中文摘要：

目的：观察缺氧对喉癌细胞高迁移率族蛋白B1（HMGB1）释放的诱导作用，并探讨其可能的机制。方法：采用人喉癌细胞株Hep-2，观察缺氧（1％O2）培养不同时间对细胞培养上清液中HM（}B1含量、细胞HMGB1蛋白和mRNA的影响，及不同浓度的MAPK信号通路抑制剂（PD98059、SP600125、SB202190）、NF—KB抑制剂（PDTC）对缺氧诱导的HMGB1释放的影响。上清液HMGB1含量和细胞HMGB1蛋白表达水平分别采用酶联免疫吸附试验和Westernblot检测；细胞HMGB1mRNA表达水平采用实时荧光定量PCR法检测。结果：培养上清液中HMGB1含量和细胞HMGB1蛋白表达水平均于缺氧诱导12h后升高，呈时间依赖性；细胞HMGB1mRNA表达水平于缺氧诱导6h开始升高，并随诱导时间延长而进-步升高；20μmol／L PD98059、SP600125和50mg／L PDTC对缺氧诱导的HMGB1释放有不完全的抑制作用。结论：缺氧诱导喉癌细胞释放HMGB1，其机制可能与MAPK／NF-κB信号通路有关。

英文摘要：

Objective：To investigate the extracellular release of high mobility group box 1（HMGB1） in laryn- geal Hep 2 carcinoma cells induced by hypoxia and its possible mechanism. Method： The changes of HMGB1 con centration in the culture medium as well as HMGB1 protein and mRNA expression in Hep-2 cells were investigated after the cells were cultured with 1% O2 for different durations. Inhibitory effects of MAPK pathway inhibitors （PD98059, SP600125, and SB202190） and nuclear NF-κB pathway inhibitor（PDTC） with various concentrations on extracellular HMGB1 release were observed in hypoxia-induced Hep-2 cells. The HMGB1 concentration and HMGB1 protein expression were measured by enzyme linked immunosorbent assay（ELISA） and western blot, re spectively. The H MGB1 mRNA expression was determined by real-time quantitative PCR（RT-PCR）. Result：The HMGB1 concentration in the culture medium and the HMGB1 protein expression in Hep 2 cells increased after the cells were subjected to hypoxia culture for 12 h in a time-dependent manner. The level of HMGB1 mRNA expres- sion in Hep-2 cells increased after the cells were induced by hypoxia for 6h PD98059 and SP600125 with 20 μmol/ L and PDTC with 50 mg/L partly inhibited extracellular release of HMGB1 in hypoxia-cultured Hep-2 cells. Con- clusion： Hypoxia induces laryngeal carcinoma cells to release HMGB1, which may be related to MAPK/NF-κB sig- naling pathway.