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.
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Project supported by grants from the Natural Science Foundation of Guangdong Province (No 31962), the Special Funds for Major Research Program of Guangdong Province (No ZKM04205S), and the Special Funds for Major Subject Program of Guang dong Province (No 9906).
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Liu, F., He, Cw., Zhang, Yf. et al. RNA interference by expression of short hairpin RNAs suppresses bcl-xL gene expression in nasopharyngeal carcinoma cells. Acta Pharmacol Sin 26, 228–234 (2005). https://doi.org/10.1111/j.1745-7254.2005.00529.x
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DOI: https://doi.org/10.1111/j.1745-7254.2005.00529.x
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