title: [Construction and application of a recombinant plasmid inducing cell death in hepatocytes after transfection].
abstract: OBJECTIVE
  The mechanisms of mesenchymal stromal cells (MSCs)-mediated treatment of liver damage have been unclear. Two major mechanisms, which involve paracrine effects and/or direct trans-differentiation, have been proposed. To clarify which mechanism is more important, we planned to construct a recombinant plasmid expressing green fluorescent protein (GFP) driven by the cytomegalovirus (CMV) promoter and Pseudomonas aeruginosa exotoxin 40 (PE40) driven by the albumin (alb) promoter, which can induce cell death as soon as MSCs differentiate into hepatocytes.
  
  
  METHODS
  To construct the recombinant eukaryotic expression vector pFlag-CMV-GFP-TM-albp-PE40, GFP, transmembrane domain of DLL1, alb promoter and PE40 were obtained by PCR and were inserted into pFlag-CMV-1. The expression of GFP was observed under a fluorescence microscope and the killing effect on hepatocytes was analyzed by flow cytometry.
  
  
  RESULTS
  The recombinant plasmid inducing cell death in hepatocytes was successfully constructed, suggesting that the plasmid could be employed to study the mechanism of MSCs-mediated treatment on liver damage.
  
  
  CONCLUSIONS
  This study might provide a promising tool for revealing the mechanism of MSCs-mediated treatment on liver diseases.
authors: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors: Chunchen
authors: C
authors: Gao
authors: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors: Junlong
authors: J
authors: Zhao
authors: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors: Jun
authors: J
authors: Han
authors: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors: Dongdong
authors: D
authors: Li
authors: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors: Gaoyang
authors: G
authors: Zhao
authors: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors: Hua
authors: H
authors: Han
authors: Department of Medical Genetics and Developmental Biology, School of Basic Medicine, Fourth Military Medical University, Xi'an 710032, China.
authors: Hongyan
authors: H
authors: Qin
mesh_terms: D016923:Cell Death; D002454:Cell Differentiation; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins