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- W2387717010 abstract "Objective To construct the lentivirus shRNA vector targeting Akt2.Cell clones inhibiting Akt2 expression were obtained,and it offered information for the discussion of Akt2.Method Sequences for targeting the Akt2 gene was selected.The double strand shRNA oligo was ligated to pGCsil-GFP lentivirus vector.The construct was verified by sequencing.Then a lentiviral expression vector,pHelper1.0,and p-Helper 2.0 plasmid vectors were cotrasnfected into 293T cells by Lipofectamin 2000.The viral particles were collected and infected A549 cells.After selection of GFP positive cells by FACS,mRNA and protein expression levels of Akt2 were determined by qPCR and western blot,respectively.Results The sequencing results confirmed that pGCsil-shAkt2-GFP vector was successfully constructed.After virus packaging,the concentrated virus titer was 3E+9 TU/ml.shAkt2 stable transfected cells were purified by FACS with GFP marker.The ratio of selection is 90.1%.Efficacy of Akt2 knockdown was determined by qRT-PCR and western blot.Our data showed that Akt2 mRNA level of shAkt2 group was significantly decreased by 63.39% compared with control groups,and protein expression level of Akt2 was inhibited by 91.56%.Stable knockdown of Akt2 in A549 cells was successfully established.Conclusions shAkt2 can significantly inhibit the Akt2 expression levels in A549 cell.It establishes the foundation for the study on effects of Akt2 on NSCLC." @default.
- W2387717010 created "2016-06-24" @default.
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- W2387717010 date "2011-01-01" @default.
- W2387717010 modified "2023-09-27" @default.
- W2387717010 title "Target-specific inhibition of Akt2 in non-small cell lung cancer A549 cell line mediated by lentivirus vector" @default.
- W2387717010 hasPublicationYear "2011" @default.
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