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G 0/G 1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G 0/G 1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。
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 G 0/G 1 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 G 0/G 1 phase,up-regulating the expression level of survivin and down-regulating the expression level of Bax.
作者
张楚
郝苗
王会俞
王晓峰
张天夫
ZHANG Chu;HAO Miao;WANG Huiyu;WANG Xiaofeng;ZHANG Tianfu(Department of Stomatology,China-Japan Union Hospital,Jilin University,Changchun 130033,China;Scientific Research Center,China-Japan Union Hospital,Jilin University,Changchun 130033,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2020年第2期233-239,共7页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金资助课题(81903881)
吉林省发改委科研基金资助课题(2018C007)
吉林省卫健委科研基金资助课题(2017F007,2019SCZ006)
吉林大学白求恩计划科研基金资助课题(2018B02)。
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
tongue squamous cell carcinoma
Oxaliplatin
cell proliferation
apoptosis
cell cycle
