%0 Journal Article
%T Study on Transduction of IL-24 Gene in Human Bone Marrow Mesenchymal Stem Cells by Lentiviral Vector
%A Qiugan QI
%A Qinghua ZHOU
%A Yin LI
%A Weiqiang LI
%J Chinese Journal of Lung Cancer
%D 2013
%I Chinese Anti-Cancer Association; Chinese Antituberculosis Association
%R 10.3779/j.issn.1009-3419.2013.01.02
%X 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¡Á107 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 ¦Ìg/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.
%K hBMSCs
%K IL-24
%K EGFP
%K Gene transduction
%K Lentiviral vector
%U http://dx.doi.org/10.3779/j.issn.1009-3419.2013.01.02