title: Preparation and characterization of nano-liposome-mediated FL gene in the Lovo cells.
abstract: OBJECTIVE
  The purpose of the present work was to formulate and evaluate cationic nano-liposomes as novel nonviral gene delivery for colon cancer treatment.
  
  
  METHODS
  Recombinant pEGFP-c1-Fms-like tyrosine kinase receptor 3 ligand (FL) plasmids containing human FL gene and green fluorescent protein (GFP) reporter genes were constructed. FL and GFP Gene-carrying cationic nano-liposomes were prepared based on the electrostatic adherence principle and then transfected into Lovo cells. The morphology, particle size, and zeta potential of gene-carrying cationic nano-liposomes were observed using an electron microscope. GFP expression was observed by fluorescence microscopy to assay the transfection efficiency. The cytotoxicity of FL/nano-liposomes was evaluated by the MTT method.
  
  
  RESULTS
  Recombinant plasmids pEGFP-c1-FL are successfully constructed using gene cloning methods and confirmed by restriction enzyme digestion and sequencing. The cationic nano-liposomes carrying pEGFP-cl-FL were observed by an electron micrograph and showed uniform spherical or elliptical shapes and many pores. The fluorescence microscopy images of gene-carrying cationic nano-liposomes showed good expression of GFP in pEGFP and pEGFP-cl-FL groups. The MTT assay of cell death indicated a significantly higher level of cell death between the FL group and the control group at 24, 48, and 96 hours after transplantation.
  
  
  CONCLUSIONS
  Cationic nano-liposomes show safe and high-performance transfection as gene carriers. Gene therapy has significant implications for colon cancer treatment in future.
authors.affiliation: Department of General Surgery, Qilu Hospital of Shandong University , Jinan, China.
authors.forename: Nian-Feng
authors.initials: NF
authors.lastname: Sun
authors.forename: Ai-Ling
authors.initials: AL
authors.lastname: Tian
authors.forename: Yong-Xin
authors.initials: YX
authors.lastname: Hua
authors.forename: San-Yuan
authors.initials: SY
authors.lastname: Hu
authors.forename: An-Bin
authors.initials: AB
authors.lastname: Hu
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics / Q000628:therapy*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry; D008297:Male; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D014162:Transfection / Q000379:methods*
chemical_list: D008081:Liposomes; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C084881:flt3 ligand protein; D049452:Green Fluorescent Proteins