Chk1与Chk2高表达对DADS阻滞人胃癌BGC823细胞G2/M期的影响  

Effect of Chk1/2 Overexpression on the G2/M Arrest in Gastric Cancer BGC823 Cells Induced by Diallyl Disulfide

作　　者：王莉[1,2] 曾颖[2] 夏红 刘芳[1] 苏波[1] 曾希 凌晖[1] 苏琦[1] WANG Li;ZENG Ying;XIA Hong;LIU Fang;SU Bo;ZENG Xi;LING Hui;SU Qi(Cancer Research Institute,University of South China,Hunan Hengyang 421001,China;Department of Oncology,the People′s Hospital of Zouping,Shandong Zouping 256200,China)

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001 [2]邹平县人民医院肿瘤科,山东邹平256200

出　　处：《中国医药导刊》2019年第2期99-103,共5页Chinese Journal of Medicinal Guide

基　　金：国家自然科学基金(项目编号:81641112;项目名称:二烯丙基二硫靶向阻断Rac1信号通路介导的EMT抑制胃癌细胞迁移与侵袭);国家自然科学基金(项目编号:81374013;项目名称:二烯丙基二硫上调RORα拮抗Wnt/β-catenin通路抑制人胃癌细胞EMT与迁移侵袭)

摘　　要：目的:在建立Chk1/2基因高表达人胃癌BGC823细胞的基础上,探讨Chk1/2基因在二烯丙基二硫(DADS)诱导人胃癌BGC823细胞G2/M期阻滞中的作用。方法:采用流式细胞术检测DADS作用于Chk1/2高表达BGC823细胞周期分布情况。Western blot检测Chk1、p-Chk1、Chk2、p-Chk2、Cdc25C与Cyclin B1表达变化。结果:流式细胞术显示,Chk1高表达组G2/M期细胞较对照组与空载体组增加(P<0.05)。而15 mg·L^(-1) DADS处理后,各组G2/M期细胞较处理前增加,Chk1高表达组较空载体组差异有统计学意义(P<0.05)。Chk2高表达组G2/M期细胞较对照组与空载体组差异无统计学意义(P>0.05)。而DADS处理Chk2高表达组后,G2/M期细胞较对照组与空载体组差异有统计学意义(P<0.05)。Western blot显示,Chk1/2蛋白及p-Chk2表达水平不受DADS的影响,但p-Chk1呈时间依赖性上调(P<0.05)。DADS处理Chk1高表达BGC823细胞12、24、36、48 h后,Cdc25C磷酸酶与Cyclin B1表达较对照组呈时间依赖性下降(P<0.05)。结论:DADS阻滞人胃癌BGC823细胞G2/M期与激活Chk1/Cdc25C/Cyclin B1通路有关。Chk1高表达可增强DADS阻滞G2/M期细胞的作用,而Chk2高表达对DADS无影响。Objective : To investigate the role of Chk1/2 on the G2/M phase arrest in gastric cancer BGC823 cells induced by diallyl disulfide (DADS) on the basis of the establishment of Chk1/2 gene overexpression BGC823 cells. Methods : Flow cytometry was used to detect the cell cycle distribution in Chk1/2 overexpression BGC823 cells treated by DADS.The expressions of Chk1, p-Chk1,Chk2, p-Chk2, Cdc25C and Cyclin B1 were detected by Western blot. Results :Flow cytometry showed that the G2/M phase cells in Chk1 overexpression group were increased compared with the control group and the vector group ( P <0.05).After 15 mg·L^-1 DADS treatment,G2/M phase cells in each group were increased compared with those before treatment,and there was statistically significant of the difference between the Chk1 overexpression group and the vector group ( P <0.05). The G2/M phase cells in the Chk2 overexpression group exhibited no statistically significant of the difference with the control group and the vector group ( P >0.05). The G2/M phase cells were different from those in the control group and the vector group after DADS treated Chk2 overexpression group ( P <0.05). Western blot showed that the expression level of Chk1/2 protein and p-Chk2 was not affected by DADS, but the p-Chk1 was time-dependent increased( P <0.05). The expression of Cdc25C and Cyclin B1 showed time-dependent decrease after DADS treated Chk1 overexpression BGC823 cells for 12,24,36 and 48 h ( P <0.05). Conclusion :DADS arrest the G2/M phase in BGC823 cells is related to the activation of Chk1/Cdc25C/Cyclin B1 pathway.Chk1 overexpression can enhance the role of DADS arrest G2/M phase, while Chk2 overexpression has no effect on DADS.

关 键 词：人胃癌BGC823细胞 二烯丙基二硫 Chk1/Chk2高表达 G2/M期 

分 类 号：R735.2[医药卫生—肿瘤]