人骨髓基质细胞向成骨细胞分化过程中差异表达基因的筛选

为了探讨人骨髓基质细胞（bone marrow stromal cells,BMSCs）向成骨细胞分化过程中差异表达的基因,本实验采用体外培养人BMSCs,诱导向成骨细胞分化。分别选取培养12和21d的细胞作为驱动方（driver）和实验方（tester）,进行抑制消减杂交,构建cDNA消减文库,将挑选出的阳性克隆与GenBank人基因库中己公布的核酸序列进行同源性比较分析。结果表明,从培养21d的BMSCs中,筛查出5个差异基因,与人基因库中己知基因的同源性分别达到90％以上。有兴趣的是,核心蛋白聚糖和Bax inhibitorl在培养2ld的BMSCs中差异表达。RT-PCR检测显示,核心蛋白聚糖基因在培养21d的细胞中高表达,而在12d的细胞中未检测到表达;Bax inhibitorl基因在培养21d细胞中的表达明显高于12d的细胞。

Screening differentially expressed genes in human bone marrow stromal cells at defined stage of differentiation.

To screen differentially expressed genes involved in osteogenic differentiation of human bone marrow stromal cells(BMSCs) at defined stages,subtractive cDNA library was established by means of suppression subtractive hybridization.The BMSCs cultured for 12 and 21 d were used as driver and tester,respectively.A subtract library was successfully constructed and five positive clones were selected from the library.Sequencing analysis and homology comparison showed that the five clones differentially expressed in BMSCs cultured for 21 d were at least 90% homologous with the known genes in human GenBank.It was interestingly found that the osteogenic BMSCs cultured for 21 d differentially expressed decorin and Bax inhibitor 1.RT-PCR was performed to confirm the differentially expressed genes.The results showed that the expression of Bax inhibitor 1 was significantly higher in the cells of 21-day than that of 12-day,while the expression of decorin was only detected in the cells of 21-day.