Hec1基因表达沉默对胃癌细胞凋亡及分裂周期的影响

目的研究癌症高表达(Hec1)基因表达沉默对胃癌细胞AGS分裂周期及细胞凋亡的影响。方法应用siRNA使Hec1基因表达下调,通过实时定量PCR检测Hec1mRNA表达抑制率;流式细胞仪检测细胞周期变化,经AnnexinV/PI双染色检测细胞凋亡率。结果胃癌细胞AGS转染Hec1siRNA48h后,Hec1基因表达抑制率达到90%;流式细胞仪检测发现,细胞G2/M期比例显著高于各对照组(P<0.05),细胞分裂被阻滞,出现明显凋亡;形态学检测发现,Hec1siRNA转染48h后处于分裂中期细胞高于各对照组,转染72h后细胞呈明显凋亡形态学改变。AnnexinV/PI双染色法显示Hec1基因沉默后细胞凋亡率高于对照(P<0.05)。结论抑制Hec1基因表达,可以明显阻滞细胞分裂并停留在有丝分裂中期,同时诱导细胞凋亡。

Effect of Hec1 Gene Silencing on Cell Cycle Arresting and Apoptosis

Objective To study the effect of high expression in cancer(Hec1) gene silencing on cell cycle arresting and apoptosis. Methods Hec1 gene expression was silenced by siRNA transfection, and the gene expression level was detected by real-time quantitative PCR. Apoptotic cell and DNA contents were measured by flow cytometry 48 h after Hec1 siRNA transfection. Morphological changes were observed under the light microscope. Results Forty-eight hours after Hec1 siRNA transfection, the suppression rate of Hec1 gene expression reached 90%.The cells with Hec1 gene silenced had higher ratio of G_2/M compared with control group (P<0.05). By light microscopy, it was found that there were more mitotic phase and apoptotic cells in Hec1 gene silence group than those in control group. High apoptosis rate in Hec1 gene silenced cells was also found by Annexin V/PI double staining. Conclusion Hec1 gene silence may lead to cell cycle arrest in mitotic phase and cell apoptosis.