沉默HIF-1α基因表达对大鼠肝癌细胞增殖的影响  

Effects of HIF-1α silencing on proliferation of rat hepatoma cells

作　　者：吴裕丹[1] 倪嘉延[2] 陈耀庭[2] 孙宏亮[2] 许林锋[2] 

机构地区：[1]中山大学孙逸仙纪念医院血液科,广东广州510120 [2]中山大学孙逸仙纪念医院介入放射科,广东广州510120

出　　处：《中国病理生理杂志》2013年第7期1207-1212,共6页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.10151008901000182)

摘　　要：目的:探讨沉默缺氧诱导因子1α(HIF-1α)基因表达对缺氧状态下肝癌细胞增殖的影响。方法:选用大鼠CBRH-7919肝癌细胞株作为研究对象,利用氯化钴(CoCl2)建立缺氧模型。制备3组特异性较强的HIF-1αsiRNA-脂质体复合物,转染于肝癌细胞。利用real-time RT-PCR、Western blotting等方法分别检测肝癌细胞HIF-1α、血管内皮生长因子(VEGF)、p21和cyclin D1在mRNA和(或)蛋白水平的表达。MTT及BrdU掺入实验检测细胞增殖的变化。结果:缺氧条件下,肝癌细胞的HIF-1α和VEGF mRNA及蛋白表达显著增多(P<0.05)。HIF-1α基因表达沉默后,HIF-1α、VEGF及cyclin D1 mRNA和(或)蛋白表达明显减少(P<0.05),p21蛋白表达明显增加(P<0.05)。HIF-1αsiRNA转染组的BrdU阳性细胞比例明显少于对照组(P<0.05)。结论:沉默HIF-1α基因表达对缺氧状态下肝癌细胞的增殖有显著抑制作用。AIM： To study the effect of hypoxia-inducible factor 1α（HIF-1α） silencing on the proliferation of hepatoma cells under hypoxia.METHODS： Rat hepatoma cell line CBRH-7919 was used in this study.Hypoxia model was established by treating the cells with cobalt chloride（CoCl2）.The expression of HIF-1α was silenced by small interference RNA.Real-time RT-PCR and Western blotting were used to detect the mRNA and / or protein expression of HIF-1α,vascular endothelial growth factor（VEGF）,p21 and cyclin D1 in CBRH-7919 cells under hypoxia.The proliferation of CBRH-7919 cells was measured by the technique of 5-bromo-2’-deoxyuridine（BrdU） incorporation.RESULTS： The expression of HIF-1α and VEGF at mRNA and protein levels was significantly increased under hypoxia（P 0.05）.Silencing of HIF-1α significantly inhibited the expression of HIF-1α,VEGF and cyclin D1 at mRNA and / or protein levels,while increased the protein expression of p21（P 0.05）.The BrdU-positive cells in HIF-1α siRNA transfection group were significantly less than those in control group.CONCLUSION： HIF-1α silencing significantly inhibits the proliferation of hepatoma cells under hypoxia.

关 键 词：缺氧诱导因子1Α RNA干扰 P21蛋白 细胞周期蛋白D1 CBRH-7919细胞 

分 类 号：R735.2[医药卫生—肿瘤]