The aim of this study was to construct an RNA-interference plasmid (p-HIF-1 alpha RNAi) targeting the human HIF-1 alpha gene and assess its effects on HIF-1 alpha expression and its anti-tumour functions in vitro. p-HIF-1 alpha RNAi was constructed and confirmed by polymerase chain reaction (PCR) and DNA sequencing. Reverse transcriptase PCR (RT-PCR) and western blot were performed to detect HIF-1 alpha expression in HCT116 cells following transfection of p-HIF-1 alpha RNAi and p-control. The anti-tumour effects and mechanism of action of p-HIF-1 alpha RNAi in HCT116 cells were further investigated. p-HIF-1 alpha RNAi significantly inhibited HIF-1 alpha expression in the HCT116 cell line. p-HIF-1 alpha RNAi inhibited cell viability and reduced VEGF but not bFGF expression in the supernatant of HCT116 cells, down-regulated b-catenin and VEGF expression, and altered beta-catenin location in the HCT116 cell nucleus. The plasmid p-HIF-1 alpha RNAi can effectively and specifically inhibit HIF-1 alpha expression, inhibit cell proliferation, and alter the expression of key components in the Wnt/beta-catenin signaling pathway. Thus, p-HIF-1 alpha RNAi is a novel and extremely promising therapeutic inhibitor of HIF-1 alpha.
[1]Shanghai Univ Tradit Chinese Med, Putuo Hosp, Dept Gen Surg, Shanghai 200062, Peoples R China.
(8.0+极速模式)