title: [Expression of a chimeric gene containing poly-arginine as the protein transduction domain and its killing activity against HER2 positive gastric cancer SGC-7901 cells].
abstract: OBJECTIVE
  To construct a eukaryotic expression vector for chimeric gene containing poly-arginine as the protein transduction domain(PTD) and transiently transfect this vector into HER2 positive SGC-7901 cells and HER2 negative HeLa cells to examine its effect on cell growth.
  
  
  METHODS
  PCR amplication was used to obtain the gene of active form caspase-3 fused with nonaarginine, and then fusion gene was cloned into eukaryotic expression vector containing e23sFv DNA fragment. After this chimeric gene transfected into SGC-7901 cells and HeLa cells by Lipofectamine 2000™; reagent, indirect immunofluorescence and cell counting were used to examine the expression in these two cells and the effect on cell growth.
  
  
  RESULTS
  The eukaryotic expression vector, named pCMV-e23sFv-R₉;-casp3, encoding e23sFv/caspase-3 containing nonaarginine as the PTD was successfully constructed. e23sFv- R₉;-casp3 protein was expressed in a secretary manner in both SGC-7901 cells and HeLa cells. Transfected SGC-7901 cells were found obvious growth inhibitory, morphology change and condensed nucleus, whereas neither growth inhibitory nor apparent morphology change was detected in transfected HeLa cells.
  
  
  CONCLUSIONS
  Of the secretary expressed chimeric protein, the antibody moiety against HER2 can mediate targeted recognition, the nonaarginine translocating peptide can promote activation and translocation of the effector molecule, and the active caspase-3 can effectively induce cell killing.
authors.affiliation: Department of Immunology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Zhi-ning
authors.initials: ZN
authors.lastname: Zhao
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.forename: Jun-lin
authors.initials: JL
authors.lastname: Ren
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.forename: Wei-hong
authors.initials: WH
authors.lastname: Wen
authors.forename: Lin-tao
authors.initials: LT
authors.lastname: Jia
authors.forename: An-gang
authors.initials: AG
authors.lastname: Yang
mesh_terms: D017209:Apoptosis; D053148:Caspase 3 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D010455:Peptides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013274:Stomach Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology*
chemical_list: D053148:Caspase 3; D010455:Peptides; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; C015462:polyarginine; C508053:ERBB2 protein, human