关键词: RNA干扰, 肿瘤

Abstract: Objective To construct the expression vectors of pSilencer3.1 -hif-1α and identify the inhibition of hif-1α in Hela299 cells after transfection with the combinative plasmids. Methods The interfering sequences of hif-1α were designed according to the sequence of hif-1α of GenBank. Three recombinant plasmids of pSilencer3.1-hif-1α were constructed by DNA ligase. Trizol reagent was used to extract the whole RNA of cells and RT-PCR assay was applied to detect the expression of hif-1α mRNA . Lysis assay was used to extract the protein from cells and Western blotting was adopted to observe the expression of HIF-1α protein. Results The vectors were identified to be right after sequencing. The mRNA level was decreased 24 h after transfection with three vectors of pSilencer-hif-1α, and the ratios of hif-1α /GAPDH in control, group transfecting with pSilencer-sihif-1α-1, group transfecting with pSilencer-sihif-1α-2, group transfecting with pSilencer-sihif-1α-3 were 0.55, 0.13, 0.33, and 0.08, respectively(P<0.05, P<0.05, P<0.01). The protein expression levels of HIF-1α 48 h after transfection with three vectors of pSilencer-hif-1αwere decreased significantly. Conclusion Effective pSilencer-hif-1α plasmids are constructed successfully and they could interfere with the expression of hif-1α in Hela299 cells.

Key words: RNA interference, neoplasms