RNA干扰缺氧诱导因子2对肾癌786-0细胞的影响

目的：研究RNA干扰缺氧诱导因子2（HIF-2）对肾癌786-0细胞的影响。方法:合成HIF-2 RNAi片段，将RNAi片段定向克隆于RNAi表达载体，测序鉴定；转染人肾癌786-0细胞, RT-PCR、Western 印迹法检测HIF-2 RNAi表达载体对HIF-2基因表达的影响，MTT法检测转染HIF-2 RNAi表达载体对细胞生长作用，裸鼠种植肿瘤观察RNA干扰组对肿瘤生长的影响。结果:RNA干扰组HIF-2mRNA的表达受到抑制，明显低于空载体组和对照组HIF-2mRNA的表达(P<0.01),空载体组HIF-2mRNA的表达没有受到影响；RNA干扰组的HIF-2蛋白表达明显降低，明显低于空载体组和对照组(P<0.01)，空载体组和对照组HIF-2蛋白的表达无显著差异；RNA干扰组细胞生长减慢,RNA干扰组肿瘤体积和瘤重均小于空白组(P<0.01)。结论:RNA干扰HIF-2可以抑制肾癌786-0细胞HIF-2的表达和抑制细胞生长，减慢裸鼠肿瘤的生长；为肾癌的生物治疗提供一定的实验基础。

Effect of RNA interference on HIF-2 in the renal cancer cell line 786-0

AIM: To study the effect of RNA interference on hypoxia-inducible factor-2 (HIF-2) in the renal cell cancer in vitro and in vivo. METHODS: HIF-2 RNAi was synthesized and inserted into RNA interference eukaryotic expression vector which was confirmed by sequencing. The vector was transfected into the renal cancer cell 786-0 and positive clone was selected by using G418. The HIF-2 expression was detected by RT-PCR and Western blotting method. The growth of cells was measured by MTT method. Nude mouse xenograft assays were also done. RESULTS: Compared with empty vector group and control group, the amounts of HIF-2 mRNA and protein expression were lower in the HIF-2 RNAi group, the difference was significant (P<0.01). No significant difference between empty vector group and control group was observed. The cell growth in the HIF-2 RNAi group become slower. Compared with control group, the growth of tumor was slower in the RNAi group in the nude mice (P<0.01). CONCLUSION: HIF-2 RNAi inhibits the expression of 786-0 and cell growth, and slows the growth of tumor in the nude mice. The result provides new application for biological therapy in the renal cell cancer.