Contribution of FPR and TLR9 to hypoxia-induced chemoresistance of ovarian cancer cells

Yongqing Cai,1 Jian Huang,2 Haiyan Xing,1 Bin Li,1 Ling Li,1 Xianfeng Wang,1 Dan Peng,1 Jianhong Chen1 1Department of Pharmacy, Daping Hospital and Research Institute of Surgery, Army Medical University, Chongqing 400042, China; 2Department of High Altitude Biology and Pathology, High Altitude Military Medical College, Army Medical University, Chongqing 400042, China Background/purpose: The aim of this study was to investigate the role and mechanisms of the formyl peptide receptor (FPR) and the toll-like receptor 9 (TLR9) in hypoxia-induced chemoresistance of human ovarian cancer cells. Materials and methods: SKOV3 cells were exposed to hypoxia for 24 hours, the supernatant was collected to stimulate normoxia-cultured SKOV3, and the inhibition rate of cell growth was detected with CCK8 test. The agonist of TLR9 CpG ODN and the agonist of FPR fMLF were applied to investigate the chemosensitivity of SKOV3 cells to cisplatin. The cells were also treated with FPR antagonist t-Boc or TLR9 antagonist CQ. Western blot was applied to detect protein levels of FPR, TLR9, MRP, P-gp, p53 and Beclin-1. Immunofluorescence staining was applied to observe the distribution of TLR9 in SKOV3 cells. Results: Hypoxia exposure reduced the inhibition rate of cisplatin on SKOV3 cells. WB showed that FPR and TLR9 were expressed in human ovarian cancer tissues and SKOV3 cells, and the levels were increased with longer hypoxia time. After SKOV3 was stimulated with fMLF or ODN2006, cisplatin-induced inhibition rate was significantly decreased. tBoc and CQ significantly attenuated hypoxia supernatant-induced chemoresistance of SKOV3 cells. Hypoxia supernatants significantly increased MRP, P-gp, p53 and Beclin-1 proteins in SKOV3 cells, which were significantly reduced by tBoc. Conclusion: Hypoxia upregulates the expression of FPR and TLR9, and promotes the release of ligands for both receptors in human ovarian cancer cell line. FPR and TLR9 may be noval targets for chemosensitizing to ovarian c