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Molecular cytochemical analysis of correlation,between nuclear distribution of transcription factors and transcriptional activation or cell differentiation inductivity

转录因子核分布与转录激活或细胞分化诱导性相关性的分子细胞化学分析

基本信息

批准号：
18590189
负责人：
IZUMI Shin-ichi
金额：
$ 2.41万
依托单位：
Nagasaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

Runx2, a transcription factor plays an important role in cell differentiation of pluripotential undifferentiated mesenchyme into osteoblasts and chondrocytes, and regulates transcription of the target genes by binding to the promoter. Molecular cytochemistry was performed to clarify whether nuclear distribution of Runx2 correlates with the transcription activation and/or DNA binding activity of the transcription factors for the differentiation.1. When bone morphology protein-2 was introduced to pre-osteoblasts to promote the differentiation by increasing transcription activation of Runx2 gene, Runx2 was strongly stained in the euchromatin area in nuclei.2. In fetal cartilage of transgenic mice with CoIII-Akt-Tg-GFP, Runx2 appeared in the nuclei of undifferentiated perichondrial cells and proliferating chondrocytes. The staining was increased in the nuclear matrix of mature chondrocytes, and decreased in the differentiated hypertrophy chondrocytes. Nuclear distribution of Runx2 seemed to correlate with DNA binding changes of Runx2 by Akt and chondrocyte differentiation.3. In tibia of new-born mice of Runx2-Tg, Runx2 and β-catenin appeared in the nuclei of undifferentiated perichondrial cells, and Runx2 was distributed in the nuclear matrix of osteoblasts but not in the osteocytes. In fetal mice of Runx2-/-, neither β-catenin nor osterix was stained in the nuclei of undifferentiated perichondrial cells. Nuclear distribution of Runx2 and β-catenin seemed to be related with the differentiation.4. When Runx2/GFP were induced in C3H10T1/2 pluripotent undifferentiated fibroblasts, the cytoplasmic processes of the cells were decreased and became short. Immunostaining of actin was decreased and membranous β-catenin aggregated in the cytoplasm. Nuclear matrix distribution of Runx2 seemed to be related with induction of the cell differentiation.

转录因子Runx2在多潜能未分化间充质细胞向成骨细胞和软骨细胞分化过程中起重要作用，并通过与启动子结合来调节靶基因的转录。用分子细胞化学方法阐明Runx2的核分布是否与转录因子的转录激活和/或DNA结合活性有关。结论：1.将骨形态蛋白-2引入成骨前细胞，通过提高Runx2基因的转录活性来促进其分化，Runx2在细胞核常染色质区域呈强阳性表达。在CoIII-Akt-TG-GFP转基因小鼠的胎儿软骨中，Runx2出现在未分化的软骨膜细胞和增殖的软骨细胞的胞核中。在成熟软骨细胞的核基质中染色增强，在分化的肥大软骨细胞中染色减弱。Runx2的核分布可能与Akt对Runx2的DNA结合改变和软骨细胞分化有关。在新生鼠胫骨中，Runx2-Tg和β-catenin出现在未分化的软骨膜细胞的细胞核中，Runx2分布在成骨细胞的核基质中，而不分布在骨细胞中。在Runx2-/-胎鼠中，未分化的软骨膜细胞的细胞核内未见β-连环蛋白和Osterix的染色。Runx2和β-catenin的核分布似乎与分化程度有关。在C3H10T1/2多能未分化成纤维细胞中诱导Runx2/GFP后，细胞胞浆突起减少，变短。肌动蛋白免疫染色减弱，膜型β-连环蛋白聚集在胞浆内。Runx2的核基质分布可能与诱导细胞分化有关。