X线照射对MC3T3一E1成骨细胞.PDFVIP

生堡匡堂盘查!Q!垒生!旦!旦筮!鱼鲞筮!!翅塑!!!丛鲤』￡!i塑：坠E堕坐!竺!：!!!鱼：!些!!：塑!：!! O．5 Gy X线照射对MC3 T3一E1成骨细胞 基因表达谱的影响 陈明 董启榕黄群徐炜 余昶 【摘要】 目的探索低剂量x线促进成骨分化的关分子机制。方法0．5 Gy X线照射成骨细 胞后继续诱导成骨培养，采用基因芯片技术检测x线照射成骨细胞后基因表达谱的变化，筛检出表 达差异基因，选取部分骨分化相关基因通过实时聚合酶链反应(real-time PCR)进行验证，并进行基因 本体(Gene Ontology，GO)分析和通路(Pathway)分析。结果成骨细胞在低剂量x射线照射后，共发 现1 412个差异表达基因，其中559个上调，853个下调。Real—time PCR结果显示，0．5 Gy x线照射组 mRNA表达水平显著升高的基因包括：Bglap-rsl(2．5倍)、Colla2(2．1倍)、Td-131(3．4倍)、Lrp5(2．5 倍)、Dvll(4．1倍)、Map4k5(1．6倍)(P0．01)，而Id2(0．5倍)、Dkkl(0．2倍)表达水平显著降低 (P0．01)。免疫印迹结果显示0．5 Gy X线照射成骨细胞后TGF-131、LRP-5蛋白表达增加(P 0．05)，Dkkl蛋白表达减低(P0．01)。GO和Pathway分析结果显示细胞外信号、细胞黏附斑信号、 Wnt信号、IKB／NF．KB信号、生长因子受体信号明显富集。结论低剂量x线照射成骨细胞后，可能 通过细胞外信号、细胞黏附斑信号、Wnt信号、IKB／NF-KB信号、生长因子受体信号通路促进成骨细 胞分化和矿化，这些信号分子可以相互作用形成复杂的网络体系，最终导致成骨细胞表型特征的改 变性。 【关键词】x线；成骨细胞；寡核苷酸类；序列分析 基金项目：国家自然科学基金 Effects of 0．5 Gy X-ray radiation on the proffie of gene expression in MC3T3-E1 osteoblasts Chen 胧昭，Dong Qirong，Huang Qua，舰耽i，She Chang．Department of Orthopedics，the Second Hospital Affiliated to Soochow University，Suzhou 215004，China Corresponding author：Dong Qirong，Email：由r@szgla net 【Abstract】 Objective To investigate the molecular mechanism of low—dose X-ray irradiation on osteoblasts by detecting the gene expression profiles of osteoblasts radiated with 0．5 Gy X·ray．Methods Mieroarray analyses to value the changes of gene expression of MC3T3一E1 osteoblasts after 0．5 Gy X—ray irradiation were conducted．The end points included modulation of key markel苫，and pathway and gene ontology through transcriptomic profiling and bioinformatics analysis．Several major genes during osteoblasts differentiation were selected for Real-time PCR analysis．Results l 412 differentially expressed transcripts were identified between the radiated group and control group．Among the identified genes，559 transcripts were uD—regulated and 853 transcripts were down—regulated after irradiation．Real．time PCR analysis indicated that the mRNA expressions of Bglap—rsl，Coll a2，Tgf-131，Lr#，Dvll，Map4k5 were appar