The tumor-associated inflammatory microenvironment may play a pivotal role in epithelial ovarian cancer (EOC) carcinogenesis and outcomes, but a detailed profile in patient-derived tumors is needed. Here, we investigated the expression of TLR4- and MyD88-associated markers in tumors from over 500 EOC patients using immunohistochemical staining. We demonstrate that high expression of TLR4 and MyD88 predicts poorer overall survival in patients with EOC; most likely, this is due to their association with serous histology and features of high tumor burden and aggressiveness, including stage, grade, and ascites at surgery. Combined TLR4 and MyD88 expression appears to serve as an independent risk factor for shortened survival time, even after covariate adjustment (both moderate HR 1.1 [95 % CI 0.7–1.8], both strong HR 2.1 [95 % CI 1.1–3.8], both weak as referent; p = 0.027). We reveal that in EOC tissues with elevated expression of both TLR4 and MyD88 and activated NF-κB signaling pathway, expression of hsp60, hsp70, beta 2 defensin, and HMGB1 are also enriched. In total, these results suggest that activation of TLR4/MyD88/NF-κB signaling by endogenous ligands may contribute to an inflammatory microenvironment that drives a more aggressive phenotype with poorer clinical outcome in EOC patients.
Epithelial ovarian cancer Toll-like receptor four (TLR4) Myeloid differentiation primary response gene eighty-eight (MyD88) Endogenous ligands NF-κB signaling pathway
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This work was supported by the Mayo Clinic SPORE in Ovarian Cancer, P50 CA136393, and R01 CA122443.
Compliance with ethical standards
We declare that all experiments were performed in accordance with the current law of USA. The study was approved by the Institutional Review Board (IRB) of Mayo Clinic in Rochester. Patients provided written informed consent and permission for active follow-up.
Conflicts of interest
The authors declare that they have no conflict of interest
Representative images for endogenous ligands, TLR4, and MyD88 Scoring was based on the intensities of nuclear and cytoplasmic staining in the majority of cells. In order to conserve figure space, markers are shown grouped by the scoring scheme used in final analyses; in some instances, this represents combination of score categories: none/weak v moderate/strong (hsp60); none/weak v moderate (gp96, beta 2 defensin); none v weak/moderate (fibrinogen, heparan sulfate); none v weak v moderate (hsp70, HMGB1); and none/weak v moderate v strong (TLR4 and MyD88). (GIF 347 kb)
Representative images for NF-κB signaling pathway and target gene In order to conserve figure space, markers are shown grouped by the scoring scheme used in final analyses; in some instances, this represents combination of score categories: none v weak v moderate intensity of nuclear and cytoplasmic staining in the majority of cells (IKKβ, IκBα, p50, MMP9); none v weak/moderate intensity of nuclear and cytoplasmic staining in the majority of cells (phospho-IκBα); and <10 % v > =10 % of positive cells in nuclei (phospho-p65). (GIF 316 kb)
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