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- W2388551423 abstract "Objective To explore inhibitory effect of RNA interference with eukaryotic expression plasmid containing shRNA on expression of mTOR gene in human colon cell line LoVo.Methods Specific DNA sequences encoding shRNA were synthesized and inserted into plasmid pGenesil-1.Then these recombinant plasmids were transfected into LoVo cells with LipofectamineTM 2000 and the transfected cells were selected for 4 weeks by using G418.There were three groups in this study: group A the normal cultured LoVo cells untransfected;group B LoVo cells transfected with plasmid pGenesil-1 as negative control;group C LoVo cells transfected with pGenesil-1-mTOR plasmid vectors.Moreover,the mRNA expression of mTOR was assessed by RT-PCR,and the protein expression of mTOR gene was detected via Western blot analysis.Results Agarose gel electrophoresis of plasmids incised with two enzymes showed that a 410 bp or 350 bp DNA segment was incised by enzymatic digestion in the recombinant plasmid pGenesil-1-mTOR or empty plasmid pGenesil-1 respectively.The green fluorescence was observed in the transfected cells under inversion fluorescence microscope.The mRNA expression of mTOR gene in group C was significantly lower than that in group A(P0.05),which inhibition rate was 73.0%.The protein expression of mTOR in group C was significantly lower than that in group A(P0.05),which inhibition rate was 72.5%.Conclusion The expression of mTOR gene in colon cancer LoVo cells could be suppressed efficiently by RNA interference with eukaryotic expression plasmid containing shRNA." @default.
- W2388551423 created "2016-06-24" @default.
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- W2388551423 date "2008-01-01" @default.
- W2388551423 modified "2023-09-26" @default.
- W2388551423 title "Inhibition Effect of RNA Interference with Eukaryotic Expression Plasmids Containing shRNA on mTOR Gene Expression in Human Colon Cancer LoVo Cell Line" @default.
- W2388551423 hasPublicationYear "2008" @default.
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