KDR siRNA expression vector and identification of.docVIP

 PAGE \* MERGEFORMAT 5 KDR siRNA expression vector and identification of Author: Miao Ying industry, Liu Yun, Yi Jing, Su rights, Chen Jianjun, Hao Ke [Keywords:] vascular endothelial growth factor receptor 2 Construction and identification of siRNA expression vector targeting KDR [Abstract] AIM: To construct pSilencerTM3.1H1 small inferfering RNA (siRNA) expression vector targeting human KDR gene and to observe its silencing effect in the PC3 prostate cancer cell line. METHODS: The designed oligos targeting KDR were cloned into the pSilencerTM3.1H1 neo vector, which was lineated after BamH Ⅰ and Hind Ⅲ digestion. The recombimant vectors were confirmed by enzyme digestion analysis and DNA sequencing.The recombimant vectors of pSilencer 3.1KDR1, pSilencer 3.1KDR2 and pSilencer 3.1KDR3 were transfected by liposomemediated transfection into the PC3 prostate cancer cell line with high metastasis potential. At 72 h after transfection, the expression of KDR at the levels of mRNA and protein was detected by RTPCR and Western blotting. RESULTS: The pSilencerTM3.1H1 siRNA expression vectors were constructed and confirmed after the enzyme digestion analysis and the DNA sequencing. The siRNA expression vectors of pSilencer 3.1KDR1, pSilencer 3.1KDR2 and pSilencer 3.1KDR3, were successfully constructed.pSilencer 3.1KDR3 was most effective in the 3 which downregulated 55% mRNA and protein of KDR at 72 h after transfection. CONCLUSION: pSilencerTM3.1H1 siRNA expression vectors targeting KDR were successfully constructed. The expression of KDR gene was inhibited effectively in PC3 cells transfacted by pSilencer 3.1KDR3, which laid a basis for its application in the treatment of prostate cancer. [Keywords] vascular endothelial growth factor receptor2; RNA, small interfering; PC3 cell line [Abstract] Objective: To construct targeting vascular endothelial growth factor receptor Ⅱ (VEGFR2), also known as functional area receptor kinase (KDR) in pSilencerTM3.1 siRNA (smal