Yin Yong-Shuo, Xiao Qiang, Xie Yu-Bo, Li Lei, Wang Chang-Qing, Ma Yu-Lin, Tang Zheng-Yong
Department of Gastrointestinal and Gland Surgery, The First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi, 530027, PR China .
Ai Zheng. 2008 Sep;27(9):914-8.
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.
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.
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)
E2F-1 siRNA could down-regulate E2F-1 expression in human gastric cancer MGC803 cells and suppress its invasion and proliferation to some extent.
转录因子E2F-1作为细胞周期的重要调控因子,与肿瘤发生密切相关。本研究旨在探讨E2F-1小干扰RNA(siRNA)对人胃癌MGC803细胞侵袭和增殖的影响。
将含短发夹结构的E2F-1 siRNA载体转染至MGC803细胞。未转染细胞和pSilencer4.1阴性对照转染细胞作为对照。采用逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹法检测E2F-1的表达。采用细胞基质胶侵袭实验和克隆实验检测E2F-1 siRNA转染后MGC803细胞的侵袭和增殖能力。
E2F-1 siRNA转染48小时后,E2F-1的mRNA水平较对照下调超过90.0%;E2F-1的蛋白水平较阴性对照下调79.6%,较空载体对照下调81.5%。E2F-1 siRNA组迁移细胞数明显少于阴性对照和空载体对照组(18.0±2.6 vs. 48.0±4.6和54.0±5.6,P<0.05)。E2F-1 siRNA组细胞克隆数也明显少于阴性对照和空载体对照组(46.0±2.0 vs. 122.3±1.5和128.7±2.1,P<0.05)。
E2F-1 siRNA可下调人胃癌MGC803细胞中E2F-1的表达,并在一定程度上抑制其侵袭和增殖。