DON对胃癌细胞HGC-27细胞周期和凋亡的影响

Effects of Deoxynivalenol on Cell Cycle and Apoptosis of Human Gastric Carcinoma Cell Line HGC-27 in vitro

目的探讨脱氧雪腐镰刀菌烯醇（DON）对体外培养胃癌细胞HGC-27细胞周期和凋亡的影响。方法体外培养的胃癌细胞HGC-27经50、100、1000、2000μg／L DON处理12h、24h、48h，应用流式细胞术（FCM）进行细胞周期分析，检测凋亡情况及其量效关系，Western blot检测凋亡相关蛋白Bax、Bcl-2和Caspase-3的表达情况。结果FCM检测结果表明，较高浓度（1000和2000μg／L）DON处理对细胞周期分布的影响随处理时间不同有明显差异。DON处理12h，可明显降低G0／G1细胞百分率、增加S期细胞百分率。处理时间延长至24h和48h，则表现为G0／G1细胞百分率增加，S期细胞百分率降低（P〈0．05）。DON处理24h和48h，各DON实验组HGC-27细胞凋亡率均高于对照组，在50～2000ug／L浓度范围内，凋亡率随着DON处理浓度的升高而升高。Western blot检测凋亡相关蛋白结果显示，DoN处理12、24和48h，Bax和Caspase-3蛋白表达均明显升高，而Bcl-2蛋白表达降低。结论DON处理可影响体外培养的胃癌细胞象HGC-27细胞的细胞周期分布，其作用依DON浓度和作用时间的不同而有差异。同时DON可诱导HGC-27细胞凋亡，上调Bax和Caspase-3蛋白表达和下调Bcl-2蛋白表达可能是其诱导细胞凋亡的机制之一。

Objective To explore the putative effect of deoxynivalenol on cell cycle and apoptosis of human gastric carcinoma cell line HGC-27 in vitro. Methods HGC-27 cells were treated with DON at different concentrations（50,100,l 000,2 000μg/L） for 12, 24 and 48 hours, and then cells were harvested for analysis of apoptosis, cell cycle distribution and expression of Bax, Bcl-2 and Caspase-3 with flow cytometric （FCM） and Western Blot methods. Results At relatively higher concentrations （1000 and 2 000 g/L）, DON could affect cell cycle distribution in a time dependent manner. DON treatment for 12 h, the percentages of cells in G0/G1 phases was significantly decreased and that in S phase increased. As the treatment time lasted for 24 h and 48 h, the percentages of cells in G0/G1 phases were significantly increased while that in S phase decreased （P〈0.05）. No significant effects on G2/M phase could be found. The apoptosis rates in all DON treated groups for 12 h, 24 h and 48 h were all higher than that in control. Though statistical significance in the difference of apoptosis rate between control and the DON treated groups was not found 12 h after DON treatment but it could be seen 24 and 48 h after DON treatment （P〈0. 05）. Significant dose-effect relationships were found between DON concentrations and the apoptosis rates. Western Blot analysis showed bax and caspase-3 expression was up-regulated and bcl-2 expression was down-regulated in DON treated HGC-27 cells in vitro. Conclusion DON could significantly affect the cell cycle distribution of HGC-27 cells in vitro and the effects of DON on cell cycle varied as the concentratrion and treatment time of DON changed. DON treatment could induce apoptosis of HGC-27 cell possibly by up-regulating bax and caspase-3 expression and down-regulating bcl-2 expression.