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*
RNA interfering effect of nucleostemin on expression of PI3K/AKT in Shantou human embryonic esophageal carcinoma epithelial cells

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mRNA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。
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 related factors were detected by using RT-PCR and Western blot methods.Results: SHEEC was transfected by NS-RNAiand 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 difference 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