[1]伍秋香,任春美,陈振华,等.PTEN在骨形态发生蛋白9诱导小鼠胚胎成纤细胞骨向分化中的作用[J].陆军军医大学学报(原第三军医大学学报),2016,38(03):264-269.
 Wu Qiuxiang,Ren Chunmei,Chen Zhenhua,et al.Role of PTEN in bone morphogenetic protein 9 induced osteogenic differentiation in mouse embryonic fibroblasts[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(03):264-269.
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PTEN在骨形态发生蛋白9诱导小鼠胚胎成纤细胞骨向分化中的作用(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第03期
页码:
264-269
栏目:
基础医学
出版日期:
2016-02-15

文章信息/Info

Title:
Role of PTEN in bone morphogenetic protein 9 induced osteogenic differentiation in mouse embryonic fibroblasts
作者:
伍秋香任春美陈振华陈前昭袁霜雪王东旭曾于桦李洋邵英黄军何百成
重庆医科大学药理学教研室,重庆市生物化学与分子药理学重点实验室
Author(s):
Wu Qiuxiang Ren Chunmei Chen Zhenhua Cheng Qianzhao Yuan Shuangxue Wang Dongxu Zeng Yuhua Li Yang Shao Ying Huang Jun He Baicheng

Department of Pharmacology, Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing Medical University, Chongqing, 400016, China

关键词:
骨形态发生蛋白9PTEN小鼠胚胎成纤维细胞成骨分化BMPs/Smads
Keywords:
bone morphogenetic protein 9 phosphatase and tensin homolog deleted on chromosome ten mouse embryonic fibroblasts osteogenic differentiation BMPs/Smads
分类号:
R322.71; R329-33; R329.2
文献标志码:
A
摘要:

目的      研究PTEN对骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)诱导小鼠胚胎成纤维细胞(mouse embryonic fibroblasts, MEFs)成骨分化的影响,并分析PTEN调节BMP9功能的可能机制。      方法      采用定量PCR检测PTEN在间充质干细胞(mesenchymal stem cells,MSCs)中的表达,分析在MEFs中BMP9对PTEN表达的影响。采用组织化学染色法检测碱性磷酸酶(alkaline phosphatase, ALP)活性,PCR检测OPN和OCN的表达水平,茜素红染色检测钙盐沉积,分析PTEN对BMP9诱导MEFs成骨分化的影响;利用荧光素酶报告质粒和蛋白印迹等方法分析PTEN对骨形态发生蛋白(bone morphogenetic proteins,BMPs)/Smads信号转导的影响。      结果      PTEN在几种常见MSCs中均有表达;BMP9在MEFs中明显抑制PTEN的表达。抑制PTEN能促进BMP9诱导MEFs的ALP活性增加,但过表达PTEN对BMP9诱导MEFs的ALP活性有明显抑制作用。抑制PTEN明显促进BMP9在MEFs细胞中诱导的OCN表达,并增强钙盐沉积。报告质粒分析结果显示,抑制PTEN能明显增强BMPR-Smad报告质粒的转录活性,并增加Smad1/5/8的磷酸化水平。      结论      下调PTEN表达可能是BMP9诱导MEFs成骨分化重要途径之一,其机制可能与促进BMPs/Smads信号转导有关。

Abstract:

Objective       To determine the  effect of phosphatase and tensin homolog deleted on chromosome ten (PTEN) on bone morphogenetic protein 9 (BMP9) induced osteogenic differentiation in mouse embryonic fibroblasts (MEFs) and investigate the possible molecular mechanism.       Methods        Real-time PCR was employed to detect the endogenous expression of PTEN in mesenchymal stem cells (MSCs), and the effect of BMP9 on the expression of PTEN in the MEFs. Histochemical staining was used to assay the activity of alkaline phosphatase (ALP), PCR to detect the expression of osteopontin (OPN) and osteocalcin (OCN) induced by BMP9 in MEFs, and Alizarin Red S staining to detect the effect of PTEN on BMP9 induced matrix mineralization. Finally, BMPR-Smad luciferase reporter and Western blot assay were employed to determine the effect of PTEN on BMPs/Smads signaling pathway.       Results      PTEN was expressed in MSCs, and BMP9 inhibited the expression of PTEN in MEFs. Exogenous expression of PTEN inhibited the enhanced ALP activity induced by BMP9, whereas over-expression of PTEN suppressed the induced activity enhancement. Inhibition of PTEN improved the BMP9-induced expression of OCN in MEFs and promoted matrix mineralization. PTEN inhibitor potentiated BMP9-induced activation of BMPs/Smads signaling and phosphorylation of Smad1/5/8 in MEFs.       Conclusion        Down-regulation of PTEN plays an important role in the promotion of osteogenic differentiation induced by BMP9 in MSCs, which may be mediated by enhancing BMPs/Smads signaling transduction initialized by BMP9.

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更新日期/Last Update: 2016-01-27