Adeno-associated virus-mediated BMP-7 and SOX9 in vitro co-transfection of human degenerative intervertebral disc cells

转染 硫氧化物9 骨形态发生蛋白2 椎间盘 骨形态发生蛋白7 体外 分子生物学 腺相关病毒 骨形态发生蛋白 细胞生物学 化学 生物 基因表达 重组DNA 解剖 基因 生物化学 载体(分子生物学)
作者
Xingjie Ren,Zezheng Diao,Yongming Xi,Zonghua Qi,Shan Ren,Y J Liu,Deling Yang,X Zhang,Si-Ming Yuan
出处
期刊:Genetics and Molecular Research [Research Foundation of Ribeirão Preto]
卷期号:14 (2): 3736-3744 被引量:13
标识
DOI:10.4238/2015.april.22.1
摘要

Bone morphogenetic protein-7 (BMP-7) and SOX9 are important transcription factors in chondrogenesis. In this study, we examined the biological function of the adeno-associated virus (AAV)-mediated BMP-7 and SOX9 double gene in vitro co-transfection of nucleus pulposus cells of human degenerative intervertebral disc. Human intervertebral disc nucleus pulposus cells were cultured in vitro and subcultured for 5 generations. Using rAAV-BMP-7 and rAAV-SOX9 AAV2-type AAV viruses, the cells were divided into 4 groups: blank normal, BMP-7 transfection, SOX9 transfection, and BMP-7 and SOX9 co-transfection. After 48 h, expression of type II collagen and its mRNA in nucleus pulposus cells was determined. The expression of type II collagen in BMP-7 transfection, SOX9 transfection, and co-transfection groups was up-regulated to varying degrees compared to the blank control group. The type II collagen mRNA level expression in the co-transfection group was significantly higher than in other groups (P < 0.05). AAV-mediated BMP-7 and SOX9 in vitro co-transfection can promote the synthesis of type II collagen in nucleus pulposus cells in the human degenerative intervertebral disc. Double-gene therapy has a synergistic effect in the treatment of intervertebral disc degeneration.

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