title: [Construction of the expression vector for short-hairpin RNA-mediated Akt gene silencing in Lovo cells].
abstract: OBJECTIVE
  To construct a eukaryotic expression vector of short-hairpin RNA (shRNA) targeting human Akt gene and assess the effect of Akt gene silencing on the growth of colon cancer Lovo cells.
  
  
  METHODS
  Two shRNAs targeting human Akt gene were cloned into pENTRTM/U6 plasmid to obtain the entry clones, and the positive clones were verified by sequencing. After recombination of the pENTRTM/U6 entry constructs and Plenti6/Block-iT DEST vector, the positive clones were confirmed by sequencing. Lovo cells were transfected by the entry vector and DEST Vector, and RT-PCR and Western blotting were performed to detect the interference of Akt gene expressions.
  
  
  RESULTS
  The pENTRTM/U6 entry clones carrying Akt shRNA and pLenti6/DEST-pENTRTM/U6-Akt shRNA were successfully constructed. Both of the vectors were transfected into Lovo cells and resulted in obvious knockdown of the mRNA and protein expressions of Akt.
  
  
  CONCLUSIONS
  The Akt siRNA expression vector constructed can significantly inhibit Akt gene expression in Lovo cells, which facilitates further studies of Akt function and tumor gene therapy.
authors: Clinical Laboratory, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, China. huangxz020@163.com
authors: Xian-zhang
authors: XZ
authors: Huang
authors: Xue-qin
authors: XQ
authors: Zhao
authors: Man
authors: M
authors: Li
authors: Min
authors: M
authors: He
authors: Jun-hua
authors: JH
authors: Zhunag
mesh_terms: D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000473:pathology*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051057:Proto-Oncogene Proteins c-akt / Q000096:biosynthesis / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: D034741:RNA, Small Interfering; D051057:Proto-Oncogene Proteins c-akt