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目的探讨脂多糖(LPS)对喉癌细胞高迁移率族蛋白B1(HMGB1)释放的诱导作用。方法采用酶联免疫吸附试验、实时荧光定量PCR和Western blot检测LPS诱导后人喉癌细胞株Hep-2培养上清液中HMGB1含量、细胞HMGB1mRNA表达水平和细胞Toll样受体4(TLR4)、核因子-κB(NF-κB)蛋白表达水平的变化,观察TLR4单克隆抗体、NF-κB抑制剂二硫代氨基甲酸吡咯烷(PDTC)对HMGB1释放的影响。结果 LPS诱导12h和18h后,HMGB1mRNA表达水平和HMGB1含量均升高,且呈时间依赖性;LPS诱导24h后,TLR4及NF-κB p65蛋白明显升高。TLR4单克隆抗体和PDTC对LPS诱导的HMGB1释放有不完全的抑制作用。结论 LPS诱导喉癌细胞释放HMGB1;其机制可能与TLR4/NF-κB信号通路有关。
Objective To investigate the effect of lipopolysaccharide (LPS) on the release of high mobility group box 1 (HMGB1) from laryngeal cancer cells. Methods Enzyme-linked immunosorbent assay, real-time fluorescence quantitative PCR and Western blot were used to detect the HMGB1 content, the cell HMGB1 mRNA expression level and the cell-toll-like receptor 4 (TLR4) in human laryngeal carcinoma cell line Hep-2. The expression of nuclear factor-κB (NF-κB) protein was observed. The effects of TLR4 monoclonal antibody and NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) on the release of HMGB1 were observed. Results After LPS induction for 12 h and 18 h, HMGB1 mRNA and HMGB1 levels were increased in a time-dependent manner. After 24 h of LPS induction, TLR4 and NF-κB p65 protein were significantly increased. TLR4 monoclonal antibody and PDTC have incomplete inhibitory effect on LPS-induced HMGB1 release. Conclusion LPS induces the release of HMGB1 from laryngeal carcinoma cells; its mechanism may be related to TLR4/NF-κB signaling pathway.