十字孢碱对口腔鳞状细胞癌CAL27细胞增殖和凋亡的影响

›› 2017, Vol. 37 ›› Issue (7): 598-602.

十字孢碱对口腔鳞状细胞癌CAL27细胞增殖和凋亡的影响

孙扬¹,周倩蓉¹,程勇¹,吴启超²,丁小军¹,余优成³

1. 复旦大学附属中山医院
2. 复旦大学附属肿瘤医院
3. 复旦大学附属中山医院口腔科

收稿日期:2016-03-18 修回日期:2016-07-21 出版日期:2017-07-28 发布日期:2017-07-18
通讯作者: 孙扬 E-mail:sunyang_hi@126.com

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

Received:2016-03-18 Revised:2016-07-21 Online:2017-07-28 Published:2017-07-18
Contact: Yang SUN E-mail:sunyang_hi@126.com

摘要/Abstract

摘要： 摘要目的探讨十字孢碱(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。

关键词: 口腔鳞状细胞癌, 十字孢碱, 细胞周期, 细胞凋亡, CAL27细胞

Abstract: Abstract: 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 cyclin B1 and Cdk1 proteins was detected by Western blot. Results ST significantly inhibited the proliferation of CAL27 cells (P < 0.05) in a dose- dependent 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 cyclin 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.

Key words: oral squamous cell carcinoma, Staurosporine, cell cycle, apoptosis, CAL27 cells

中图分类号:

R739.85

孙扬周倩蓉程勇吴启超丁小军余优成. 十字孢碱对口腔鳞状细胞癌CAL27细胞增殖和凋亡的影响[J]. , 2017, 37(7): 598-602.

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