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[同源盒基因在硬组织发育过程中的分子克隆与表达]

[Molecular cloning and expression of homeobox-containing genes during hard tissue development].

作者信息

Iimura T

机构信息

Department of Biochemistry, Faculty of Dentistry, Tokyo Medical and Dental University.

出版信息

Kokubyo Gakkai Zasshi. 1994 Dec;61(4):590-604. doi: 10.5357/koubyou.61.590.

DOI:10.5357/koubyou.61.590
PMID:7897272
Abstract

Although much is known about the hormonal regulation of hard tissue development, much less is known about the nuclear regulatory molecules that affect the process. Homeobox-containing genes are thought to encode DNA-binding transcriptional factors which control the expression of other genes. In this study, molecular cloning of Msx homeobox-containing genes from a bovine odontoblast library and a human dental pulp-derived cells library was performed, and also the expression of mRNAs for Hox and Msx homeobox-containing genes during ectopic bone formation induced by BMP was investigated. Screening of a bovine odontoblast cDNA library and a human dental pulp-derived cells library with murine Msx-1 and Msx-2 cDNA probes led to the isolation of several positive clones. All of the clones from a bovine odontoblast library encoded the bovine counterpart for human MSX-1. All of the clones from a human dental pulp-derived cells library encoded the human counterpart for murine Msx-2. Northern blot analysis probed with a human MSX-2 cDNA indicated the expression of 2.2kb and 1.2kb mRNA in human dental pulp-derived cells. Polymerase chain reaction (PCR)-based analysis in the BMP-implanted tissue revealed that nine rat homologues of Hox homeobox-containing genes were expressed in the early cell migration stage and that two Msx genes were expressed in the cartilage and bone differentiation stages. The PCR study provided evidence of dynamic changes in the BMP-induced homeobox gene expression.

摘要

尽管人们对硬组织发育的激素调节了解很多,但对影响这一过程的核调节分子却知之甚少。含同源盒的基因被认为编码控制其他基因表达的DNA结合转录因子。在本研究中,从牛成牙本质细胞文库和人牙髓来源细胞文库中对含Msx同源盒的基因进行了分子克隆,并且研究了BMP诱导的异位骨形成过程中Hox和含Msx同源盒基因的mRNA表达。用鼠Msx-1和Msx-2 cDNA探针筛选牛成牙本质细胞cDNA文库和人牙髓来源细胞文库,得到了几个阳性克隆。牛成牙本质细胞文库中的所有克隆都编码了人类MSX-1的牛对应物。人牙髓来源细胞文库中的所有克隆都编码了鼠Msx-2的人类对应物。用人MSX-2 cDNA探针进行的Northern印迹分析表明,2.2kb和1.2kb的mRNA在人牙髓来源细胞中表达。基于聚合酶链反应(PCR)对BMP植入组织的分析显示,九个含Hox同源盒基因的大鼠同源物在早期细胞迁移阶段表达,两个Msx基因在软骨和骨分化阶段表达。PCR研究为BMP诱导的同源盒基因表达的动态变化提供了证据。

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