摘要
目的:观察siRNA沉默Cyclin E基因表达对肝癌HepG2、SMMC-7721和BEL-7402细胞增殖和侵袭能力的影响.方法:构建2个靶向Cyclin E基因siRNA载体,转染人肝癌HepG2、SMMC-7721和BEL-7402细胞.RT-PCR、Western blot检测转染后HepG2、SMMC-7721和BEL-7402细胞Cyclin E基因mRNA和蛋白表达水平.CCK-8试验、软琼脂克隆形成实验检测HepG2、SMMC-7721和BEL-7402细胞增殖、克隆形成能力.流式细胞术、transwell试验分别检测HepG2、SMMC-7721和BEL-7402细胞周期和侵袭能力.结果:构建的2个Cyclin E基因siRNA载体插入序列与所设计序列均一致;转染HepG2、SMMC-7721和BEL-7402细胞后,干扰1组、干扰2组与空白对照组和阴性对照组比较,C y c l i n E m R N A和蛋白表达量均显著降低(P<0.05),细胞生长速度延缓,软琼脂细胞集落形成数、穿透细胞数均显著降低(P<0.05),S和G2/M期细胞比例减少,G0/G1期细胞比例增加.结论:沉默肝癌细胞Cyclin E表达水平,可有效抑制细胞生长、增殖和侵袭能力.
AIM： To observe the effect of RNAi-mediated silencing of the Cyclin E gene on the proliferation and invasion of HepG2,SMMC-7721 and BEL-7402 cells.METHODS： Two vectors carrying siRNA targeting the Cyclin E gene were constructed and transfected into HepG2,SMMC-7721 and BEL-7402 cells.The mRNA and protein expression of Cyclin E were measured by RT-PCRand Western blot,respectively.CCK-8 assay and colony formation assay were employed to assess the proliferation and colony-forming ability of transfected HepG2,SMMC-7721 and BEL-7402.Flow cytometry（FCM） and transwell migration assay were used to evaluate cell cycle progression and migration of transfected cells.RESULTS： Compared to the blank control group and negative control group,the expression of Cyclin E mRNA and protein was significantly decreased,the proliferation,colonyforming ability,and migration were suppressed significantly,and cell cycle was arrested in G 0 / G 1 phase in two experimental groups.CONCLUSION： Down-regulation of Cyclin E expression significantly inhibits the proliferation and migration of HepG2,SMMC-7721 and BEL-7402 cells.
出处
《世界华人消化杂志》
CAS
北大核心
2012年第22期2086-2091,共6页
World Chinese Journal of Digestology
