Nucleostemin基因沉默对食管癌细胞增殖和凋亡的影响

Effect of Nucleostemin gene silencing by siRNA on proliferation and apoptosis of esophageal cancer Eca-109 cells

目的探讨RNA干扰沉默Nucleostemin(NS)基因后对食管癌Eca-109细胞株增殖和凋亡的影响。方法用NS基因特异性siRNA表达载体转染食管癌Eca-109细胞株,通过绘制细胞生长曲线和克隆形成实验检测细胞增殖能力;CCK-8法测试细胞增殖抑制率;RT-PCR法检测NS基因表达量的变化,TUNEL法和DNA Ladder检测细胞凋亡情况。结果与对照组相比,S组细胞的增殖速率降低,24、48及72h细胞增殖抑制率分别为60.32%、72.29%和86.00%;NS基因表达量下降;细胞凋亡指数增加(18.28%),可见梯状DNA降解带。结论 NS基因沉默导致食管癌Eca-109细胞株增殖抑制,凋亡增加。

Objective To investigate the effect of Nucleostemin （NS） gene silencing by siRNA on proliferation and apoptosis of esophageal cancer Eca-109 cells.Methods The NS-siRNA expression vector was transfected into esophageal cancer cells with LipofectamineTM2000 reagent.Cell proliferation was assessed by observing the growth curve and clone formation of the cells.The rate of cell proliferation inhibition was detected by CCK-8 assay.Level of NS gene expression in all groups was measured by RT-PCR.Cell apoptosis was examined by TUNEL kit and DNA ladder.Results After cultured 24,48 and 72 h,cell proliferation reduced,the rate of cell proliferation inhibition in S group was 60.32%,72.29% and 86.00%.The level of NS gene expression reduced in S group,the cell apoptosis index increased and showed a typical DNA Ladder on the gel.Conclusions NS gene silencing induces esophageal cancer Eca-109 cells proliferation inhibition and increases cell apoptosis.