HIF-1α基因干扰对大鼠CBRH-7919肝癌细胞HIF-1α与VEGF表达影响的研究  

Effects of HIF-1α siRNA interference on expressions of HIF-1α and VEGF in rat CBRH-7919 hepatoma cells

作　　者：倪嘉延[1] 吴裕丹[2] 黄康华[1] 许林锋[1] 

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

出　　处：《中华肿瘤防治杂志》2012年第22期1704-1708,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：广东省自然科学基金(10151008901000182)

摘　　要：目的:探讨沉默缺氧诱导因子-1α(HIF-1α)对肝癌细胞在缺氧状态下表达HIF-1α及血管内皮生长因子(VEGF)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)模拟缺氧状态,构建3组特异性较强的HIF-1αsiRNA表达载体,采用小分子RNA干扰技术沉默细胞HIF-1α的基因表达,分别利用real timeRT-PCR和蛋白质印迹法检测肝癌细胞HIF-1α和VEGF在mRNA及蛋白水平的表达变化。结果:在不同浓度CoCl2模拟缺氧条件下,随着CoCl2浓度的增高及时间的推移,肝癌细胞HIF-1α和VEGF的表达(mRNA及蛋白水平)较常氧对照组(0μmol/L CoCl2)显著增多,差异均有统计学意义,P<0.05。应用3组不同HIF-1α和siRNA表达载体转染缺氧条件下的肝癌细胞后,细胞的HIF-1α和VEGF的基因(mRNA)表达量与未转染HIF-1αsiRNA表达载体对照组相比较明显减少,同步检测两者的蛋白表达亦明显减少,与对照组差异均有统计学意义,P<0.05。结论:缺氧可以诱导肝癌细胞HIF-1α和VEGF的表达,siRNA沉默HIF-1α能显著抑制缺氧状态下诱导的VEGF表达。OBJECTIVE： To study the expression of hypoxia inducible factor-1a （HIF-1a） in rat CBRH-7919 hepatoma cells under hypoxia,and explore the effects of silencing HIF-ltx on expressions of HIF-1a and vascu1ar enothe1al growth factor （VEGF）. METHODS： Rat CBRH-7919 hepatoma cells line was used for this study and the hypoxia model was constructed by COC12. Under hypoxia,the expressions of HIF-1a and VEGF in rat CBRH-7919 hepatoma cells were detected by RT-PCR and Western-Blot. According to the HIF-1a gene sequence, HIF-1 a specific RNAi sequence was constructed and transfected into cells. After the transfection of siRNA,RT-PCR and Western blot analysis were used to detect the expressions of HIF-1a and VEGF rnRNA and protein, respectively. RESULTS： Under different density of COC12 caused hypoxia, as a result of the increased density of COC12 and the prolong of exposed time,the expressions of HIF-1a and VEGF mRNA and protein were significantly increased with statistic significance（P%0.05）. After three different groups of H IF-1a siRNA were transfected, the expressions of H IF-1a and VEGF mRNA and protein were significantly decreased compared with control group with statistic significance（P〈0.05）. CONCLUSIONS： Hypoxia can induce the expressions of HIF-1a and VEGF in Rat CBRH-7919 hepatoma cell line. HIF-1a silencing can significantly reduce the expressions of HIF-1a and VEGF.

关 键 词：肝肿瘤 缺氧诱导因子-1Α RNA干扰 血管内皮生长因子 大鼠 

分 类 号：R735.7[医药卫生—肿瘤]