title: [Recombinant adenovirus of microRNA-193b inhibits proliferation of K562 cells].
abstract: OBJECTIVE
  To construct a recombinant adenovirus vector containing human pre-miR-193b and investigate its effect on the proliferation of chronic myelocytic leukemia K562 cells.
  
  
  METHODS
  The cDNA of pre-miR-193b was obtained by chemical synthesis and inserted into the adenoviral shuttle vector pAdTrack-CMV. The recombinant shuttle plasmid was linearized by Pme I and transformed into AdEasier cells for homologous recombination in E.coli BJ5183 with the adenoviral backbone plasmid pAdEasy-1. The recombinant adenoviral plasmid was linearized by Pac I and then used for transfecting HEK293 cells. After package and amplification in HEK293 cells, the virus titer was determined by serial dilution assay. The expression level of miR-193b in K562 cells was detected by real-time quantitative PCR. The cell proliferation was observed by MTT assay.
  
  
  RESULTS
  The recombinant plasmid named pAd-miR-193b was confirmed by restriction enzyme analysis and sequencing. The recombinant adenovirus could infect K562 cells efficiently. Compared with control group, real-time quantitative PCR revealed that the expression of miR-193b was significantly up-regulated in K562 cells infected with pAd-pre-miR-193b, and the cell proliferation was suppressed significantly in K562 cells with up-regulated miR-193b expression.
  
  
  CONCLUSIONS
  The over-expression of miR-193b may suppress the proliferation of K562 cells.
authors: Department of Immunology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.
authors: Yan
authors: Y
authors: Shu
authors: Department of Immunology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.
authors: Jieyu
authors: J
authors: Qi
authors: Department of Immunology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.
authors: Canwei
authors: C
authors: Wang
authors: Department of Immunology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.
authors: Shujing
authors: S
authors: Yu
authors: Department of Immunology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.
authors: Kun
authors: K
authors: Tao
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D015536:Down-Regulation; D057809:HEK293 Cells; D006801:Humans; D020014:K562 Cells; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology*; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism
chemical_list: C546818:MIRN193 microRNA, human; D035683:MicroRNAs