Expression and function of Toll-like receptors in peripheral blood mononuclear cells from patients with ovarian cancer.

Inflammation has been implicated in the initiation and progression of ovarian cancer (OC), the underlying mechanisms of which are still unclear. We hypothesized that the abnormal expression of Toll-like receptors (TLRs), which were potential activators of nuclear factor-kappa B p65 (NF-κB p65), could promote inflammation and tumorigenesis in OC. In this study, we characterized the expression of TLRs in peripheral blood mononuclear cells (PBMCs) and found TLR2 and TLR6 mRNAs levels to be higher in PBMCs from OC patients than in those from benign disease (BC) or healthy normal controls (NC). Flow cytometry analysis showed that TLR1, TLR2 and TLR6 were highly expressed in monocytes from OC patients, but not in those from control subjects. Consistently, inflammatory cytokines interleukin (IL)-1β and IL-6 were up-regulated in PBMCs from OC patients upon stimulation with Pam3CSK4 (TLR1 ligand) and HKLM (TLR2 ligand), compared with unstimulated PBMCs. Stimulation of PBMCs with TLR ligands led to activation of downstream signaling molecules in TLRs (MyD88, TRAF6, TANK, NF-κB p65 and p-NF-κB p65). We also discovered that SK-OV-3-secreted factors were potent PBMCs activators, leading to the production of IL-1β, IL-6 and IL-8 through activation of TLRs and downstream signaling molecules in PBMCs. Before coculturing with SK-OV-3, pretreatment of THP-1 cells or PBMCs with monoclonal antibodies against TLR1, TLR2 or TLR6 inhibited the production of IL-1β and IL-6 and activation of MyD88, TRAF6, TANK, NF-κB p65 and p-NF-κB p65. Our results provided new evidence that TLR1, TLR2 and TLR6 signaling was linked with inflammation in OC microenvironment. Similar content being viewed by others Abbreviations - BC: - Benign disease control - EOC: - Epithelial ovarian cancer - FBS: - Fetal bovine serum - HRP: - Horseradish peroxidase - IL: - Interleukin - MyD88: - Myeloid differentiation factor 88 - NC: - Healthy normal controls - NF-κB: - Nuclear factor-kappa B - OC: - Ovarian cancer - PAMPs: - Pathogen-associated molecular patterns - PBMCs: - Peripheral blood mononuclear cells - p-NF-κB: - Phospho-Nuclear factor-kappa B - RT-PCR: - Real-time PCR - TANK: - TRAF family member-associated NF-κB activator - TLRs: - Toll-like receptors - TNFα: - Tumor necrosis factorα - TRAF6: - Tumor necrosis factor receptor-associated factor 6 - TRAIL: - TNF-related apoptosis-inducing ligand

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This work was supported by National Natural Science Foundation of China (81272324, 81371894), Key Laboratory for Medicine of Jiangsu Province of China (No. XK201114), a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions. The funders had no role in the study design, data collection and analysis, decision to publish or preparation of the manuscript. Conflict of interest The authors declare no financial or commercial conflict of interest. Author information Authors and Affiliations Corresponding authors Additional information Xiaojie Zhang, Juan Xu and Xing Ke have contributed equally to this work. Rights and permissions About this article Cite this article Zhang, X., Xu, J., Ke, X. et al. Expression and function of Toll-like receptors in peripheral blood mononuclear cells from patients with ovarian cancer. Cancer Immunol Immunother 64, 275–286 (2015). https://doi.org/10.1007/s00262-014-1632-x Received: Accepted: Published: Issue date: DOI: https://doi.org/10.1007/s00262-014-1632-x