Notch-1 inhibition reduces proliferation and promotes osteogenic differentiation of bone marrow mesenchymal stem cells
Notch-1 inhibition reduces proliferation and promotes osteogenic differentiation of bone marrow mesenchymal stem cells
复制标题
Notch-1抑制可减少骨髓间充质干细胞的增殖并促进成骨分化
DOI：
10.3892/etm.2019.7765
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发表时间：
2019-09-01
影响因子：
2.7
通讯作者：
Zou, Lijin
中科院分区：
文献类型：
--
作者：
He, Ying;Zou, Lijin
低分化和高增殖率是影响骨髓间充质干细胞(BMSC)成瘤的关键因素。本研究旨在探讨Notch信号通路在骨髓间充质干细胞增殖和成骨分化中的作用。将小鼠骨髓间充质干细胞分为对照组、载体组、Notch1-小干扰(Si)RNA组、γ-分泌酶抑制剂组和Notch1-siRNA+γ-分泌酶抑制剂组。用Notch1 siRNA和/或伽马分泌酶抑制剂处理siRNA-Notch1、γ-分泌酶抑制剂组和Notch1-siRNA+γ-分泌酶抑制剂组。治疗结束后，用细胞计数试剂盒检测细胞增殖情况。逆转录-定量聚合酶链式反应和免疫印迹分析检测肿瘤相关因子，包括转化生长因子-β1、c-Myc和P53。诱导BMSC成骨分化，第14、21天茜素红染色。同时检测碱性磷酸酶(AKP)活性。SiRNA-Notch1和γ-分泌酶抑制剂均能抑制骨髓间充质干细胞的增殖，抑制转化生长因子-β1和c-Myc的表达，增加P53的表达。诱导成骨和茜素红染色后，siRNA-Notch1和γ-分泌酶抑制剂组细胞AKP水平明显高于对照组。研究发现，Notch1抑制抑制BMSCs的增殖，促进其成骨分化。
Low differentiation and high proliferation rates are critical factors affecting bone marrow mesenchymal stem cell (BMSC) tumorigenesis. The present study aimed to investigate the role of the Notch signaling pathway in BMSC proliferation and osteogenic differentiation. Mouse BMSCs were divided into control, vector, Notch1-small interfering (si)RNA, gamma-secretase inhibitor, and Notch1-siRNA + gamma-secretase inhibitor groups. The siRNA-Notch1, gamma-secretase inhibitor, and Notch1-siRNA + gamma-secretase inhibitor groups were treated with Notch1 siRNA and/or gamma-secretase inhibitor. Following treatment, cell proliferation was evaluated using a Cell Counting Kit-8. Tumor-related factors, including transforming growth factor (TGF)-beta 1, c-Myc and p53, were detected by reverse transcription-quantitative polymerase chain reaction and western blot analyses. BMSC osteogenic differentiation was induced and the cells were stained with alizarin red at 14 and 21 days. Alkaline phosphatase (AKP) activity was also evaluated. The siRNA-Notch1 and gamma-secretase inhibitor both reduced BMSC proliferation and the expression of TGF-beta 1 and c-Myc and increased the expression of p53. Following the induction of osteogenesis and staining with alizarin red, the level of AKP was significantly higher in cells in the siRNA-Notch1 and gamma-secretase inhibitor groups compared with that in the control group. It was found that Notch1 inhibition reduced proliferation and promoted the osteogenic differentiation of BMSCs.