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

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

王莉,曾颖,夏红,刘芳,苏波,曾希,凌晖,苏琦

南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所

收稿日期:2019-02-12 修回日期:2019-03-02 出版日期:2019-02-25
基金资助:
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）

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

WANG Li,ZENG Ying,XIA Hong,LIU Fan,SU Bo,ZENG Xi,LING Hui and SU Qi

Cancer Research Institute, University of South China, Department of Oncology, the People’s Hospital of Zouping,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China

Received:2019-02-12 Revised:2019-03-02 Online:2019-02-25

摘要/Abstract

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。

Abstract: 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.

王莉,曾颖,夏红,刘芳,苏波,曾希,凌晖,苏琦. Chk1与Chk2高表达对DADS阻滞人胃癌 BGC823细胞G2/M期的影响[J]. .

WANG Li,ZENG Ying,XIA Hong,LIU Fan,SU Bo,ZENG Xi,LING Hui and SU Qi. Effect of Chk1/2 Overexpression on the G2/M Arrest in Gastric Cancer BGC823 Cells Induced by Diallyl Disulfide[J]. .

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