title: [Study on transduction of IL-24 gene in human bone marrow mesenchymal stem cells by lentiviral vector].
abstract: BACKGROUND
  Up to know, no any study on using human bone marrow mesenchymal stem cells (hBMSCs) as cells carrier of tumor suppressor gene (IL-24) was reported. The aim of this study is to study the efficiency of transduction of hBMSCs by constructing the lentiviral vector in co-expressing enhanced green fluorescent protein (EGFP) gene and human IL-24 gene, and to lay a foundation for gene therapy of tumor in the future.
  
  
  METHODS
  The lentivector which contain IL-24 and EGFP constructed by recombinant DNA technology were co-transfected to 293FT cells with ViraPowerTM Lentiviral Packaging Mix. The recombinant lentivirus infected with hBMSCs were selected and purified by puromycin. Expression of IL-24 mRNA and IL-24 protein levels were detected by real-time quantitative PCR (qPCR) and ELISA.
  
  
  RESULTS
  The recombinant lentiviral vector of co-expressing IL-24 gene and EGFP gene were successfully constructed by multisite Gateway technology, virus can be packaged, purified and concentrated successfully, and the virus titer was 7.25×10⁷ PFU/mL. The efficiency of recombinant lentivirus to transduce hBMSCs can reach 100% after selection. The result of qPCR showed that the level of IL-24 mRNA expression in transduced group was significantly higher than that in non-transduced group (P<0.05); ELISA detection confirmed that IL-24 protein expression of transduced group was positive in supernatant and the concentration of IL-24 protein is 40 μg/L, while the non-transduced group was negative.
  
  
  CONCLUSIONS
  Lentiviral vector carrying recombinant IL-24 gene can effectively transduce hBMSCs and express IL-24 protein.
authors: Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors: Qiugan
authors: Q
authors: Qi
authors: Qinghua
authors: Q
authors: Zhou
authors: Yin
authors: Y
authors: Li
authors: Weiqiang
authors: W
authors: Li
authors: Fei
authors: F
authors: Chen
authors: Jianjun
authors: J
authors: Qin
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology*; D005786:Gene Expression Regulation / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007378:Interleukins / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics* / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods*; D019562:Viral Load; D000099180:Interleukin-24
chemical_list: D049452:Green Fluorescent Proteins; D007378:Interleukins; D000099180:Interleukin-24; C485184:enhanced green fluorescent protein