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| THSD4基因在小鼠间充质干细胞及MC3T3-E1细胞成骨分化中的作用 |
| To investigate the role of THSD4 gene on osteoblast differentiation of mouse mesenchymal stem cells and MC3T3-E1 cells |
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| DOI：10.3969/j.issn.1006-7108.2022.07.010 |
| 中文关键词: THSD4基因 骨质疏松症 成骨分化；间充质干细胞；小鼠 |
| 英文关键词:THSD4 osteoporosis osteoblast differentiation mesenchymal stem cell mice |
| 基金项目:国家自然科学基金（81772374） |
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| 中文摘要: |
| 目的 探讨THSD4基因对小鼠间充质干细胞和MC3T3-E1细胞成骨分化的影响。方法 提取绝经后骨质疏松症患者的骨髓间充质干细胞进行基因测序分析，与骨关节炎患者的骨髓间充质干细胞进行比较，分析基因表达差异。通过提取不同分化阶段的小鼠骨髓间充质干细胞（M-BMSC）及MC3T3-E1细胞的mRNA来检测THSD4 基因以及成骨分化的标志性基因（ALP、Runx2、Osx）的表达水平。通过构建慢病毒表达载体来实现对M-BMSC及MC3T3-E1细胞中THSD4的敲减及过表达，并观察其对M-BMSC及MC3T3-E1细胞成骨分化能力的影响。结果 THSD4基因在绝经后骨质疏松症患者骨髓间充质干细胞中明显下调，且通过KEGG以及GO富集分析发现THSD4基因可能与PI3K-AKT信号通路及Wnt信号通路相关。随着成骨诱导分化时间的延长，THSD4 mRNA和成骨分化标志性基因（ALP、Runx2、Osx）mRNA在MC3T3-E1以及M-BMSC中表达量均逐渐增加。过表达THSD4可以增强MC3T3-E1细胞和M-BMSC的成骨分化能力，而敲减THSD4则减弱了MC3T3-E1细胞和M-BMSC的成骨分化能力。结论 THSD4基因在绝经后骨质疏松症患者骨髓间充质干细胞中明显下调，且THSD4基因可以增强MC3T3-E1细胞以及M-BMSC的成骨分化能力。 |
| 英文摘要: |
| Objective To pinvestigate the effect of THSD4 gene on the osteoblast differentiation of mouse bone mesenchymal stem cells (M-BMSC) and MC3T3-E1 cells. Methods Bone marrow mesenchymal stem cells from postmenopausal patients with osteoporosis were extracted for gene sequencing analysis, and alternations in gene expression were analyzed. The expression levels of THSD4 and osteoblast differentiation marker genes (ALP, Runx2, Osx) were measured by mRNA quantification from M-BMSC and MC3T3-E1 cells extracted at different stages of differentiation. The lentiviral vector was constructed to achieve knockdown or overexpression of THSD4 in M-BMSC and MC3T3-E1 cells in order to observe its effect on osteoblast differentiation. Results THSD4 gene was significantly down-regulated in bone marrow mesenchymal stem cells of postmenopausal osteoporosis patients, and THSD4 may be related to PI3K-Akt and Wnt signaling pathway. The expression levels of THSD4 mRNA and the markers of osteoblast differentiation (ALP, Runx2, OSX) mRNA in MC3T3-E1 and M-BMSC cells increased with the prolonging of osteoblast differentiation time. THSD4 can enhance the ability of osteoblast differentiation in M-BMSC and MC3T3-E1 cells, knockdown of THSD4 weakened the osteogenic differentiation ability of M-BMSC and MC3T3-E1 cells. Conclusions THSD4 gene was significantly down-regulated in bone marrow mesenchymal stem cells of postmenopausal patients with osteoporosis, and THSD4 could enhance the ability of osteoblast differentiation in M-BMSC and MC3T3-E1 cells. |
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