脱氧雪腐镰刀菌烯醇对胃癌细胞增殖及细胞周期的影响  

Effects of Deoxynivalenol on Proliferation and Cell Cycle Distribution of Human Gastric Carcinoma Cell Line

作　　者：邢欣[1] 刘静[1] 邢凌霄[1] 刘惠民[1] 周炳娟[1] 王俊灵[1] 严霞[1] 张祥宏[1] 

机构地区：[1]河北医科大学病理研究室,石家庄050017

出　　处：《细胞生物学杂志》2007年第5期729-735,共7页Chinese Journal of Cell Biology

摘　　要：探讨脱氧雪腐镰刀菌烯醇(DON)对体外培养的两株胃癌细胞(SGC-7901和BGC-823)增殖及细胞周期的影响。采用细胞培养、噻唑蓝(MTT)比色法、流式细胞定量检测(FCM)、蛋白质免疫印迹(Western印迹)以及免疫细胞化学染色(ICH)等方法,研究不同浓度DON处理72h对体外培养胃癌细胞的增殖、细胞周期及细胞周期相关蛋白——细胞周期蛋白依赖性激酶抑制蛋白(CKIs)P21WAF/CIP1和细胞周期蛋白E(cyclinE)表达的影响。MTT法检测结果显示DON可明显抑制两株胃癌细胞的增殖,SGC-7901和BGC-823细胞100、500、1000μg/LDON处理组的增殖抑制率分别为4.28%、36.20%、45.35%和14.89%、32.30%、51.61%。FCM检测结果显示,给予1000μg/LDON处理72h可使两株细胞周期阻滞在G2/M期。在100～1000μg/L浓度范围内,两株细胞P21WAF/CIP1表达量均高于对照组,P21WAF/CIP1的表达与DON浓度呈显著正相关关系(SGC-7901细胞:r=0.886,P<0.01;BGC-823细胞:r=0.943,P<0.01);两株细胞的细胞周期蛋白E表达量均低于对照组,与DON浓度有明显剂量依赖关系(SGC-7901细胞:r=-0.923,P<0.01;BGC-823细胞:r=-0.854,P<0.01)。Western印迹及免疫细胞化学检测进一步证实了DON处理对蛋白质表达的影响。综合结果表明,DON可抑制体外培养胃癌细胞的增殖活性,G2/M期阻滞、P21WAF/CIP1表达增高及细胞周期蛋白E表达下降可能是DON抑制胃癌细胞增殖的可能机制,DON对分化程度不同的胃癌细胞的影响没有明显差别。To explore the putative effects of deoxynivalenol （DON） on the proliferation and the cell cycle distribution of SGC-7901 and BGC-823 cells in vitro. SGC-7901 and BGC-823 cells were treated with DON at different concentrations for 72 h. The effects of DON on the cell proliferation and cell cycle distribution of the two cell lines in vitro were determined with MTT and flow cytometric （FCM） DNA analysis, while that on the expression of P21WAF/CIP1 and cyclin E at protein level were studied with immunocytochemical （ICH） staining and Western blotting. MTT results showed that DON treatment could significantly inhibit the proliferation of the two cell lines. The inhibition rate of DON at 100, 500 and 1 000 μg/L was 4.28%, 36.20% and 45.35% for SGC-7901 and 14.89%, 32.30% and 51.61% for BGC-823 respectively. FCM cell cycle analysis revealed that G2/M arrest could be induced by 1 000 lag/L DON treatment for 72 h for both cell lines. Within the concentration range from 100μg/L to 1 000μg/ L, DON could significantly increase the expression of P21WAF/CIP1. A significant dose-effect correlation could be found between DON concentration and the intensity of P21WAF/CIP1 expression at protein level for both cell lines （SGC-7901： r=0.886, P〈0.01; BGC-823： r=0.943, P〈0.01）. While the expression of cyclin E was significantly decreased by DON in a dose-dependent way in the two cell lines （SGC-7901： r=-0.923, P〈0.01; BGC-823： r= -0.854, P〈0.01）. The results of immunocytochemical staining and Western blotting confirmed that DON could increase the expression of P21WAF/CIP1, while decrease that of cyclin E in both two cell lines. Thus, the results in this study suggested that DON could significantly inhibit the proliferation of gastric carcinoma SGC-7901 and BGC-823 cells in vitro, and the possible mechanism of the proliferation inhibiting effects of DON may be G2/M arrest and the increase of the expression of P21wAF/CIP1 and decrease of cyclin E. No significant differences were found in the effects

关 键 词：脱氧雪腐镰刀菌烯醇 细胞周期 P21^WAF/CIP1. 细胞周期蛋白E 

分 类 号：R735.2[医药卫生—肿瘤]