The construction and selection of HIF-2 RNA interference expression plasmid in the renal cell carcinoma

Aim:To construct eukaryotic expression vector carrying human HIF-2 RNAi and to study the effect of RNA interference on HIF-2 expression in the renal cell carcinoma.Methods:HIF-2 siRNA was designed.,two ssDNA oligonucleo-tides were synthesized.The annealed products were ligated into the pSilencer 2.1-U6 neo-vector.HIF-2 RNAi was inserted into RNA interference eukaryotic expression vector,For each sequence,All constructs were verified by standard DNA sequencing.the vector was transfected into renal cell cancer 786-0.Using Western Blot method to detect the protein expression of HIF-2 after transfection of RNAi vector using Lipofectamine.Results:HIF-2 RNAi eukaryotic expression vector was constructed.Compared with no transfect group and control group,the amount of HIF-2 protein expression are lower in the HIF-2 RNAi group,the amount of HIF-2 protein expression are lower in No 1、No 2 and No 3 cell groups.Their difference is significant(P0.05).There was no significance between no transfect cell groups and control cell groups in the the amount of HIF-2 protein expression(P0.05).Conclusion:The HIF-2 RNAi mammalian expression vector pSilencer 2.1-U6 neo was successfully constructed.HIF-2 RNAi can inhibit the protein expression of 786-0.Our study provide basis for biological therapy in the renal cell cancer.