Contribution of FPR and TLR9 to hypoxia-induced chemoresistance of ovarian cancer cells
Contribution of FPR and TLR9 to hypoxia-induced chemoresistance of ovarian cancer cells
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DOI：
10.2147/ott.s190118
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发表时间：
2019-01-01
影响因子：
4
通讯作者：
Chen, Jianhong
中科院分区：
文献类型：
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作者：
Cai, Yongqing;Huang, Jian;Chen, Jianhong
背景/目的：本研究旨在探讨甲酰基肽受体（FPR）和toll样受体9 （TLR9）在缺氧诱导的卵巢癌细胞化疗耐药中的作用及其机制。材料与方法：将SKOV3细胞缺氧处理24h，取上清液刺激常氧培养的SKOV3细胞，CCK8检测细胞生长抑制率。应用TLR9 CpG ODN激动剂和FPR fMLF激动剂研究SKOV3细胞对顺铂的化学敏感性。同时用FPR拮抗剂t-Boc或TLR9拮抗剂CQ处理这些细胞。Western blot检测FPR、TLR9、MRP、P-gp、p53、Beclin-1蛋白水平。采用免疫荧光染色法观察TLR9在SKOV3细胞中的分布。结果：低氧暴露降低了顺铂对SKOV3细胞的抑制率。WB结果显示，FPR和TLR9在人卵巢癌组织和SKOV3细胞中均有表达，且随缺氧时间的延长而升高。用fMLF或ODN2006刺激SKOV3后，顺铂诱导的抑制率明显降低。tBoc和CQ可显著减弱缺氧上清诱导的SKOV3细胞耐药。低氧上清液显著增加SKOV3细胞的MRP、P-gp、p53和Beclin-1蛋白，tBoc显著降低这些蛋白。结论：缺氧可上调人卵巢癌细胞系FPR和TLR9的表达，促进两种受体配体的释放。FPR和TLR9可能是卵巢癌细胞化疗致敏的新靶点。
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 cancer cells.