摘要
目的筛选NS基因特异性siRNA阳性细胞克隆,研究NS基因特异性RNA干扰对Eca-109细胞株体外增殖能力的影响。方法用脂质体法将NS特异性siRNA表达载体转染Eca-109细胞,Zeocin抗生素压力筛选后用PCR方法鉴定阳性克隆;MTT法检测各组细胞的生长增殖情况,RT-PCR方法检测NS基因表达量的变化情况。结果与对照组比较,silencer组肿瘤细胞趋于分化,细胞增殖抑制率超过80%(P<0.01),差异具有显著性;NS基因表达量下降。结论NS基因特异性RNA干扰抑制人食管癌Eca-109细胞株体外增殖,使NS基因表达量下降。
Objective To screen NS gene specific positive cell clones,and investigate the effect of human esophagus cancer Eca-109 cells proliferation in vitro by NS gene specific RNA interference. Methods The NS-siRNA expression vector was transfected into human esophagus cancer Eca-109 cells using LIPOFECTAMINEru2000 Reagent, and positive clones were examined by PCR after stable transfectans screening with Zeocin; detected the cell proliferation by MTT assay and the levels of NS gene expression by RT-PCR. Results Compared with the two control groups, the silencer group calls were nearer differetiation, the proliferation inhibitory rate was higher than 80% （P〈0. 01）; the level of NS gene expression reduced. Conclusion The NS gene specific RNAi can significantly inhibit human esophagus cancer Eca- 109 cells proliferation in vitro and reduce the level of NS gene expression.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2007年第2期103-105,共3页
Cancer Research on Prevention and Treatment
基金
牡丹江医学院科研项目
