Gene expression profile during osteoblaslic differentiation of human mesenchymal cells

人间充质细胞成骨细胞分化过程中的基因表达谱

基本信息

批准号：
13557124
负责人：
INOUE Ituro
金额：
$ 7.94万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557124/
关键词：
hMSC osteoblast differentiation OPLL siRNA Wntパスウェイ

项目摘要

Ossification of the posterior longitudinal ligament of the spine (OPLL) is the leading cause of myelopathy in Japan, and is diagnosed by ectopic bone formation in the paravertebral ligament OPLL is a systemic high bone mass disease with a strong genetic background To detect genes relevant to the pathogenesis of OPLL, we performed cDNA microarray analysis of systematic gene expression profiles during osteoblastic differentiation of ligament cells from OPLL patienis (OPLL cells), ossification of yellow ligament patients, and non-OPLL controls, and human mesenchymal stem cells (hMSCs) after stimulating with osteogenic differentiation medium (OS). Twenty-nine genes were up-regulated during osteoblastic differentiation in OPLL cells. Zinc finger protein 145 (PLZF) was one of the highly expressed genes during osteoblastic differentiation in all the cells examined. We investigated the roles of PLZF in the regulation of osteoblastic differentiation ofhMSCs and C2C12 cells. siRNA mediated gene-silencing of PLZF resulted in the reduction of osteoblast-specific genes such as alkaline phosphatase (ALP), collagen lAl (COL1A1), Runx2/cbfa1 (CBFA 1), and osteocalcin (OCN), even in the presence of OS in hMSCs. The expression of PLZF was unaffected by addition of bone morphogenetic protein 2 (BMP-2) and the expression of BMP-2 was not affected by PLZF in hMSCs. In C2C12 cells, overexpression of PLZF increased the expression of Cbfal and Collal, on the other hand, overexpression of CBFA1 did not affect the expression of Plzf. These findings indicate that PLZE plays important roles in early osteoblastic differentiation as an upstream regulator of CBFA1 and thereby might pailicipate in promoting ossification of spinal ligament cells in OPLL patients.

脊柱后纵韧带骨化（OPLL）是日本脊髓病的主要原因，通过椎旁韧带异位骨形成来诊断。OPLL 是一种具有很强遗传背景的系统性高骨量疾病。为了了解 OPLL 的发病机制，我们对 OPLL patienis 韧带细胞（OPLL 细胞）成骨细胞分化过程中系统基因表达谱进行了 cDNA 微阵列分析，黄韧带骨化患者、非 OPLL 对照以及用成骨分化培养基 (OS) 刺激后的人间充质干细胞 (hMSC)。在 OPLL 细胞的成骨细胞分化过程中，有 29 个基因上调。锌指蛋白 145 (PLZF) 是所有检查细胞中成骨细胞分化过程中高度表达的基因之一。我们研究了 PLZF 在调节 hMSC 和 C2C12 细胞成骨细胞分化中的作用。 siRNA 介导的 PLZF 基因沉默导致成骨细胞特异性基因减少，例如碱性磷酸酶 (ALP)、胶原蛋白 lAl (COL1A1)、Runx2/cbfa1 (CBFA 1) 和骨钙素 (OCN)，即使存在 OS在 hMSC 中。 PLZF 的表达不受添加骨形态发生蛋白 2 (BMP-2) 的影响，并且在 hMSC 中 BMP-2 的表达不受 PLZF 的影响。在C2C12细胞中，PLZF的过表达增加了Cbfal和Collal的表达，而CBFA1的过表达不影响Plzf的表达。这些发现表明 PLZE 作为 CBFA1 的上游调节剂在早期成骨细胞分化中发挥重要作用，因此可能参与促进 OPLL 患者脊柱韧带细胞的骨化。