LIM mineralization protein expression in LIM mineralization protein-1 gene-transfected bone marrow mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:In vitro experiments suggest that LIM mineralization protein-1 (LMP-1) gene can increase the expression of LMP-1 protein in osteoporotic bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the LMP-1 expression in osteoporotic bone marrow mesenchymal stem cells transfected with RV-LMP-1-GFP in vitro. METHODS:Twelve SD female rats were selected and subjected to bilateral ovariectomy for establishment of osteoporosis models. After 2 months of feeding, bilateral femurs, tibiae, and humeri of rats were taken to isolate and culture bone marrow mesenchymal stem cells. Passage 3 cells were taken and randomly divided into ovariectomized group and LMP-1 transfection group. Another six rats only underwent removal of the same amount of fat tissue around the ovary, and passage 3 cells which were harvested as those in the former two groups served as sham group. RT-PCR and western blot assay were performed to determine the expression of LMP-1 protein and mRNA. RESULTS AND CONCLUSION:Under an inverted fluorescence microscope, transfected bone marrow mesenchymal stem cells from osteoporotic rats showed green fluorescent expression. The three groups al could express LMP-1 at the protein and mRNA levels. The expression levels of LMP-1 protein and mRNA in the LMP-1 transfection group were significantly higher than those in the ovariectomized group and sham group (P0.05). The successful expression of the RV-LMP-1-GFP gene in osteoporotic SD rat bone marrow mesenchymal stem cells was realized at the protein and mRNA levels;moreover, the expression of LMP-1 was increased dramatical y. These findings indicate that LMP-1 gene is successful y transferred into osteoporotic SD rat bone marrow mesenchymal stem cells, and significantly elevates the expression of LMP-1 mRNA and protein.