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目的观察选择性环氧化酶-2(COX-2)抑制剂塞来昔布对人舌鳞癌Tca8113细胞的生长抑制、COX-2表达调节及诱导凋亡的作用。方法在Tca8113细胞中分别加入含有不同浓度塞来昔布的培养液,培养24、48、72h后,采用MTT方法测定细胞生长抑制率,采用免疫组化方法观察COX-2蛋白在Tca8113细胞中的表达,应用荧光显微镜观察凋亡细胞的形态学特征,应用Annexin V-FITC/PI双标记法检测细胞的早期凋亡率,相对定量荧光定量PCR检测Tca8113细胞中COX-2 mRNA的表达。结果COX-2蛋白在Tca8113细胞中呈强阳性表达,塞来昔布在抑制细胞生长的同时,抑制Tca8113细胞COX-2蛋白的表达;Tca8113细胞经塞来昔布处理后凋亡细胞多见,与对照组相比,塞来昔布处理组早期凋亡细胞增多有统计学意义;塞来昔布调节COX-2 mRNA表达的作用与对照组相比无统计学意义。结论塞来昔布主要以剂量依赖的方式抑制Tca8113细胞生长,诱导细胞凋亡,其作用与抑制COX-2蛋白表达有关,而对COX-2基因作用较弱,其作用机制有待进一步研究。
Objective To observe the effect of celecoxib, a selective inhibitor of cyclooxygenase-2 (COX-2), on the growth of Tca8113 human tongue squamous cell carcinoma, the regulation of COX-2 expression and the induction of apoptosis. Methods Tca8113 cells were added with different concentrations of celecoxib in culture medium, cultured 24,48,72 h after the MTT method was used to measure the rate of cell growth inhibition by immunohistochemistry COX-2 protein in Tca8113 cells The morphological characteristics of apoptotic cells were observed by fluorescence microscopy. The early apoptosis rate was detected by Annexin V-FITC / PI double labeling method. The expression of COX-2 mRNA in Tca8113 cells was detected by quantitative real-time PCR. Results COX-2 protein was strongly expressed in Tca8113 cells. Celecoxib inhibited the cell growth and inhibited the expression of COX-2 protein in Tca8113 cells. Tca8113 cells were more frequently treated with celecoxib, Compared with the control group, celecoxib treatment group early apoptotic cells increased statistically significant; celecoxib regulation of COX-2 mRNA expression compared with the control group was not statistically significant. Conclusion Celecoxib inhibits the growth and induces apoptosis of Tca8113 cells in a dose-dependent manner. The effect of celecoxib on the expression of COX-2 protein is weak, and the mechanism of action is still to be further studied.