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Aim:To evaluate a new plasmid mediated RNA interference(RNAi)system andinvestigate whether knock-down of bcl-xL by short hairpin RNA(shRNA)caninduce apoptosis of human nasopharyngeal carcinoma(NPC)cell line CNE-2Z invitro.Methods:The plasmid containing mU6 promoter was subcloned to yield thepmU6 plasmid,recombinant plasmid expressing shRNA targeting bcl-xL gene wasdesigned and constructed,and were co-transfected cells with green fluorescenceprotein expressing plasmid.Flow cytometry was used to evaluate transfectionefficiency,and RT-PCR and Western blot were applied to analyze bcl-xL mRNAand protein levels,respectively.Results:The shRNA expressed by the recombi-nant plasmid efficiently suppressed bcl-xL gene expression and induced apoptosisof NPC cells in vitro.Conclusion:The recombinant plasmid can sufficientlymediate RNAi in CNE-2Z cells,and knock-down of the bcl-xL expression by shRNAsignificantly induced apoptosis in CNE-2Z cells.The results suggest this newsystem,mediated RNAi can be used as a tool for the study of gene function andgene therapy.
Aim: To evaluate a new plasmid mediated RNA interference (RNAi) system and investigated whether knock-down of bcl-xL by short hairpin RNA (shRNA) caninduce apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-2Z invitro.Methods: The plasmid containing mU6 promoter was subcloned to yield the pmU6 plasmid, recombinant plasmid expressing shRNA targeting bcl-xL gene was designed and constructed, and were co-transfected cells with green fluorescence protein clear plasmid. Flow cytometry was used to evaluate transfection efficiency, and RT-PCR and Western blot were applied to analyze bcl-xL mRNA and protein levels, respectively. Results: The shRNA expressed by the recombi-ntry plasmid efficiently suppressed bcl-xL gene expression and induced apoptosis of NPC cells in vitro. Confluence: The recombinant plasmid can be used as RNAi in CNE- 2Z cells, and knock-down of the bcl-xL expression by shRNAsificantly induced apoptosis in CNE-2Z cells. The results suggest this newsystem, mediated RNAi can be used as a tool for the study of gene function and gene therapy.