SOX9基因在软骨发育过程中的表达谱分析及其质粒转染鼠骨髓基质细胞的实验研究

Assessment of the expression profile of SRY-type high-mobility-group box-9 during chondrogenesis of mice and the effects of transfection of recombinant rat sty-type high-mobility-group box-9 on mice mesenchymal stem cells in vitro

目的检测并分析SOX9基因在软骨发育过程中的表达规律；构建其质粒并转染至培养的骨髓源基质细胞（BMSCs）中，通过细胞培养观察SOX9基因对BMSCs生长特性的影响，为可能的骨关节炎（OA）基因治疗提供理论基础。方法用基因芯片技术建立妊娠胎鼠肢芽软骨发育过程的基因表达谱，分析SOX9基因在软骨发育过程中的表达规律；用噻唑蓝（MTT）法、免疫组织化学、苏木素-伊红（HE）染色、反转录-聚合酶链反应（RT—PCR）及酶联免疫吸附试验（ELISA）法检测SOX9基因转染MSCs的效果及产物的表达。结果SOX9基因在软骨发育过程中的软骨形成关键期表达显著上调；pDC316-SOX-9质粒转染骨髓基质细胞后可促进骨髓基质细胞的增殖，细胞有向软骨细胞分化趋势。结论SOX9能够促进软骨形成，SOX9基因质粒转染的MSCs可望在软骨组织工程临床治疗中得到更广泛的运用。

Objective To examine the gene expression profile of SRY-type high-mobility-group box- 9（SOX9） during entochondrostosis of mice and investigate the effects of transfection of the pDC316-SOX-9 on mice mesenchymal stem cells （MSCs） in vitro. Methods cDNA microarray technique with 34 000 genes was used to analyze the gene expression profiles during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of SOX9 was performed with GCOS1.2 software. The recombined expression vector pDC316-SOX-9 was constructed and transfected into mice MSCs by lipofectamine. The pbenotype changes of cells were observed with cell energometry, HE stain, immunohistochemical method, RT-PCR and ELISA. Results The gene expression of SOX9 during the critical phase of chondrogenesis in mice embryo limbs at El2 was increased evidently. SOX9 might promot chondrogenesis. As compared with vector and blank group, the chondrocytes of the SOX9 transfected group had the tendency of enhanced differentiation. Conclusion SOX9 may promote chondrogenesis. The transfection of SOX9 gene into mice MSCs can promot MSCs differentiate into chondrocyte, which may provide some experimental data for cartilage histoengineering.