Abstract:

Objective To explore the inhibitory effect of celecoxib (CELE) on the proliferation of tongue squamous cell carcinoma Cal-27 cells and its mechanism. Methods A CCK-8 assay was used to investigate the cytotoxicity of different concentrations CELE(10, 20, 40, 60, 80, and 100 mol/L) at 24 and 48 h in Cal-27 cells. According to the concentration of CELE, samples were divided into a control group (0 μmol/L) and experimental groups (10, 20, and 40 μmol/L), and cell invasiveness was detected by the Transwell method. The expression levels of c-Myc and Cyclin D1 mRNA were detected with qPCR, and western blots were used to detect the expression of phosphate and tension homologue deleted on chromosome ten (PTEN), phospho-protein kinase B (p-AKT) (Thr308), c-Myc, cyclin D1 and other proteins in Cal-27 cells after 24 h of treatment with different doses of CELE (10, 20, and 40 μ mol/L) and after 6, 12, and 24 h of treatment with 40 μmol/L CELE. Results The different concentrations of CELE were able to inhibit the proliferation of Cal-27 cells, and the higher the concentration of CELE was, the more significant the inhibition of the proliferation of Cal-27 cells was. The cell survival rates of cells exposed to 40 μmol/L CELE were 80% and 75% after 24 and 48 h, respectively. In the four groups of patients, the number of invasive cells was compared, and the results in decreasing order were the control group, 10 μmol/L CELE, 20 μmol/L CELE, and 40 μmol/L CELE. The expression level of c-Myc, cyclin D1 mRNA and the protein in P-AKT (Thr308), c-Myc, and cyclin D1 significantly decreased and the expression of PTEN protein increased in the Cal-27 cells after administration of CELE at different concentrations. Conclusion CELE can inhibit the proliferation of Cal-27 cells, possibly through inhibition of the expression of proliferation signal factors, such as c-Myc and cyclin D1, by activating the PTEN signaling pathway.

Key words: celecoxib, tongue squamous cell carcinoma, tongue squamous cell carcinoma Cal-27, phosphate and tension homology deleted on chromsome ten, protein kinase B, c-Myc, Cyclin D1, proliferation, invasive

摘要：

目的 探讨塞莱昔布（celecoxib,CELE）对舌鳞癌细胞Cal-27增殖的抑制作用及其机制。方法 CCK-8检测不同浓度（10、20、40、60、80、100 μmol/L）CELE作用24 h、48 h后对舌鳞癌细胞Cal-27的细胞毒性,依据CELE的浓度,分为对照组（0 μmol/L）与实验组（10、20、40 μmol/L）。采用Transwell检测细胞侵袭性;荧光定量PCR（qPCR）检测c-Myc、细胞周期蛋白（Cyclin D1）的mRNA的表达水平;Western blot法检测经不同剂量（10、20、40 μmol/L）CELE给药作用24 h后,以及经40 μmol/L的CELE给药作用6、12、24 h后Cal-27细胞的蛋白酪氨酸磷酸酶（phosphate and tension homology deleted on chromsome ten,PTEN）、磷酸化蛋白激酶B（phospho-protein kinase B,p-AKT）（Thr308）、原癌基因蛋白c-Myc、G1/S-特异性周期蛋白-D1（Cyclin D1）等蛋白的表达。结果 不同浓度的CELE对舌鳞癌细胞Cal-27的增殖均具有抑制作用,CELE浓度越高对Cal-27增殖的抑制作用越显著,40 μmol/L CELE作用24、48 h后细胞存活率分别为80%、75%。4组侵袭细胞数比较,对照组>10 μmol/L CELE组>20 μmol/L CELE组>40 μmol/L CELE组;不同浓度的CELE给药作用后,舌鳞癌细胞Cal-27中c-Myc、Cyclin D1 mRNA的表达水平显著降低,p-AKT（Thr308）、c-Myc、Cyclin D1等蛋白表达降低,PTEN蛋白表达升高。结论 CELE对舌鳞癌细胞Cal-27增殖具有抑制作用,其作用机制可能是通过激活PTEN信号通路抑制c-Myc、Cyclin D1等增殖信号因子的表达。

关键词: 塞莱昔布, 舌鳞癌, 舌鳞癌细胞Cal-27, 蛋白酪氨酸磷酸酶, 蛋白激酶B, c-Myc, 细胞周期蛋白1, 增殖, 侵袭