摘要
目的:探讨沉默缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)基因表达对肝癌细胞PI3K/AKT信号转导通路的影响。方法:选用大鼠CBRH-7919肝癌细胞株作为研究对象,利用氯化钻(CoCl2)建立厌氧模型。构建HIF-1α小分子干扰RNA(small interfering RNA,siRNA)特异性载体复合物,小分子RNA干扰技术沉默肝癌细胞HIF-1α的基因表达,分别利用RT-PCR和免疫印迹法检测肝癌细胞HIF-1α、血管内皮生长因子(vascular endothelial growth factor,VEGF)、磷酸化AKT(p-AKT)、AKT和PI3K在mRNA和(或)蛋白水平的表达变化。结果:肝癌细胞在缺氧条件下表达HIF-1α较常氧对照组显著增多,差异具有统计学意义,PmRNA=0.002,P蛋白=0.003。特异性HIF-1αsiRNA表达载体转染肝癌细胞后,肝癌细胞HIF-1α(PmRNA和P蛋白均<0.001)、VEGF(PmRNA=0.001,P蛋白<0.001)和p-AKT(P蛋白<0.001)的表达明显减少;AKT和PI3K的表达显著增多,与对照组的差异均具有统计学意义,P蛋白值分别为0.032和0.020。结论:特异性siRNA干扰沉默HIF-1α基因表达可抑制肝癌细胞PI3K/AKT信号转导通路的激活。
OBJECTIVE： To study the effects of HIF-1αcsilencing on the PI3K/AKT transduction signaling pathway in hepatoma cells. METHODS：Rat CBRH-7919 hepatoma cells was used for this study and the hypoxia model was made u- sing COCl2. HIF-1α specific RNAi sequence compound was constructed and transfected into cells. After the transfection of special siRNA, real time RT-PCR and western blot analysis were used to detect the expressions of HIF 1α, VEGF, p-AKT, AKT and PI3K on mRNA and/or protein level. RESULTS：Under the COCl2 caused hypoxia,the expression levels of mRNA and protein of HIF-1α were significantly increased with statistic significance（PmRNA = 0. 002, Pprotein = 0. 003）. After the specific sequence of HIF-1α siRNA transfected,compared with control group,in experimental group the expres- sions of HIF-1α （PmRNA〈0.001,Pprotein〈0. 001）,VEGF （PmRNA =0. 001,Pprotoin〈0. 001） and p-AKT （Pproteln〈0. 001） were significantly decreased; while the expressions of AKT （Pprotoin = 0. 032） and PI3K （Pprotein = 0. 020） were obviously in- creased, there were significantly statistic differences between these two groups. CONCLUSION： HIF-1α silencing can inhib- it the activation of PI3K/AKT transduction signaling pathway in hepatoma cells.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2014年第5期356-359,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
广东省自然科学基金(10151008901000182)
关键词
肝肿瘤
缺氧诱导因子-1Α
RNA干扰
血管内皮生长因子
蛋白激酶B
liver neoplasms
hypoxia inducible factor-1α
RNA interference
vascular endothelial growth factor
protein kinase B
