siRNA沉默C-erbB-2基因对人喉癌Hep-2细胞PI3K/Akt信号通路的影响

The effects of C-erbB-2 siRNA on PI3K/Akt signaling pathway in human laryngeal cancer Hep-2 cells

目的:探讨siRNA沉默人喉癌Hep-2细胞C-erbB-2基因对细胞增殖的影响,及其与PI3K/Akt信号通路的关系。方法:利用siRNA技术转染Hep-2细胞C-erbB-2基因,RT-PCR3检测siRNA转染后C-erbB-2mRNA的水平变化,MTT检测细胞增殖情况改变,Western blot检测ERK1/2和AKT1蛋白磷酸化变化情况。结果:转染C-erbB-2siRNA 24h后的Hep-2细胞C-erbB-2mRNA水平出现下调,细胞增殖抑制率增加,ERK1/2和AKT1蛋白磷酸化水平也相应降低,C-erbB-2基因表达与ERK1/2和AKT1蛋白磷酸化水平之间有显著相关性。结论:siRNA沉默Hep-2细胞C-erbB-2抑制Hep-2细胞增殖,这种增殖抑制可能通过降低ERK1/2和AKT1蛋白磷酸化水平实现。

Objective：To investigate the effects of short interfering RNA（siRNA） silencing C-erbB-2 gene on the proliferation of laryngeal carcinoma Hep-2 cells and to explore the relationship between C-erbB2 gene and pI3K/Akt signal pathwayl Method：C-erbB-2 siRNA was transfected to Hep-2 cells,and mRNA expression of C- erbB-2 was detected by RT-PCR. MTT was applied to detect the proliferation of Hep-2 cells. Western blot was used to detect the phosphorylation changes of protein ERK1/2 and AKT1. Result：The mRNA level of C-erbB-2 in Hep-2 cells declined obviously at 24 h after transfection of C-erbB-2 siRNA. The proliferation of transfected Hep- 2 cells was inhibited, and protein phosphorylation levels of ERK1/2 and AKT1 decreased simultaneously. The expression of C-erbB-2 gene was obviously correlated with the protein phosphorylation levels of ERK1/2 and AKT1. Couclusion：C-erbB-2 silence by siRNA can efficiently inhibit the proliferation of Hep-2 cells, which may be a- chieved by decreasing the phosphorylation level of ERK1/2 and AKT1.