Effect of transfected bone morphogenetic protein-2 gene and insulin-like growth factor gene on gene expression of Wistar rat mesenchymal stem cells

Abstract

Objective To investigate whether bone morphogenetic protein-2 (BMP-2) and insulin-like growth factor-1 ( IGF-1 ) in high glucose can be transfected into Wistar rat mesenchymal stem cells,and the gene expression after transfection. Methods Adenovirus-mediated BMP-2 and IGF-1 genes were transfected into the Wistar rat mesenchymal stem cells. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the gene replication and expression. Results After the transfection of adenovirus-mediated BMP-2 and(or) IGF-1 gene into the cells in high glucose, all of bone marrow stromal cells survived, bone marrow stromal cells could express the target gene product with the peak of BMP-2 gene product expression at in 72nd h after transfection. The grey degree and gene product concentration of BMP-2 in BMP-2 group were ( 1.86 ± 0. 06) and (58.60 ±1.61) μg/L, and those in BMP-2 ± IGF-1 group (2.37 ±0.34) and (67.61 ±1.91) μg/L,respectively (all P ＜ 0. 05 ). The peak of IGF-1 gene expression occurred at 48th h after transfection. The grey degree and gene product concentration of IGF-1 in IGF-1 group were ( 1.48 ± 0. 03 ) and ( 65.17 ±3.64) μg/L, and those in BMP-2 + IGF-1 group were (1.93 ±0. 17) and (73. 94 ±4. 30) μg/L, respectively ( all P ＜0. 05). Conclusion Adenovirus-mediated BMP-2 and IGF-1 gene could be transfected the Wistar rat bone marrow stromal cells in high glucose. The gene products after transfection were increased significantly. There was a synergistic effect on gene expression between BMP-2 and IGF-1. Key words: BMP-2; IGF-1; High glucose; Bone marrow mesenchymal stem cells