达沙替尼对人骨髓间充质干细胞生物学特性的影响

发布时间：2018-06-12 07:09

本文选题：人骨髓间充质干细胞 + 达沙替尼　；参考：《中国病理生理杂志》2017年06期

【摘要】：目的:在体外探索达沙替尼对人骨髓来源间充质干细胞(hBMSCs)的活力、迁移、细胞周期和凋亡的影响以及潜在的信号通路,以评估达沙替尼在临床应用中对骨髓造血微环境的影响。方法:CCK-8法检测细胞活力;划痕实验检测细胞迁移;流式细胞术检测细胞周期和凋亡;同时采用吖啶橙/溴化乙啶法检测细胞凋亡;酶联免疫吸附实验检测细胞转化生长因子β1(TGF-β1)和肿瘤坏死因子α(TNF-α)的分泌情况;Western blot检测蛋白激酶B(Akt)蛋白的表达和磷酸化以及cleaved caspase-3的蛋白水平。结果:与对照组相比,达沙替尼(1~10nmol/L)抑制hBMSCs的活力和迁移;在随后的实验中使用的浓度为7 nmol/L。达沙替尼促进细胞凋亡,并使更多细胞的周期阻滞在G_1期。此外,hBMSCs TGF-β1和TNF-α的分泌量显著增加。7 nmol/L达沙替尼组cleaved caspase-3的蛋白水平增加,胞内Akt的蛋白量下调且其磷酸化受到抑制。结论:达沙替尼以浓度依赖性的方式抑制hBMSCs的活力和迁移,并促进TGF-β1和TNF-α的分泌,诱导细胞G_1期阻滞和凋亡;达沙替尼可能通过影响胞内Akt蛋白的表达和磷酸化调控上述细胞学行为。
[Abstract]:Aim: to investigate the effects of dasatinib on the activity, migration, cell cycle and apoptosis of human bone marrow derived mesenchymal stem cells (hBMSCs) in vitro, and to evaluate the effect of dasatinib on the hematopoietic microenvironment of bone marrow in clinical application. Methods Cell viability was detected by: 1) CCK-8 assay; cell migration was detected by scratch assay; cell cycle and apoptosis were detected by flow cytometry; apoptosis was detected by acridine orange / ethidium bromide method. Enzyme linked immunosorbent assay (Elisa) was used to detect the secretion of transforming growth factor 尾 1 (TGF- 尾 1) and tumor necrosis factor 伪 (TNF- 伪). Western blot was used to detect the protein expression, phosphorylation and cleaved caspase-3 protein level. Results: compared with the control group, dasatinide (10 nmol / L) inhibited the activity and migration of hBMSCs, and in subsequent experiments the concentration was 7 nmol / L. Dasatinib promoted cell apoptosis and blocked more cell cycle in G _ 1 phase. In addition, the secretion of TGF- 尾 1 and TNF- 伪 of hBMSCs significantly increased the protein level of cleaved caspase-3 in the group of 7 nmol / L dasatinib, the protein content of intracellular Akt was down-regulated and the phosphorylation of TNF- 伪 was inhibited. Conclusion: dassatinib inhibits the activity and migration of hBMSCs in a concentration-dependent manner, promotes the secretion of TGF- 尾 1 and TNF- 伪, and induces G1-phase arrest and apoptosis of hBMSCs. Dassatinib may regulate the above cytological behavior by affecting the expression and phosphorylation of intracellular Akt protein.
【作者单位】：广州市番禺区何贤纪念医院药剂科;暨南大学血液研究所;暨南大学再生医学教育部重点实验室;暨南大学生命科学技术学院生物工程系;广州军区广州总医院血液科;南方医科大学;
【基金】：国家自然科学基金资助项目(No.81270568)
【分类号】：R96

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