RNA 干扰人永生化食管上皮恶变细胞中核干细胞因子对PI3K／AKT表达的影响

摘要

Aim: To explore the effect of inhibiting nucleostemin (NS) on PI3K/AKT signal pathway in SHEEC. Methods:RNA interfering agents ( NS-RNAi) was transfected into SHEEC to inhibit the expression of NS gene , with the SHEEC group, negative control(NC) group, and MOCK group as control, and the expressions of PI3K/AKT pathway relat-ed factors were detected by using RT-PCR and Western blot methods .Results: SHEEC was transfected by NS-RNAi and the transfection rate was high.The expression levels of PI3K and AKT mRNA[(0.460 ±0.207) and (0.492 ±0.233)] were obviously down-regulated by inhibiting NS gene in comparison with SHEEC , NC and MOCK groups(P<0.05).The expression levels of PI3K and AKT proteins were (0.353 ±0.361) and (0.312 ±0.068), and showed significant differ-ence in comparison with SHEEC, NC and MOCK groups(P<0.05).Conclusion: The expressions of PI3K/AKT signal pathway related factors decrease with NS down-regulation in SHEEC , and the PI3K/AKT signal pathway may be a kind of NS P53-independent pathway .%目的：研究抑制人永生化食管上皮恶变细胞（SHEEC）中核干细胞因子（NS）对PI3K／AKT信号转导通路中相关分子表达的影响。方法：将干扰试剂（ NS-RNAi）转染至SHEEC中以抑制NS的表达，以SHEEC组、阴性对照组（NC组）和MOCK组为对照，采用RT-PCR和Western blot方法检测干扰后PI3K／AKT通路中相关因子mR-NA和蛋白表达水平的变化。结果：NS-RNAi成功转染至SHEEC 中。抑制SHEEC 中NS的表达后PI3K和AKT mRNA的相对表达量为（0．460±0．207）和（0．492±0．233），与SHEEC组、NC组和MOCK组比较，差异有统计学意义（P均＜0．05）；干扰NS表达后PI3K和AKT蛋白的相对表达量为（0．353±0．361）和（0．312±0．068），与SHEEC组、NC组和MOCK组比较，差异有统计学意义（P均＜0．05）。结论：抑制SHEEC中NS表达后PI3K／AKT信号通路中相关因子的表达均明显下降，PI3K／AKT信号通路可能是NS发挥作用的一种非依赖P53作用的信号途径。