转录因子E2F-1 siRNA对胃癌MGC803细胞体外侵袭及增殖能力的影响  

Inhibitory Effects of Transcription Factor E2F-1 siRNA on Invasion and Proliferation of Gastric Cancer Cell Line MGC803

作　　者：尹永硕[1] 肖强[1] 谢玉波[2] 李雷[1] 王长青[1] 马玉林[1] 唐振勇[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,广西南宁530027 [2]广西医科大学第一附属医院麻醉科,广西南宁530027

出　　处：《癌症》2008年第9期914-918,共5页Chinese Journal of Cancer

基　　金：广西自然科学基金项目(No.0640085);广西医疗卫生重点科研课题(No.200625)~~

摘　　要：背景与目的:转录因子E2F-1作为细胞周期进程中重要的调控因子,与肿瘤的发生有着密切的关系。本研究通过转染人E2F-1基因的小干扰RNA(small interfering RNA,siRNA)表达质粒,探讨其对胃癌MGC803细胞中E2F-1表达及侵袭和增殖能力的影响。方法:用脂质体LipofectamineTM2000将具有短发夹结构的人E2F-1siRNA表达质粒转染至MGC803细胞,以逆转录-聚合酶链反应(RT-PCR)及蛋白印迹(Western blot)技术分别检测E2F-1mRNA及蛋白的表达;应用体外侵袭实验及克隆实验分别检测E2F-1 siRNA表达质粒对MGC803细胞侵袭力和增殖能力的影响。结果:成功转染E2F-1 siRNA表达质粒48h后,MGC803细胞中E2F-1基因mRNA表达的抑制率超过90%;与未转染组及阴性对照组细胞比较,E2F-1蛋白的表达分别降低81.5%和79.6%。转染pSilencer4.1-E2F-1质粒组的穿膜细胞数(18.0±2.6)与阴性对照组(48.0±4.6)和空白对照组(54.0±5.6)比较差异有统计学意义(P<0.05),其细胞集落形成数(46.0±2.0)与阴性对照组(122.3±1.5)和空白对照组(128.7±2.1)比较差异也有统计学意义(P<0.05)。结论:E2F-1siRNA表达质粒可显著下调E2F-1基因在胃癌MGC803细胞中的表达,在一定程度上抑制胃癌细胞的侵袭和增殖。BACKGROUND ＆ OBJECTIVE. As an important regulatory factor of cell cycle, transcription factor E2F-1 is closely related to tumorigenesis. This study was to investigate the effects of E2F-1 small interfering RNA （siRNA） on invasion and proliferation of human gastric cancer MGC803 cells. METHODS： E2F-1 siRNA vector containing short hairpin structure was transfected into MGC803 cells. Untransfected and pSilencer4.1-negative-transfected cells were used as controls. The expression of E2F-1 was detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot. Cell matrigel invasion assay and cloning assay were used to detect the invasion and proliferation of MGC803 cells after E2F-1 siRNA transfection. RESULTS： At 48 h after E2F-1 siRNA transfection, the mRNA level of E2F-1 was down-regulated by over 90.0% of controls; the protein level of E2F-1 was down-regulated by 79.6% of negative control and by 81.5% of empty control. The number of migrated cells was significantly smaller in E2F-1 siRNA group than in negative and empty control groups （18.0±2.6 vs. 48.0±4.6 and 54.0±5.6, P〈0.05）. The number of cell clones was also significantly smaller in E2F-1 siRNA group than in negative and empty control groups （46.0±2.0 vs. 122.3±1.5 and 128.7±2.1, P〈0.05） CONCLUSION. E2F-1 siRNA could down-regulate E2F-1 expression in human gastric cancer MGC803 cells and suppress its invasion and proliferation to some extent.

关 键 词：RNA干扰 E2F-1 胃肿瘤 MGC803细胞 侵袭 增殖 

分 类 号：R735.2[医药卫生—肿瘤]