基因芯片筛选骨髓间充质干细胞与透明软骨细胞的差异表达基因

Gene chips select differential representing genes of mesenchymal stem cells and hyaline chondrocytes

关键词:  基因芯片  骨髓间充质干细胞  透明软骨  弹性软骨

Key words:  gene chip  mesenchymal stem cells  hyaline cartilage  elastic cartilage

摘要:

［目的］通过筛选大鼠骨髓间充质干细胞（mesenchymal stem cells，MSCs）与透明软骨细胞及弹性软骨细胞的差异表达基因，为进一步筛选可诱导MSCs向透明软骨细胞分化的生长因子打下基础。［方法］分离获得所需MSCs、弹性软骨细胞和透明软骨组织，提取细胞和组织块的总RNA并纯化。获得经荧光标记的cDNA，与基因芯片杂交后扫描荧光信号图像，对所获图像进行分析，找到差异表达基因。分析所获差异表达基因功能。 ［结果］ 以2倍差异显著性为标准,在两种软骨中共同上调的基因数为582条，而于透明软骨组中上调，弹性软骨组中下调的基因数为459条。将基因功能查询结果与cy5/cy3比值大小结合考虑后，确定了软骨调节素1（chondromodulin1，Chm1）基因、前II型胶原α1链(procollagen，type2，alpha1， Col2α1) 基因和软骨同源异型蛋白1（Cartilagehomeoprotein 1，Cart1）基因可做为目标研究基因。［结论］Chm1基因、Col2α1基因与Cart1基因可能是诱导MSCs向透明软骨细胞方向分化的关键基因。

Abstract:

［Objective］To lay a foundation work for selecting the new growth factors which were able to induce the differentiation from MSCs to hyaline chondrocytes through selecting the differential representing genes of rat mesenchymal stem cells, hyaline chondrocytes and elastic chondrocytes.［Methods］The needed MSCs, elastic chondrocytes and hyaline cartilage tissues were acquired through segregation, and the general RNA was extracted from the cells and tissue lumps. And the cross between the acquired cDNA which was marked by fluorescence and gene chip was done. The fluorescent signal images were scanned and analyzed, then the differential representing genes were found and functionally analyzed.［Results］Using the twice significance of difference as the standard, totally there were 582 same up-regulated genes in botto of groups, while the number of genes which were up-regulated in the hyaline chondrocytes group and down-regulated in the elastic chondrocytes group were 459.Considering the function and the ratio of cy5 to cy3, Chm1, Col2α1 as well as Cart1 were designated as the target genes.［Conclusions］Chm1,Col2α1 and Cart1 might be the key factors in inducing the differentiation from MSCs to hyaline chondrocytes.