Role of PTEN in bone morphogenetic protein 9 induced osteogenic differentiation in mouse embryonic fibroblasts

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Role of PTEN in bone morphogenetic protein 9 induced osteogenic differentiation in mouse embryonic fibroblasts

更新时间：2026-01-09

- Role of PTEN in bone morphogenetic protein 9 induced osteogenic differentiation in mouse embryonic fibroblasts
- Journal of Army Medical University Vol. 38, Issue 3, Pages: 264-269(2016)
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  CLC： R322.71;R329-33;R329.2
- Published：2016
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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.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,Cheng Qianzhao,Yuan Shuangxue,Wang Dongxu,Zeng Yuhua,Li Yang,Shao Ying,Huang Jun,He Baicheng.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.
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.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. DOI：

　Wu Qiuxiang,Ren Chunmei,Chen Zhenhua,Cheng Qianzhao,Yuan Shuangxue,Wang Dongxu,Zeng Yuhua,Li Yang,Shao Ying,Huang Jun,He Baicheng.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. DOI：

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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