study on transduction of il 24 gene in human bone marrow mesenchymal stem cells by lentiviral vector

• DOI: 10.3779/j.issn.1009-3419.2013.01.02
• Publication Date: 2013-01-01
• Authors: Weiqiang Li; Qiugan Qi; Qinghua Zhou; Yin Li; Fei Chen; Jianjun Qin
• Keywords: Interleukins; Genetic Vectors; Green Fluorescent Proteins; Lentivirus; EGFP; Neoplasms. Tumors. Oncology. Including cancer and carcinogens; Bone Marrow Cells; Mesenchymal Stem Cells; Genetic Therapy; Viral Load; hBMSCs; Gene Expression Regulation; Genes, Reporter; Transduction, Genetic; IL-24; Humans; Gene transduction; Lentiviral vector; RC254-282

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.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.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.Lentiviral vector carrying recombinant IL-24 gene can effectively transduce hBMSCs and express IL-24 protein.