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.
Department of Laboratory Medicine, the First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China.
Xiaojie
X
Zhang
Juan
J
Xu
Xing
X
Ke
Shuping
S
Zhang
Peijun
P
Huang
Ting
T
Xu
Lei
L
Huang
Jianfang
J
Lou
Xinhui
X
Shi
Ruihong
R
Sun
Fang
F
Wang
Shiyang
S
Pan
D000328:Adult; D016022:Case-Control Studies; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D007249:Inflammation / Q000097:blood / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008875:Middle Aged; D053594:Myeloid Differentiation Factor 88 / Q000097:blood; D010051:Ovarian Neoplasms / Q000097:blood* / Q000276:immunology; D051195:Toll-Like Receptor 2 / Q000096:biosynthesis / Q000097:blood; D051216:Toll-Like Receptor 6 / Q000096:biosynthesis / Q000097:blood; D051193:Toll-Like Receptors / Q000096:biosynthesis / Q000097:blood*
C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; C495342:TLR2 protein, human; C495368:TLR6 protein, human; D051195:Toll-Like Receptor 2; D051216:Toll-Like Receptor 6; D051193:Toll-Like Receptors