Open AccessRNA interference by expression of short hairpin RNAs suppresses bcl ‐xL gene expression in nasopharyngeal carcinoma cells 1
Author(s) -
LIU Fang,
HE Chengwei,
ZHANG Yuefei,
ZHOU Keyuan
Publication year - 2005
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2005.00529.x
Subject(s) - small hairpin rna , rna interference , microbiology and biotechnology , transfection , plasmid , biology , gene expression , bcl xl , gene , recombinant dna , rna , small interfering rna , apoptosis , chemistry , programmed cell death , genetics
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.