摘要
目的探讨干扰Timeless表达对SGC-7901胃癌细胞凋亡、周期及β-连环蛋白(β-catenin)表达的影响。方法合成Timeless的siRNA,通过Lipofectamine?3000转染SGC-7901细胞,检测并选择干扰效率高的siRNA用于Timeless干扰组细胞造模。设置分组:空白组,siRNA-对照组和siRNA-Timeless组。Realtime PCR及western blot法检测Timeless、β-catenin mRNA及蛋白表达,MTT法检测细胞活力,流式细胞术检测细胞凋亡及细胞周期。结果Realtime PCR及western blot法均证实Timesless-siRNA-2转染效率最高。与空白组及siRNA-对照组比较,siRNA-Timeless组Timeless、β-catenin mRNA及蛋白表达量下调(P<0.01),细胞活力降低(P<0.01),细胞凋亡率升高(P<0.01),细胞周期阻滞于S期(P<0.01)。结论干扰Timeless表达能显著诱导SGC-7901胃癌细胞凋亡,抑制细胞增殖,与下调β-catenin表达有关。
Objective To explore effects of Timeless interference on apoptosis,cycle and expression of β-catenin in SGC-7901 gastric cancer cells.Methods The siRNA of Timeless was synthesized and transfected into SGC-7901 cells by Lipofectamine?3000.Then the most efficient Timeless siRNA was chosen to make the Timeless interference group.Three experiment groups were set:blank control,siRNA control group and siRNA-Timeless group.The expression of Timeless and β-catenin mRNA and protein was measured by Realtime PCR and western blot.Cell viability was measured by MTT.Cell apoptotic rate and cell cycle was detected by flow cytometry.Results The transfection efficiency of Timesless-siRNA-2 was the highest via Realtime PCR and western blot.Compared with blank control group and siRNA control group,cell viability was decreased(P<0.01),cell apoptotic rate was increased(P<0.01),cell cycle was made arrested at S phage(P<0.01),the expression of β-catenin was down-regulated in siRNA-Timeless group(P<0.01).Conclusion Interference with Timeless expression could significantly induce apoptosis of SGC-7901 gastric cancer cells,and inhibit cell proliferation,which related to down-regulation expression of β-catenin expression.
作者
李翔
Li Xiang(Medical Department of Nanchang University,Nanchang,Jiangxi,330000,China;Department of Oncology,Jiangxi Cancer Hospital,Nanchang,Jiangxi,330000,China)
出处
《当代医学》
2020年第9期83-87,共5页
Contemporary Medicine
