摘要
目的:探讨十字孢碱(staurosporine,ST)对人的胰腺癌细胞株Panc-1增殖和凋亡的影响及其机制.方法:采用CCK-8法检测ST对Panc-1细胞的增殖抑制作用;Hoechst 33258荧光染色观察ST对Panc-1细胞凋亡形态的影响;流式细胞术检测不同浓度ST对Panc-1细胞凋亡率和周期分布的影响;蛋白质印迹法检测不同浓度ST对Panc-1细胞周期蛋白cyclin A、cyclin D1、Cdk4和P21表达的影响.结果:ST对Panc-1细胞的增殖有明显抑制作用(P<0.05),并且这种抑制作用呈浓度和时间依赖性.Hoechst 33258荧光染色观察发现,ST能明显诱导Panc-1细胞出现典型的凋亡形态学改变.流式细胞术检测发现,与对照组相比,ST可诱导Panc-1细胞凋亡率明显增加(P<0.05),并引起细胞周期阻滞于G1期(P<0.05).蛋白质印迹法检测结果显示,Panc-1细胞经ST处理后cyclin D1、Cdk4蛋白表达明显降低(P<0.05),P21蛋白表达则明显增加(P<0.05);低浓度ST上调Panc-1细胞中cyclin A的表达,而高浓度ST则下调cyclin A的表达(P<0.05).结论:ST可显著抑制胰腺癌Panc-1细胞增殖并诱导其凋亡,其机制可能是通过阻滞细胞周期于G1期,进而抑制肿瘤细胞生长并促进细胞凋亡.
AIM： To investigate the effect of staurosporine （ST） on cell proliferation and apoptosis in pancreatic cancer cell line Panc-1, and to explore the possible mechanisms involved.
METHODS： After Panc-1 cells were treated with different concentrations of ST for different durations, cell proliferation was assessed by CCK-8 method, cell apoptosis was detected by Hoechst 33258 staining and flow cytometry, cell cycle distribution was investigated by flow cytometry, and expression of cyclin A, cyclin D1, Cdk4 and P21 proteins was detected by Western blot.
RESULTS： ST significantly inhibited the proliferation of Panc-1 cells （P 〈 0.05） in a concentration- and time-dependent manner. Treatment with ST significantly induced apoptosis of Panc-1 cells, and the apoptosis rate of Panc-1 cells treated with ST increased significantly compared with control cells （P 〈 0.05）. Treatment with ST induced cell cycle arrest in G1 phase （P 〈 0.05）. Moreover, ST treatment significantly decreased the expression of cyclin D1 and Cdk4 proteins and increased the expression of P21 protein in Panc-1 cells （all P 〈 0.05）. Low concentrations of ST raised the expression of cyclin A in Panc-1 cells, while high concentrations of ST reduced the expression of cyclin A （P 〈 0.05）.
CONCLUSION： ST significantly inhibits proliferation and induces apoptosis of Panc-1 cells possibly by inducing cell cycle arrest in G1 phase.
出处
《世界华人消化杂志》
CAS
北大核心
2013年第25期2578-2584,共7页
World Chinese Journal of Digestology
基金
国家自然科学基金面上资助项目
Nos.81071775
81172064
81160311
81001068
81101621
湖北省自然科学重点基金资助项目
No.2011CDA030
中国博士后科学基金面上资助项目
No.2013M531983~~
