成纤维细胞生长因子在软骨发育过程中的基因芯片分析及相关研究  

Expression profile of fibroblast growth factor in MAPK signaling pathway during chondrogenesis in mice

作　　者：唐欣[1] 徐忠世[1] 杨述华[1] 陈雨辰[2] 李奇[2] 余从年[3] 杨操[1] 李进[1] 许伟华[1] 

机构地区：[1]华中科技大学附属协和医院骨科,武汉430022 [2]生物芯片上海国家工程研究中心 [3]华中科技大学同济医学院医学生物学系

出　　处：《中华创伤骨科杂志》2009年第4期362-365,共4页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金（3471753）;湖北省自然科学基金（2008CDB197）

摘　　要：目的榆测并分析促分裂原活化蛋白激酶（MAPK）信号通路中成纤维细胞生长因子（FGF）在软骨发育过程中的基囚表达规律；通过细胞培养观察FGF基因对骨髓基质干细胞（BMSCs）生长特性的影响。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MAPK信号通路中碱性成纤维细胞生长因子（bFGF）在软骨发育过程中的基因表达规律。构建bFGF质粒并转染至培养的BMSCs中，用MTT法、免疫组织化学、HE染色、RT—PCR及酶联免疫吸附法检测bFGF基因转染BMSCs的效果及产物表达。结果MAPK信号通路中的FGF在软骨发育过程中的软骨形成关键期表达显著上调，并肩动MAPK信号通路，促进软骨形成。bFGF基因转染的BMSCs生长活力较强，可以保持2周以上；HE染色显示细胞增殖旺盛，胞核深染；RT—PCR表明有bFGF的基因表达；酶联免疫吸附法检测bFGF表达量高。结论FGF能够启动MAPK信号通路从而促进软骨形成。bFGF质粒转染BMSCs后可促进BMSCs的增殖，细胞有向软骨细胞分化趋势。Objective To explore the gene expression profile offibroblast growth factor （FGF） in MAPK signaling pathway during entochondrostosis in mice and investigate the effects of transfection of pcDNA-bFGF on mice bome marrow stromal cells （BMSCs） in vitro. Methods cDNA microarray technique with 34, 000 genes was used to analyze the gene expression profile during entochondrostosis in the limbs of mice embryo from E10 to E14. Pathway analysis of FGF in MAPK signaling pathway was performed with GCOS1.2 software. The recombined expression vector pcDNA-bFGF was constructed and transfected into mice BMSCs by Lipofectamine. The phenotype changes of cells were observed by cell energometry, HE stain and immunohistochemical assay under light microscopy, RT-PCR and Elisa. Results The gene expression of FGF in MAPK signaling pathway during the critical phase of chondrogenesis in the limbs of mice embryo at E12 was increased obviously. FGF started the MAPK signaling pathway and promoted chondrogenesis. The phenotypes of BMSCs were verified significantly except the bFGF transfected group. As compared with the vector and blank groups, the bFGF transfected group had a tendency of chondrocyte cytodifferentiation. Conclusions FGF may start the MAPK signaling pathway and promote chondrogenesis. The transfection of bFGF gene into mice BMSCs can help BMSCs differentiate into chondrocytes, which should be further investigated for cartilage tissue engineering.

关 键 词：成纤维细胞生长因子 软骨 寡核苷酸序列分析 骨髓细胞 转染 

分 类 号：R686[医药卫生—骨科学]