Construction of green fluorescent protein-bone mesenchymal stem cell-TNF-related apoptosis-inducing ligand vector and detection of its biology characteristics

Objective To explore the gene expression in bone mesenchymal stem cells (BMSCs) carried TNF-related apoptosis-inducing ligand (TRAIL) vector.Methods GFP-BMSCs were used in our study and divided into three groups:the experimental group,being transfected TRAIL gene to BMSCs,the empty vector group,being transfected with empty vector,and the blank control group,being transfected with lipidosome.Liposomes method was used to transfect TRAIL into GFP-BMSCs in the experimental group.The mRNA expression of TRAIL gene was detected by real time-PCR; Western blotting and immunofluorescence were used to detect the protein expression of TRAIL gene; the proliferation activity of BMSC cells was detected by MTT method.Results GFP-BMSC-TRAIL vectors were successfully constructed.Immunofluorescence indicated that,24 and 48 h after TRAIL transfection,TRAIL protein expression was noted in the cytoplasm and membrane,and that at 24 h was obviously stronger than that at 48 h; no expression was noted in the empty vector group and blank control group.Real time PCR and Western blotting indicated TRAIL high expression in the experimental group,while not in the blank control and empty vector group.MTT showed no significant changes of BMSCs activity after TRAIL transfection.Conclusion BMSC-TRAIL vectors can stably express the target gene,indicating that BMSCs may be an effective vector for gene therapy. Key words: TNF-related apoptosis-inducing ligand;  Bone mesenchymal stem cell;  Vector