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在缺乏DNA甲基转移酶3B(DNMT3B)的情况下,脐间充质干细胞通过上调Runx2表达而增强成骨作用。

Osteogenesis of umbilical mesenchymal stem cells is enhanced in absence of DNA methyltransferase 3B (DNMT3B) through upregulating Runx2 expression.

作者信息

Zhu X-W, Zuo J-L, Liu Y-H, Zang R, Li Y-K, Wang X, Li J-M

机构信息

Shandong University, Jinan, China.

出版信息

Eur Rev Med Pharmacol Sci. 2014 Oct;18(20):3004-9.

Abstract

OBJECTIVE

Umbilical mesenchymal stem cells (UMSCs) is one of most popular regenerative medical source of bone replacement therapy in both clinical and scientific researches. However, it is still low effective to induce the osteogenesis of hUMSCs. In this study, we aimed to elucidate the roles of DNA methyltransferase 3B (DNMT3B) in the osteogenesis of hUMSCs.

MATERIALS AND METHODS

Knockdown DNMT3B in hUMSCs was gained via RNA interference technology. After confirming the decrease of DNMT3B in mutant hUMSCs by immunostaining and qPCR, osteogenesis differentiation was carried out. To identify the phenotype of osteogenesis in both bone formation ability and function of bone, immunostaining, qPCR and functional test was performed, compared to wildtype hUMSCs.

RESULTS

Real-time Quantitative PCR (qPCR) and immunostaining results indicated that lacking of DNTM3B the osteogenesis related genes were significantly downregulated. Meanwhile, the functional test was also consistent with the downregulated differentiation result.

CONCLUSIONS

The osteogenesis differentiation of hUMSCs is impaired in the absence of DNMT3B.

摘要

目的

脐间充质干细胞(UMSCs)是临床和科研中骨替代治疗最常用的再生医学来源之一。然而,诱导人脐间充质干细胞成骨的效率仍然较低。在本研究中,我们旨在阐明DNA甲基转移酶3B(DNMT3B)在人脐间充质干细胞成骨过程中的作用。

材料与方法

通过RNA干扰技术敲低人脐间充质干细胞中的DNMT3B。通过免疫染色和qPCR确认突变型人脐间充质干细胞中DNMT3B减少后,进行成骨分化。与野生型人脐间充质干细胞相比,通过免疫染色、qPCR和功能测试来鉴定骨形成能力和骨功能方面的成骨表型。

结果

实时定量PCR(qPCR)和免疫染色结果表明,缺乏DNTM3B时,成骨相关基因显著下调。同时,功能测试也与分化下调结果一致。

结论

在缺乏DNMT3B的情况下,人脐间充质干细胞的成骨分化受损。

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