干细胞生长因子反义寡核苷酸对肝癌细胞中原癌基因蛋白质c-kit及碱性成纤维细胞生长因子表达的影响

Effect of antisense oligonucleotide targeting SCF on c-kit and bFGF of hepatoma cells

目的探讨干细胞生长因子(SCF)反义寡核苷酸对肝癌HepG2细胞中原癌基因蛋白质c-kit、碱性成纤维细胞生长因子(bFGF)表达的影响及意义。方法以脂质体作为转染载体,转染SCF反义寡核苷酸于HepG2细胞。实验分三组:(1)空白对照组;(2)转染SCF反义寡核苷酸组;(3)转染SCF错义寡核苷酸组。Western blot方法检测转染前后肝癌细胞中SCF、bFGF蛋白的表达,RT-PCR检测转染前后肝癌细胞中c-kit与bFGF mRNA的表达,用流式细胞仪检测转染SCF反义寡核苷酸后HepG2细胞凋亡率。结果与转染错义寡核苷酸组相比,SCF反义寡核苷酸对HepG2细胞中c-kit、bFGF的表达有明显抑制作用,转染SCF反义寡核苷酸可明显增加HepG2细胞凋亡率(P<0.01)。结论 SCF在肝癌细胞凋亡中有重要调节作用,SCF/c-kit有可能作为PI3K/Akt信号通路的上游对肝癌细胞bFGF的表达起重要调控作用。

Objective To investigate the expression and significance of c-kit,bFGF in hepatoma cells of transfection of antisense oligonucleotide targeting SCF.Methods Optimize condition of transfection.Inhibition of SCF and bFGF protein expression was investigated by Western blot,c-kit and bFGF mRNA expression was investigated by RT-PCR in best condition of transfection of antisense oligonucleotide targeting SCF.The apoptosis rate of HepG2 cells was detected by flow cytometry in best condition of transfection of antisense oligonucleotide targeting SCF.Results Treatment with antisense oligonucleotide of SCF not only reduced the expression of SCF,c-kit and bFGF by Western blot and RT-PCR,but also increased the apoptosis of HepG2 cells(P<0.01).Conclusions SCF may take part in apoptosis regulation of hepatoma cells,SCF/c-kit may be used as PI3K/Akt signaling pathway upstream of the expression of bFGF.