PTEN基因过表达对BMSCs迁移的影响

Effect of over-expression of PTEN gene on migration of bone marrow mesenchymal stem cells

目的探讨PTEN基因过表达对大鼠骨髓来源间充质干细胞(BMSCs)迁移的影响。方法构建PTEN过表达的重组腺病毒载体,病毒扩增后,感染BMSCs,并设阴性对照组(Ad),感染48 h后,荧光定量PCR和western blot分别检测细胞中PTEN mRNA和蛋白水平的表达变化。同时,通过划痕实验检测PTEN基因过表达后,BMSCs迁移的变化。结果重组腺病毒载体经酶切鉴定证明构建正确。PTEN基因过表达后,BMSCs迁移受到明显抑制。结论体外实验证明,PTEN基因过表达后,BMSCs迁移受到明显抑制,为深入研究PTEN基因对BMSCs的迁移机制奠定一定的基础。

Objective To investigate the effect of over-expression of PTEN gene on the migration of rat bone marrow-derived mesenchymal stem cells.Methods The recombinant adenovirus vector with over-expression of PTEN was constructed and the adenovirus was amplified.After 48 hours of infection with BMSCs,the expression of PTEN mRNA and protein was detected by fluorescence quantitative PCR and western blot,respectively.At the same time,the scratch test was used to detect the changes of BMSCs migration after over-expression of PTEN gene.Results The recombinant adenovirus vector was identified by restriction endonuclease digestion.After over-expression of PTEN gene,the migration of BMSCs was inhibited significantly.Conclusion Over-expression of PTEN gene significantly inhibits the migration of BMSCs,which lay a foundation for further study on the migration mechanism of PTEN gene to BMSCs.