Study on Transduction of IL-24 Gene in Human Bone Marrow Mesenchymal Stem Cells by Lentiviral Vector

Abstract

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

Keywords

• hBMSCs
• IL-24
• EGFP
• Gene transduction
• Lentiviral vector