摘要
目的观察周期蛋白依赖性激酶1(CDK1)在胃癌细胞及永生化胃粘膜上皮细胞系中的表达,探讨其沉默对胃癌细胞恶性表型的影响。方法利用实时荧光定量PCR检测永生化胃粘膜上皮细胞系GES及胃癌细胞系SGC7901中CDK1的mRNA表达水平,应用RNA干扰技术沉默胃癌细胞系SGC7901的CDK1,运用流式细胞术检测沉默CDK1前后胃癌细胞的凋亡情况,绘制细胞的生长曲线,并通过克隆形成实验观察肿瘤细胞恶性表型的变化。结果 CDK1在胃癌细胞系SGC7901中表达明显高于永生化胃粘膜上皮细胞系GES。使用siRNA体外转染SGC7901细胞后胃癌细胞的凋亡率升高。MTT结果显示与空白对照组和阴性对照相比,沉默CDK1后胃癌细胞增殖速度明显受到抑制(P<0.05)。克隆形成实验发现沉默CDK1后可明显减少胃癌细胞株的平板克隆数目。结论 CDK1在胃癌细胞中高表达,可能参与胃癌的发生发展,下调CDK1的表达可促进胃癌细胞凋亡、抑制胃癌细胞的恶性增殖。
Objective The purpose of this study was to investigate the expression of cyclin dependent kinase 1(CDK1)in gastric cancer cell line SGC7901 and gastric epithelial cell line GES and study the effects of CDKl knockdown on the malignant be-havior of gastric cancer. Methods The expression of CDKl in gastric epithelial cell line GES and gastric cancer cell line SGC7901 were quantified using real-time quantitative polymerase chain reaction( Q-PCR) . The siRNA targeting the CDK1 gene was synthe-sized and transfected into SGC7901 cells by lipofectamine 2000. Cell apoptosis was detected by flow cytometry after the transfected for 48 or 60 h. MTT assay and colony formation assay were performed to examine the effect of CDKl on the cell growth and cell proliferation in vitro. Results CDK1 expression was significantly increased in gastric cancer cell line SGC7901 compared with gastric epithelial cell line GES. The ratio of apoptotic cell of transfected cells at 48h after transfection was increased significantly compared with control. Downregulation of CDK1 inhibited the growth and proliferation of gastric cancer cell compared with control, which was comfirmed by MTT and plate colony formation assays. Conclusion CDK1 expression was significantly increased in gas-tric cancer. Overexpression of CDK1 may promote oncogenesis and progression of human gastric cancer. Downregulation of CDK1 expression can prompt the apoptosis and inhibit the proliferation activities of human gastric cancer.
出处
《四川医学》
CAS
2015年第6期827-830,共4页
Sichuan Medical Journal
关键词
胃癌
CDK1
凋亡
增殖
gastric cancer
CDKI
apoptosis
proliferation