RNA interference by expression of short hairpin RNAs suppresses bcl-xL gene expression in nasopharyngeal carcinoma cells
Abstract
Aim:To evaluate a new plasmid mediated RNA interference (RNAi) system and
investigate whether knock-down of bcl-xL by short hairpin RNA (shRNA) can
induce apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-2Z in
vitro.
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 expressing 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 recombinant
plasmid efficiently suppressed bcl-xL gene expression and induced apoptosis
of NPC cells in vitro.
Conclusion: The recombinant plasmid can sufficiently
mediate RNAi in CNE-2Z cells, and knock-down of the bcl-xL expression by shRNA
significantly induced apoptosis in CNE-2Z cells. The results suggest this new
system, mediated RNAi can be used as a tool for the study of gene function and
gene therapy.
Keywords:
investigate whether knock-down of bcl-xL by short hairpin RNA (shRNA) can
induce apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-2Z in
vitro.
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 expressing 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 recombinant
plasmid efficiently suppressed bcl-xL gene expression and induced apoptosis
of NPC cells in vitro.
Conclusion: The recombinant plasmid can sufficiently
mediate RNAi in CNE-2Z cells, and knock-down of the bcl-xL expression by shRNA
significantly induced apoptosis in CNE-2Z cells. The results suggest this new
system, mediated RNAi can be used as a tool for the study of gene function and
gene therapy.