沉默Chk2基因对DADS阻滞人胃癌BGC823细胞G2/M期的影响

细胞周期检测点激酶1/2(Chk1/2)在参与G2/M期起着重要作用,本研究探讨沉默Chk2基因对二烯丙基二硫(DADS)诱导人胃癌BGC823细胞G2/M期阻滞的影响。采用RNAi 技术沉默BGC823细胞Chk2基因,q-PCR与Western blot检测Chk2 mRNA与蛋白表达。流式细胞术检测DADS与Chk2沉默BGC823细胞周期分布情况。Western blot检测Cdc25C与cyclin B1表达。流式细胞术显示,DADS作用BGC823细胞后,G2/M期细胞呈时间依赖性增加 (P<0.05)。Chk2沉默BGC823细胞Chk2 mRNA与蛋白表达下调 (P<0.05)。Chk2沉默组G2/M期细胞较对照组差异无显著性 (P>0.05)。DADS处理Chk2沉默组与对照组后,两者G2/M期细胞差异无显著性(P>0.05),而较未处理前有明显差异 (P<0.05)。Chk2沉默后,CDC25C和CyclinB1表达较对照组无明显差异 (P>0.05)。但是, DADS处理对照组与Chk2沉默组后,CDC25C和CyclinB1表达较处理前明显降低 (P<0.05)。表明Chk2 沉默对DADS阻滞BGC823细胞G2/M作用没有影响,DADS阻滞G2/M的检查点可能不是Chk2。

Silencing Chk2 gene impact the G2/M in gastric cancerBGC823 cells by DADS

Cell cycle checkpoint kinase 1/2(chk1/2) plays an important role in the G2/M phase. This study is to investigate the effect of silent Chk2 gene on the G2/M arrest in gastric cancer BGC823 cells induced by diallyl disulfide (DADS). Using RNAi technology to silence the Chk2 gene, q-PCR and Western blot were used to detect Chk2 mRNA and protein expression. Flow cytometry was used to detect the cell cycle distribution of DADS and silencing Chk2. The expression of Cdc25C and Cyclin B1 was detected by Western blot. flow cytometry showed that the cells of G2/M increased in a time-dependent manner after DADS treated BGC823 cells (P<0.05). After silence Chk2 gene, Chk2 mRNA and protein expression were down-regulated (P<0.05). There was no significant difference in the G2/M phase cells in the Chk2 silent group (P>0.05). There was no significant difference between the G2/M cells in the G2/M period after DADS treated Chk2 silent group and the control group (P>0.05), while there was a significant difference before the treatment (P<0.05). After silence Chk2, the expression of CDC25C and CyclinB1 was not significantly different from that in the control group (P>0.05). However, the expression of CDC25C and CyclinB1 was significantly lower than that before treatment after DADS treated control group and Chk2 silent group (P<0.05). It indicates that Chk2 silence had no effect on the G2/M effect of DADS blocking BGC823 cells, and the checkpoint for DADS block G2/M may not be Chk2.