ZNF278 siRNA抑制胃癌细胞株AGS增殖的实验研究  

Experimental Study on Suppression of Cell Proliferation in Gastric Cancer Cell Line AGS by ZNF278 siRNA

作　　者：田筱青[1] 孙丹凤[1] 房静远[1] 

机构地区：[1]上海交通大学医学院附属仁济医院消化科 上海市消化疾病研究所,200001

出　　处：《胃肠病学》2013年第12期724-727,共4页Chinese Journal of Gastroenterology

摘　　要：ZNF278属C2H2型锌指蛋白,为参与生长发育的重要转录因子,其异常表达可能参与肿瘤发生。目的:研究ZNF278 siRNA对胃癌细胞株AGS增殖和周期的影响。方法:以蛋白质印迹法检测正常胃上皮细胞株GES-1和胃癌细胞株AGS中ZNF278表达。构建ZNF278 siRNA片段,并转染胃癌AGS细胞,转染阴性对照siRNA和RPMI1640分别作为阴性对照组和空白对照组。以定量RT-PCR和蛋白质印迹法分别检测ZNF278 mRNA和蛋白表达,利用MTT和细胞计数法检测细胞增殖情况,流式细胞术检测细胞周期变化。结果:胃癌AGS细胞中ZNF278表达明显高于正常胃上皮细胞GES-1。ZNF278 siRNA转染胃癌AGS细胞后,ZNF278 mRNA和蛋白表达均明显下调,而阴性对照组与空白对照组无明显差异。MTT法示ZNF278 siRNA组AGS细胞增殖率显著低于阴性对照组(0.64±0.03对0.81±0.03,P<0.01),细胞计数法示细胞数量亦显著降低[(4.11±0.35)×105对(5.78±0.50)×105,P<0.01],流式细胞术显示ZNF278 siRNA组G0/G1期细胞明显增加而S期细胞明显减少(P<0.05)。结论:转染ZNF278 siRNA可抑制胃癌AGS细胞增殖,细胞周期阻滞于G0/G1期。Background： ZNF278 is a novel Krtippel Cys2-His2-type zinc finger protein. It is an important transcription factor involved in growth and development. Abnormal expression of ZNF278 might be involved in tumorigenesis. Aims： To study the influence of ZNF278 siRNA on cell proliferation and cell cycle in gastric cancer cell line AGS. Methods： ZNF278 expression in human gastric epithelial cell line GES-1 and gastric cancer cell line AGS was detected by Western blotting. ZNF278 siRNA was constructed and transfected into AGS cells. Transfection with negative siRNA and RPMI1640 were served as negative controls and blank controls, respectively. Expressions of ZNF278 mRNA and protein in AGS cells were determined by quantitative RT-PCR and Western blotting, respectively. Cell proliferation was assessed by MTT assay and cell counting assay. Cell cycle was analyzed by flow eytometry. Results： The expression of ZNF278 was higher in AGS cells than in GES-1 cells. Expressions of ZNF278 mRNA and protein were significantly decreased in ZNF278 siRNA group than in negative controls. No significant difference was found between negative controls and blank controls. MTT assay showed that cell proliferation in ZNF278 siRNA-transfected AGS cells was significantly lower than that in siRNA negative controls （0.64±0.03 vs. 0. 81 -0.03, P 〈 0.01 ）. Cell counting assay showed that the number of cell was significantly lower in AGS cells transfected with ZNF278 siRNA than in siRNA-negative controls [ （4.11±0.35 ） × 10 5 vs. （5.78±0. 50） × 105 , P 〈 0.011. Flow cytometry showed that ZNF278 siRNA transfection significantly increased the percentage of GO/Gl-phase cells and decreased the percentage of S-phase cells （ P 〈 0.05）. Conclusions ： ZNF278 siRNA transfection results in a significant inhibition of gastric cancer cell proliferation and blocking the cell cycle at GO/Gl-phase.

关 键 词：ZNF278 RNA 小分子干扰 细胞增殖 细胞周期 胃肿瘤 

分 类 号：R735.2[医药卫生—肿瘤]