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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

刘美玲, 石心泉, 周万灏, 刘洪文, 李东, 贾孟春

国家人口计划生育委员会科学技术研究所.北京 100081

摘要

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，该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21 d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人基因库中已公布的核酸序列进行同源性比较分析。结果表明，从培养21 d的BMSCs中，筛查出5个差异基因，与人基因库中已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 inhibitor 1在培养21 d的BMSCs中差异表达。RT--PCR检测显示，核心蛋白聚糖基因在培养21 d的细胞中高表达，而在12 d的细胞中未检测到表达；Bax inhibitor 1基因在培养21 d细胞中的表达明显高于12 d的细胞。

关键词： 骨髓基质细胞; 成骨细胞; 核心蛋白聚糖; 基因

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

Liu Meiling, Shi Xinquan, Zhou Wanhao, Liu Hongwen, Li Dong, Jia Mengchun

National Population Research Institute for Family Planning.Beijing 100081

Abstract

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.

Key words: bone marrow stromal cells;osteoblast;decorin;Gene

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引用本文：

刘美玲, 石心泉, 周万灏, 刘洪文, 李东, 贾孟春. 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[J]. 生理学报 2006; 58 (4): .

Liu Meiling, Shi Xinquan, Zhou Wanhao, Liu Hongwen, Li Dong, Jia Mengchun. Screening differentially expressed genes in human bone marrow stromal cells at defined stage of differentiation. Acta Physiol Sin 2006; 58 (4): (in Chinese with English abstract).