Department of Head and Neck surgery, Lariboisiere Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France. FAU - Casiraghi, Odile AU - Casiraghi O AD - Department of Biopathology, Gustave Roussy, Villejuif, France. FAU - Zagzag, David AU - Zagzag D AD - Department of Neuropathology, New York University School of Medicine, New York, NY, USA. FAU - Enot, David AU - Enot D AD - Equipe 11 Labelisee par la Ligue Nationale Contre le Cancer, INSERM U1138, Centre de Recherche des Cordeliers, Paris, France. AD - Metabolomics and Molecular Cell Biology Platforms, Gustave Roussy, Villejuif, France. FAU - Busson, Pierre AU - Busson P AD - University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France. FAU - Verillaud, Benjamin AU - Verillaud B AD - University Paris-Sud (Paris 11), CNRS-UMR 8126, Gustave Roussy, Villejuif, France. AD - Department of Head and Neck surgery, Lariboisiere Hospital, AP-HP, University Paris-Diderot Paris 7, Paris, France. LA - eng PT - Journal Article PL - United States TA - Oncotarget JT - Oncotarget JID - 101532965 RN - 0 (Antigens, Neoplasm) RN - 0 (HIF1A protein, human) RN - 0 (Hypoxia-Inducible Factor 1, alpha Subunit) RN - 0 (TLR3 protein, human) RN - 0 (Toll-Like Receptor 3) RN - 24936-38-7 (Poly A-U) RN - EC 4.2.1.1 (CA9 protein, human) RN - EC 4.2.1.1 (Carbonic Anhydrase IX) SB - IM MH - Antigens, Neoplasm/metabolism MH - Carbonic Anhydrase IX/metabolism MH - Carcinoma, Squamous Cell/genetics/*metabolism/pathology MH - Cell Line, Tumor MH - Cell Proliferation/drug effects MH - Cellular Reprogramming/*drug effects MH - Citric Acid Cycle/drug effects MH - Energy Metabolism/*drug effects MH - Glycolysis/drug effects MH - Head and Neck Neoplasms/genetics/*metabolism/pathology MH - Humans MH - Hypoxia-Inducible Factor 1, alpha Subunit/metabolism MH - Oxidative Phosphorylation/drug effects MH - Poly A-U/*pharmacology MH - RNA Interference MH - Signal Transduction/drug effects MH - Squamous Cell Carcinoma of Head and Neck MH - Time Factors MH - Toll-Like Receptor 3/*agonists/genetics/metabolism MH - Transfection MH - Tumor Hypoxia PMC - PMC5347715 OTO - NOTNLM OT - HIF OT - innate immunity OT - metabolomics OT - toll-like receptor 3 OT - warburg effect COIS- CONFLICTS OF INTEREST All authors certify that they have no conflicts of interest. EDAT- 2016/10/30 06:00 MHDA- 2018/02/23 06:00 PMCR- 2016/12/13 CRDT- 2016/10/30 06:00 PHST- 2015/02/12 00:00 [received] PHST- 2016/10/19 00:00 [accepted] PHST- 2016/10/30 06:00 [pubmed] PHST- 2018/02/23 06:00 [medline] PHST- 2016/10/30 06:00 [entrez] PHST- 2016/12/13 00:00 [pmc-release] AID - 12892 [pii] AID - 10.18632/oncotarget.12892 [doi] PST - ppublish SO - Oncotarget. 2016 Dec 13;7(50):82580-82593. doi: 10.18632/oncotarget.12892.