title: [Construction of colorectal cancer cell line stably expressing mir-101 and identification of the target gene of mir-101].
abstract: OBJECTIVE
  To construct a colorectal cancer cell line stably expressing mir-101 and identify the target gene of mir-101.
  
  
  METHODS
  Quantitative real-time PCR was used to detect mir-101 expression in colorectal cancer cell lines. The recombined lentiviral vector GV209-mir101 or the empty lentiviral vector GV209 was transfected into human colorectal cancer cells SW620. The recombinant psiCHECK-2-Rac1 vector containing RAC1 3'UTR was constructed, and site-directed mutagenesis of RAC1 3'UTR was induced to construct the psiCHECK-2-Rac1-Mut vector. In HEK293A and SW480 cells co-transfected with mir-101 inhibitors or negative control (NC) and these recombined vectors, luciferase activities was examined with a dual-luciferase reporter assay.
  
  
  RESULTS
  SW620 cells transfected with GV209-mir101 lentivirus exhibited higher mir-101 expression level than cells transfected with GV209 lentivirus. Mir-101 inhibitors significantly increased the luciferase activities of RAC1 3'UTR. Overexpression of mir-101 increased the expression of RAC1 while inhibition of mir-101 suppressed RAC1 expression.
  
  
  CONCLUSIONS
  We have successfully constructed a SW620 cell line stably overexpressing mir-101. mir-101 can suppress RAC1 gene expression by targeting the specific sequence of RAC1 3'UTR.
authors: Department of Pathology, College of Basic Medical Sciences, Key Laboratory of Molecule Tumor Pathology of Guangdong Province, Guangzhou 510515, China. E-mail: liuyanruoshui@163.com.
authors: Yan
authors: Y
authors: Liu
authors: Yanxia
authors: Y
authors: Lu
authors: Min
authors: M
authors: Zhou
authors: Chao
authors: C
authors: Zhang
authors: Xuenong
authors: X
authors: Li
mesh_terms: D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection
chemical_list: C534072:MIRN101 microRNA, human; D035683:MicroRNAs