奥沙利铂对人舌鳞癌CAL27细胞增殖和凋亡的影响及其机制

doi:10.13481/j.1671-587x.20200205

摘要/Abstract

摘要： 目的：探讨奥沙利铂（Oxaliplatin）对人舌鳞状细胞癌（简称舌鳞癌）CAL27细胞增殖和凋亡的影响，并阐明其潜在作用机制。方法：采用不同浓度（0、10、20、40和80 mg·L^-1）Oxaliplatin处理CAL27细胞，分别作为对照组和10、20、40及80 mg·L^-1 Oxaliplatin组，采用结晶紫染色法观察各组CAL27细胞形态表现，CCK-8法检测各组CAL27细胞增殖率，流式细胞术检测不同细胞周期CAL27细胞百分比和凋亡率，实时荧光定量PCR法检测各组CAL27细胞中周期蛋白依赖性激酶2（CDK2）和细胞周期蛋白E（cyclin E）mRNA表达水平，Western blotting法检测各组CAL27细胞中抗凋亡蛋白survivin和促凋亡蛋白Bax的表达水平。结果：与对照组比较，40 mg·L^-1 Oxaliplatin组CAL27细胞出现一定程度的皱缩，80 mg·L^-1 Oxaliplatin组CAL27细胞明显失去原有形态，细胞间连接丧失。与对照组比较，10、20、40和80 mg·L^-1 Oxaliplatin组CAL27细胞增殖率明显降低（P<0.05或P<0.01）。与对照组比较，40 mg·L^-1 Oxaliplatin组G₀/G₁期CAL27细胞百分比明显升高（P<0.01），CDK2和cyclin E mRNA表达水平明显降低（P<0.05）。与对照组比较，40与80 mg·L^-1 Oxaliplatin组CAL27细胞凋亡率明显升高（P<0.01）。与对照组比较，40和80 mg·L^-1 Oxaliplatin组CAL27细胞中survivin蛋白表达水平明显降低（P<0.01），Bax蛋白表达水平则明显升高（P<0.05或P<0.01）。结论：Oxaliplatin能够抑制CAL27细胞增殖，促进细胞凋亡，其机制可能与细胞发生G₀/G₁期阻滞以及survivin蛋白表达水平降低和Bax蛋白表达水平升高有关。

关键词: 舌鳞状细胞癌, 奥沙利铂, 细胞增殖, 细胞凋亡, 细胞周期

Abstract: Objective: To explore the effect of Oxaliplatin on the proliferation and apoptosis of human tongue squamous cell carcinoma CAL27 cells, and to clarify its underlying mechanism. Methods: The CAL27 cells were treated with different concentrations of Oxaliplatin (0,10, 20, 40 and 80 mg·L^-1) and used as control group and 10, 20, 40 and 80 mg·L^-1 Oxaliplatin groups. The morphology of CAL27 cells in various groups were observed by crystal violet staining. The proliferation rates of the CAL27 cells in various groups were detected by CCK-8 assay. The percentages of CAL27 cells in different cell cycles and the apoptotic rates in various groups were detected by flow cytometry. The expression levels of cyclin-dependent kinase 2(CDK2)and E-type cyclin(cyclin E) mRNA in the CAL27 cells in various groups were analyzed by Real-time fluorescence quantitative PCR. The expression levels of anti-apoptotic protein survivin and pro-apoptotic protein Bax in the CAL27 cells in various groups were determined by Western blotting method. Results: Compared with control group, the CAL27 cells in 40 mg·L^-1 Oxaliplatin group wrinkled; the cells in 80 mg·L^-1 Oxaliplatin group lost the former morphology, and the intercellular connections of cells disapeared. Compared with control group, the proliferation rates of CAL27 cells in 10, 20, 40 and 80 mg·L^-1 Oxaliplatin groups were significantiy decreased (P<0.05 or P<0.01).Compared with control group, the percentage of CAL27 cells in G₀/G₁ phase in 40 mg·L^-1 Oxaliplatin group was significantly increased (P<0.01), and the expression levels of CDK2 and cyclin E mRNA were significantly decreased (P<0.05).Compared with control group, the apoptptic rates of CAL27 cells in 40 and 80 mg·L^-1 Oxaliplatin groups were markedly increased (P<0.01). Compared with control group, the expression levels of survivin protein in CAL27 cells in 40 and 80 mg·L^-1 Oxaliplatin groups were significantly decreased (P<0.01), and the expression levels of Bax protein were obviously increased (P<0.05 or P<0.01). Conclusion: Oxaliplatin can inhibit the proliferation and induce the apoptosis of CAL27 cells, and its mechanism may be related to arresting G₀/G₁ phase, up-regulating the expression level of survivin and down-regulating the expression level of Bax.

Key words: tongue squamous cell carcinoma, Oxaliplatin, cell proliferation, apoptosis, cell cycle

中图分类号:

R739.86

张楚, 郝苗, 王会俞, 王晓峰, 张天夫. 奥沙利铂对人舌鳞癌CAL27细胞增殖和凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2020, 46(02): 233-239.

ZHANG Chu, HAO Miao, WANG Huiyu, WANG Xiaofeng, ZHANG Tianfu. Effect of Oxaliplatin on proliferation and apoptosis of human tongue squamous cell carcinoma CAL27 cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 233-239.

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