十字孢碱对口腔鳞状细胞癌CAL27细胞增殖和凋亡的影响

Effect of staurosporine on cell proliferation and apoptosis in oral squamous cell carcinoma cell line CAL27

目的探讨十字孢碱(staurosporine,ST)对人口腔鳞状细胞癌细胞株CAL27增殖和凋亡的影响及其机制。方法采用CCK-8法检测不同浓度的ST对CAL27细胞增殖的抑制作用,DAPI荧光染色法观察不同浓度的ST对CAL27细胞凋亡的改变,流式细胞术检测不同浓度ST对CAL27细胞凋亡率和周期分布的影响;蛋白免疫印迹法检测不同浓度ST对CAL27细胞周期蛋白cyclin D1、Cdk4、Cdk6、p21表达的影响。结果 ST对CAL27细胞的增殖有明显抑制作用(P<0.05),抑制作用具有药物浓度依赖性。通过DAPT染色,ST能明显诱导细胞凋亡及凋亡形态改变。CAL27细胞出现Annexin V-FITC/PI流式细胞术观察发现与对照组相比,ST可诱导CAL27细胞凋亡率明显增加(P<0.05),并引起细胞周期阻滞于G2/M期(P<0.05)。蛋白免疫印迹法检测结果显示,经ST处理细胞后,cyclin D1、Cdk4和Cdk6蛋白表达明显降低,p21蛋白表达升高(P<0.05)。结论 ST可显著抑制口腔鳞状细胞癌CAL27细胞增殖并诱导其凋亡,其机制考虑是通过阻滞细胞周期于G2/M期,进而抑制肿瘤细胞生长并促进细胞凋亡。

Objective To study the effect of staurosporine （ST） on cell proliferation and apoptosis in oral squamous cell carcinoma cell line CAL27 and to explore the possible mechanisms. Methods After CAL27 cells were treated with different concentrations of ST for different durations, cell proliferation was assessed by CCK-8 method, cell apoptosis was detected by AnnexinV-FITC Apoptosis Detection Kit and flow cytometry, cell cycle distribution was investigated by flow cytometry, and expression of eyclin B1 and Cdkl proteins was detected by Western blot. Results ST significantly inhibited the proliferation of CAL27 cells （P〈0.05） in a dosedependent manner and the apoptosis rate of CAL27 cells treated with ST increased significantly compared with the controlled cells （P〈0.05）. Treatment with ST induced cell cycle arrest in G2/M phase （P〈0.05）. Moreover, ST treatment significantly decreased the expression of cyelin D1 ,Cdk4 and Cdk6 proteins and increased the expression of p21 in CAL27 cells （P〈0.05）. Conclusion ST significantly inhibits proliferation and induces apoptosis of CAL27 cells possibly by inducing cell cycle arrest in G2/M phase.