Zhonghua yi xue za zhi · 2007
[Inhibition of the aquaporin-1 gene expression by RNA interference: experiment with cultured rat pleural mesothelial cells].
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Zhang Wei, Xie Can-mao, Li Zhi-ping, Zhu Zhi-wen, Yan Ying-shuo, DU Hong-chun
Аннотация
OBJECTIVE: To investigate the influence of RNA interference (RNAi) on the expression of aquaporin-1 (AQP1) gene and to investigate the feasibility of gene therapy for pleural effusion.
METHODS: Two recombinant plasmids with shRNAs targeting the AQP1 gene, AQP1-1-pGenesil and AQP1-2-pGenesil-1 were constructed. Pleural mesothelial cells were obtained from rats, cultured, and randomly divided into 5 groups: blank control group, Lipofectamine 2000 control group, HK negative control group, AQP1-1-pGenesil-1 transfected group, and AQP1-2-pGenesil-1 transfected group. RT-PCR and Western blotting were used to detect the mRNA and protein expression of AQP1.
RESULTS: The mRNA expression levels of aquaporin-1 of the AQP1-1-pGenesil-1 and AQP1-2-pGenesil-1 transfected groups were inhibited by 83.5% and 90.9% respectively, and the protein expression levels of the AQP1-1-pGenesil-1 and AQP1-2-pGenesil-1 transfected groups were inhibited by 41.2% and 67.6% respectively.
CONCLUSION: RNAi can successfully inhibit the expression of AQP1 and has the feature of sequence correlation of shRNA in the cultured rat pleural mesothelial cells. It may be used as a potential new approach for gene therapy of pleural effusion.
METHODS: Two recombinant plasmids with shRNAs targeting the AQP1 gene, AQP1-1-pGenesil and AQP1-2-pGenesil-1 were constructed. Pleural mesothelial cells were obtained from rats, cultured, and randomly divided into 5 groups: blank control group, Lipofectamine 2000 control group, HK negative control group, AQP1-1-pGenesil-1 transfected group, and AQP1-2-pGenesil-1 transfected group. RT-PCR and Western blotting were used to detect the mRNA and protein expression of AQP1.
RESULTS: The mRNA expression levels of aquaporin-1 of the AQP1-1-pGenesil-1 and AQP1-2-pGenesil-1 transfected groups were inhibited by 83.5% and 90.9% respectively, and the protein expression levels of the AQP1-1-pGenesil-1 and AQP1-2-pGenesil-1 transfected groups were inhibited by 41.2% and 67.6% respectively.
CONCLUSION: RNAi can successfully inhibit the expression of AQP1 and has the feature of sequence correlation of shRNA in the cultured rat pleural mesothelial cells. It may be used as a potential new approach for gene therapy of pleural effusion.