沉默Chk1基因对DADS阻滞人胃癌BGC823细胞G2/M期的影响

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沉默Chk1基因对DADS阻滞人胃癌BGC823细胞G2/M期的影响

谭亚丽,夏红#,曾颖,刘芳,苏波,凌晖,苏琦

南华大学肿瘤研究所湖南省中医高等专科学校附属一医院肿瘤科,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:2018-06-13 修回日期:2018-06-13 出版日期:2018-06-25
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 the G2/M Arrest Induced by Diallyl Disulfide in Gastric Cancer BGC823 Cells of Silencing Chk1 Gene

TAN YaLi,XIA Hong#,ZENG Ying,LIU Fang,SU Bo,LING Hui and SU Qi

Cancer Research Institute, University of South China Department of Oncology, First Affiliated Hospital, Hunan college of traditional Chinese medicine,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:2018-06-13 Revised:2018-06-13 Online:2018-06-25

摘要/Abstract

摘要： 目的：探讨沉默Chk1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RNAi 技术沉默BGC823细胞Chk1基因，q-PCR与Western blot检测Chk1 mRNA与蛋白表达。流式细胞术检测DADS作用与Chk1基因沉默BGC823细胞周期分布情况。Western blot检测Cdc25C与cyclinB1表达。结果：流式细胞术显示，15 mg·L-1 DADS作用BGC823细胞12、24、36、48 h后，G2/M期细胞呈时间依赖性增加(P<0.05)。Chk1沉默BGC823细胞Chk1 mRNA与蛋白表达下调(P<0.05)。沉默Chk1后，G2/M期细胞较对照组降低(P<0.05)。DADS处理后，对照组G2/M细胞高于Chk1沉默组(P<0.05)。Chk1基因沉默后Cdc25C和CyclinB1表达较对照组增加(P<0.05)。DADS处理对照组后，Cdc25C和CyclinB1表达较未处理组降低(P<0.05)。DADS处理Chk1沉默组Cdc25C和CyclinB1表达较Chk1沉默下调(P<0.05)。结论：Chk1沉默可通过促进Cdc25C和CyclinB1表达减弱DADS阻滞G2/M的作用，DADS阻滞G2/M的检查点是Chk1。

Abstract: Objective： To investigate the effect of silent Chk1 gene on the G2/M block of gastric cancer BGC823 cells induced by diallyl disulfide (DADS). Methods： Using RNAi technology to silence the Chk1 gene， q-PCR and Western blot were used to detect Chk1 mRNA and protein expression. Flow cytometry was used to detect the cell cycle distribution of DADS and silencing Chk1 BGC823 cells. The expression of Cdc25C and cyclinB1 were detected by Western blot. Results： Flow cytometry showed that the cells of G2/M increased in time dependent after the DADS effected BGC823 cells for 12， 24， 36 and 48 h (P<0.05). Chk1 mRNA and protein expression were down-regulated (P<0.05). After silencing Chk1， G2/M cells were lower than that of the control group (P<0.05). After DADS treatment， G2/M cells in the control group were higher than the Chk1 silent group (P<0.05). After silencing Chk1， the expression of Cdc25C and CyclinB1 was increased compared with the control group (P<0.05). After DADS treatment， the expression of Cdc25C and CyclinB1 was lower than that in the untreated group (P<0.05). The expression of Cdc25C and CyclinB1 in DADS treatment Chk1 silent group was significantly lower than that of Chk1 silent group (P<0.05). Conclusion： Chk1 silencing through promoting the expression of Cdc25C and CyclinB1 can weaken the G2/M arrest induced by DADS， and the detection point of DADS block G2/M is Chk1.

谭亚丽,夏红#,曾颖,刘芳,苏波,凌晖,苏琦. 沉默Chk1基因对DADS阻滞人胃癌BGC823细胞G2/M期的影响[J]. .

TAN YaLi,XIA Hong#,ZENG Ying,LIU Fang,SU Bo,LING Hui and SU Qi. Effect of the G2/M Arrest Induced by Diallyl Disulfide in Gastric Cancer BGC823 Cells of Silencing Chk1 Gene[J]. .

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