中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (10): 1578-1583.doi: 10.12307/2023.319

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

TLN1通过激活β-catenin信号通路负性调控骨髓间充质干细胞的成脂分化

黄红颜1,柯皓腾2,叶伟佳2,邓华宗2,王  涛2,蔡明希2,陈奇凡2,岑水忠2   

  1. 南方医科大学珠江医院,1乳腺外科,2脊柱外科,广东省广州市   510280
  • 收稿日期:2022-04-16 接受日期:2022-06-10 出版日期:2023-04-08 发布日期:2022-09-07
  • 通讯作者: 岑水忠,博士,医师,南方医科大学珠江医院脊柱外科,广东省广州市 510280
  • 作者简介:黄红颜,女,1990年生,河南省开封市人,汉族,2019年重庆医科大学毕业,博士,助理研究员,主要从事干细胞的基础研究、乳腺癌骨转移、长非编码RNA等基础及临床转化研究。
  • 基金资助:
    广东省基础与应用基础研究基金自然科学基金面上项目(2022A1515012021),项目负责人:岑水忠;广东省基础与应用基础研究基金青年项目(2020A1515110930),项目负责人:岑水忠;南方医科大学珠江医院院长基金青年培育项目(yzjj2020qn06),项目负责人:岑水忠

TLN1 negatively regulates adipogenic differentiation of bone marrow mesenchymal stem cells through activating the beta-catenin signaling pathway

Huang Hongyan1, Ke Haoteng2, Ye Weijia2, Deng Huazong2, Wang Tao2, Cai Mingxi2, Chen Qifan2, Cen Shuizhong2   

  1. 1Department of Breast Surgery, 2Department of Spinal Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
  • Received:2022-04-16 Accepted:2022-06-10 Online:2023-04-08 Published:2022-09-07
  • Contact: Cen Shuizhong, MD, Physician, Department of Spinal Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
  • About author:Huang Hongyan, MD, Assistant researcher, Department of Breast Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong Province, China
  • Supported by:
    the Natural Science Foundation of Guangdong Basic and Applied Basic Research Foundation (General Program), No. 2022A1515012021 (to CSZ); the Natural Science Foundation of Guangdong Basic and Applied Basic Research Foundation (Youth Program), No. 2020A1515110930 (to CSZ); the Dean's Fund Youth Development Program of Zhujiang Hospitial, Southern Medical University, No. yzjj2020qn06 (to CSZ)

摘要:

文题释义:
TLN1:TLN1编码的Talin1是一种细胞内的细胞骨架蛋白,是黏附复合体的主要成分之一,对整合素的活化至关重要,可促进整合素与肌动蛋白骨架之间的连接,在细胞功能中发挥关键调控作用。
β-catenin信号通路:可调节干细胞的多能性,在发育过程中决定细胞的分化命运,也被认为与多种癌症的发病机制之间关系密切,该通路已成为重要的癌症药物靶点。此外,该通路在骨髓间充质干细胞成骨-成脂平衡中发挥关键作用,其中β-catenin信号通路激活可抑制骨髓间充质干细胞成脂分化,因此探索β-catenin信号通路上游靶点和调控机制具有重要意义。

背景:骨髓间充质干细胞成骨-成脂分化平衡在骨代谢疾病中发挥关键作用,课题组前期研究证实TLN1可调控骨髓间充质干细胞成骨分化,但其对骨髓间充质干细胞成脂分化的作用仍未明确。
目的:探讨TLN1在骨髓间充质干细胞成脂分化中的作用及机制。
方法:诱导骨髓间充质干细胞成脂分化并使用Western blot及qRT-PCR检测TLN1的表达变化;采用CCK-8检测TLN1敲减后骨髓间充质干细胞增殖能力变化,成脂诱导后进行油红O染色及定量分析,qRT-PCR检测骨髓间充质干细胞成脂分化的关键分子PPAR-γ、C/EBP-α的基因表达水平;Western blot检测成脂经典通路蛋白的表达,同时在敲减TLN1的同时添加β-catenin激活剂后检测骨髓间充质干细胞的成脂分化能力,以探讨TLN1调节成脂分化的具体分子机制。
结果与结论:TLN1在骨髓间充质干细胞的成脂分化过程中表达下调;敲减TLN1对骨髓间充质干细胞的增殖无明显影响,可显著增强骨髓间充质干细胞成脂诱导分化能力;敲减TLN1可抑制活性的β-catenin表达,而激活β-catenin信号通路可逆转TLN1敲减介导的成脂分化增强。结果表明,TLN1表达在骨髓间充质干细胞成脂分化过程中逐渐下调,敲减TLN1通过抑制β-catenin信号通路发挥促进骨髓间充质干细胞成脂分化的作用。

https://orcid.org/0000-0003-2339-7189 (黄红颜);https://orcid.org/0000-0001-8499-1320 (岑水忠) 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关键词: 间充质干细胞, TLN1, 成脂分化, β-catenin, 信号通路, 组织工程

Abstract: BACKGROUND: The balance between osteogenic and adipogenic differentiation in bone marrow mesenchymal stem cells plays a key role in bone metabolic diseases. Our previous study confirmed that TLN1 can regulate osteogenic differentiation of bone marrow mesenchymal stem cells, but its effect on adipogenic differentiation of bone marrow mesenchymal stem cells remains unclear. 
OBJECTIVE: To investigate the role and mechanism of TLN1 in the adipogenic differentiation of bone marrow mesenchymal stem cells. 
METHODS:  Bone marrow mesenchymal stem cells were induced to adipogenic differentiation and TLN1 expression was detected by qRT-PCR and western blot assay. CCK-8 was used to detect the proliferation of bone marrow mesenchymal stem cells after TLN1 knockdown. The Oil red O staining and quantification were performed after adipogenic induction. Besides, qRT-PCR was used to detect the gene expression levels of the key molecules of adipogenic differentiation of bone marrow mesenchymal stem cells including PPAR-γ and C/EBP-α. Western blot assay was used to detect the classic pathway of adipogenesis. Meanwhile, TLN1 was knockdown and β-catenin activator was added to detect the adipogenic differentiation of bone marrow mesenchymal stem cells, to explore the specific molecular mechanism of TLN1 regulating adipogenesis. 
RESULTS AND CONCLUSION: TLN1 expression was down-regulated during adipogenic differentiation of bone marrow mesenchymal stem cells. TLN1 knockdown had no significant effect on the proliferation of bone marrow mesenchymal stem cells, but significantly increased the adipogenic induction of bone marrow mesenchymal stem cells. TLN1 knockdown could inhibit the expression of active β-catenin, while activation of β-catenin signaling pathway could reverse the enhancement of adipogenesis mediated by TLN1 knockdown. These findings suggest that TLN1 expression is gradually down-regulated during the adipogenic differentiation of bone marrow mesenchymal stem cells, and TLN1 knockdown can promote the adipogenic differentiation of bone marrow mesenchymal stem cells by inhibiting β-catenin signaling pathway.

Key words: mesenchymal stem cell, TLN1, adipogenic differentiation, β-catenin, signaling pathway, tissue engineering

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