RNA干扰沉默乏氧诱导因子-1α对紫杉醇诱导食管鳞癌EC9706细胞凋亡的影响  

Effects of silencing HIF-1α gene by RNA interference on Taxol-induced apoptosis of human esophageal squamous cell line EC9706

作　　者：吴欣爱[1] 孙燕[2] 樊青霞[1] 王留兴[1] 王瑞林[1] 张岚[3] 

机构地区：[1]郑州大学第一附属医院肿瘤科,郑州450052 [2]中国医学科学院肿瘤医院内科,北京100078 [3]郑州大学第一附属医院病理科,郑州450052

出　　处：《肿瘤》2007年第9期710-714,共5页Tumor

摘　　要：目的:探讨食管癌乏氧耐药机制,观察乏氧诱导因子-1α(HIF-1α)对紫杉醇诱导的人食管鳞癌EC9706细胞凋亡的影响。方法:用CoCl2化学乏氧法建立乏氧模型;应用Western blot测定RNA干扰前后,乏氧条件下HIF-1α蛋白表达的变化;应用原位末端标记TUNEL法及AnnexinⅤ/PI双标记法检测RNA干扰前后紫杉醇对乏氧培养细胞EC9706细胞凋亡的影响。结果:EC9706细胞经75μmol/L的CoCl2作用4 h后,即可诱导HIF-1α蛋白表达。RNA干扰技术能明显抑制乏氧诱导的HIF-1α蛋白表达。相同的乏氧条件下,转染HIF-1α-siRNA组细胞的凋亡率与未转染组或转染control-siRNA组相比较差异有统计学意义(P<0.05)。结论:乏氧条件下HIF-1α蛋白对紫杉醇诱导的EC9706细胞凋亡具有抑制作用。以HIF-1α为新的靶点可望提高食管癌的治疗水平。Objective： To study the mechanism of hypoxia-induced drug-resistance in esophageal squamous cell carcinoma and observe the effect of hypoxia inducible factor 1 alpha （ HIF-1α） on the Taxol-induced apoptosis of EC9706 cells. Methods. Small interference RNA （siRNA） targeting HIF-1α was constructed and transfected into human EC9706 cells. Then EC9706 cells were cultured under chemical hypoxia conditions induced by CoCl2, a chemical hypoxia inducer. The expression of HIF-1α in EC9706 cells were detected by Western blot before and after RNA interference. The effect of Taxol on apoptosis of EC9706 cells were determined by TUNEL assay and Annexin V/PI double staining. Results：The expression of HIF-1α protein was induced by CoC12 75 μmol/L after 4 h, and was significantly inhibited by siRNA-HIF-1α. The apoptosis rate of EC9706 cells induced by hypoxia were significantly higher in siRNA-HIF-1 α transfection group than those without transfection or transfected with control siRNA （P 〈 0.05 ）. Conclusion：Over-expression of HIF-1 α inhibited the apoptosis of EC9706 cells induced by Taxol under hypoxia, siRNA targeting HIF-1 α increases the therapeutic efficacy of esophageal squamous cell carcinoma.

关 键 词：食管肿瘤 RNA干扰 细胞凋亡 细胞乏氧 乏氧诱导因子-1Α 

分 类 号：R735.1[医药卫生—肿瘤]