Construction and Identification of Vector Expressing RNA Interference Aimed at Human Cyclin D_1 Gene

[Purpose] To construct the eukaryotic expression vector for RNA interference human cyclin D1 gene and detect its interference effect in human ovarian cancer cell line(HO-8910). [Methods] Four target gene segments were synthesized and cloned into pSUPER vector respectively to construct four recombinant eukaryotic expression vectors pSUPER-C1～4 based on targeted cyclin D1 gene. The four recombinant vectors were identified by enzyme digestion analysis and DNA sequencing. Then HO-8910 cells were transfected with pSUPER-C1～4 and subjected to G418 selection. In G418-resistant cells, the interference effect was detected by RT-PCR and effective sequence-specific siRNA was screened out. [Results] Enzyme digestion analysis and DNA sequencing showed that the target segments were cloned into pSUPER vector. The four recombinant vectors inhibited transcription of cyclin D1 gene with different extent and pSUPER-C2 had most intensive interference effect. [Conclusion] The sequence-specific siRNA which could effectively interfer cyclin D1 gene expression has been screened out. The transcription and expression of cyclin D1 gene are inhibited effectively by the constructed RNAi eukaryotic expression vectors in the ovarian cancer cells.