RNA干扰缺氧诱导因子2对肾癌786-0细胞的影响

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

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-2RNAi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。

AIM： To study the effect of RNA interference on hypoxia- inducible factor-2 （H IF -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.