塞莱昔布对舌鳞癌细胞Cal-27增殖的抑制作用

Inhibitory effect of celecoxib on Cal-27 tongue squamous cell carcinoma cell proliferation

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、100μmol/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(0μmol/L)与实验组(10、20、40μmol/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、40μmol/L)CELE给药作用24 h后,以及经40μmol/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(phos-pho-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,40μmol/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>10μmol/L CELE组>20μmol/L CELE组>40μmol/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。

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 con-centration of CELE,samples were divided into a control group(0μmol/L)and experimental groups(10,20,and 40μmol/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 homo-logue deleted on chromosome ten(PTEN),phospho-protein kinase B(p-AKT)(Thr308),c-Myc,cyclin D1 and other pro-teins in Cal-27 cells after 24 h of treatment with different doses of CELE(10,20,and 40μmol/L)and after 6,12,and 24 h of treatment with 40μmol/L CELE.Results The different concentrations of CELE were able to inhibit the prolif-eration of Cal-27 cells,and the higher the concentration of CELE was,the more significant the inhibition of the prolifera-tion of Cal-27 cells was.The cell survival rates of cells exposed to 40μmol/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 decreas-ing order were the control group,10μmol/L CELE,20μmol/L CELE,and 40μmol/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 expres-sion of PTEN protein increased in the Cal-27 cells after administration of CELE at different concentrations.Conclu-sion 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.