摘要
目的:探索通过RNA干涉技术抑制gankyrin的表达对肝癌细胞系HepG2增殖及细胞周期的影响.方法:建立稳定表达靶向gankyrin的shRNA的HepG2细胞系.通过Westernblotting验证基因表达的抑制效果.采用细胞计数及MTT法检测细胞的增殖能力.流式细胞仪分析细胞周期.结果:抑制gankyrin基因的表达显著抑制了HepG2细胞的生长,与转染阴性对照shRNA质粒细胞相比,细胞计数示细胞数目明显减低(d6:24.4×103±5.2×103vs123.3×103±2.8×103,P<0.05),MTT分析显示细胞增殖率明显下降(144h:7.53±0.50vs16.30±0.38,P<0.05),流式细胞分析示细胞周期阻滞在G1期(G1:71.63±3.60%vs52.57±2.82%,P<0.05).结论:在HepG2细胞中抑制gankyrin的表达可有效抑制细胞生长并导致细胞周期的阻滞.利用RNA干涉技术抑制gankyrin的表达可能会成为一种有效的肝癌基因治疗手段.
AIM: To explore the effect of suppressing gankyrin expression using RNA interference (RNAi) technique on the proliferation and cell cycle of hepatocellular carcinoma cell line HepG2. METHODS: HepG2 cell line stably expressing shRNA targeted at gankyrin gene was established. The gankyrin expression was assessed by Western blotting analysis. Cell growth was evaluated by cell counting and MTT assay, and cell cycle was analyzed by flow cytometry. RESULTS: The growth of HepG2 cells was inhibited significantly by suppressing the expression of gankyrin. Compared with the cells transfected with control plasmid, the number of the gankyrin-suppressed cells was much lower (d 6:24.4×103±5.2×103 vs 123.3×103±2.8×103, P<0.05). MTT assay showed cell proliferation was decreased (144 h: 7.53±0.50 vs 16.30±0.38, P<0.05), flow cytometric analysis demonstrated that the cell cycle was stuck at G1 phase (G1: 71.63±3.60% vs 52.57±2.82%, P<0.05). CONCLUSION: Suppression of the expression of gankyrin in HepG2 cells leads to cell proliferation inhibition and cell cycle arrest. RNAi-mediated inhibition of gankyrin may be a useful therapeutic approach for human hepatocellu-lar carcinoma.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第8期988-992,共5页
World Chinese Journal of Digestology
