摘要
目的:探讨Chk1基因在二烯丙基二硫诱导的乳腺癌细胞MCF-7凋亡和细胞周期中的作用。方法:采用siRNA转染MCF-7细胞靶向干扰Chk1基因,采用Western blot检测Chk1蛋白表达;采用流式细胞术检测Chk1基因沉默对二烯丙基二硫诱导乳腺癌细胞凋亡及细胞周期的影响。结果:转染Chkl siRNA后,乳腺癌细胞MCF-7中Chkl蛋白表达水平降低,明显低于对照组(P<0.05)。FCM检测结果显示,转染Chkl siRNA后,G2/M期乳腺癌细胞数量有所降低(P<0.05),并且si-Chkl+DADS组G2/M期比率明显低于NC siRNA+DADS组(P<0.05)。Chk1基因沉默后,二烯丙基二硫诱导的细胞凋亡率由(14.7±1.0)%上升到(29.8±1.7)%。结论:siRNA抑制Chk1基因表达可明显消除G2/M期阻滞,并显著增强二烯丙基二硫诱导乳腺癌细胞MCF-7凋亡的敏感性。
Objective To investigate the effects of Chkl gene on diallyl disulfide- induced apopotosis and cell cycle in MCF-7 cell line. Methods The siRNA targeting against Chk1 gene was transfected into MCF-7 cells. The protein expression of Chk1 was detected by Western blotting assay. Cell cycle and apoptosis was determined by flow cytometry. Results The protein level of Chk1 was significantly reduced in Chk1 siRNA-transfected group compared with that of control group( P < 0.05). The percentage of the si-Chk1 group cells in G2/ M phase was lower than that in control group( P < 0. 05). The percentage of the si-Chk1 + DADS group cells in G2/ M phase was lower than NC siRNA + DADS group( P < 0. 05). Chk1silence increased MCF-7 apoptosis( 14. 7 ± 1. 0% vs. 29. 8 ± 1. 7%). Conclusion Chk1 silence decreased G2/ M arrest and sensitized diallyl disulfide-induced apoptosis in MCF-7 cells.
出处
《湖北医药学院学报》
CAS
2013年第5期372-376,共5页
Journal of Hubei University of Medicine
