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Abstract To construct the antisense nucleic acid eukaryotic expression vector of human cy-clooxygenase-2(COX-2) ,and to explore its role in inhibiting the expression of COX-2 in human bladder cancer T24 cells .[Methods] PCR method was used to amplify cDNA from COX-2 plasmid .The COX-2 cDNA was cloned into the sites between T vectors EcoR 1 and Xba I through restrictive endonuclease ,and then inversely inserted into pcDNA3 .1 to obtain the recombinant plasmid .The recombinant plasmid was transfected into bladder cancer T24 cells mediated by liposome .After the scanning by G418 ,flow cytometry ,indirect immuno-fluorescence staining and RT-PCR were performed .[Results] Nucleic acid and restrictive endonuclease diges-tion confirmed that COX-2-antisense nucleic acid eukaryotic expression plasmid pcDNA 3 .1/COX-2 was cor-rect .Identification showed that the expression of COX-2 in T24 cells transfected with pcDNA3/COX-2 was in-hibited obviously .[Conclusion] Antisense nucleic acid eukaryotic expression vector of COX-2 is constructed successfully .Bladder cancer cell line T24-AS with persistent downregulation of COX-2 expression is obtained .
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2014, Vol. 31 
