摘要
目的探讨三七总皂苷(PNS)对U2OS人骨肉瘤细胞的增殖、凋亡、侵袭迁移生物学行为的影响及机制。方法培养U2OS人骨肉瘤细胞,采用(0、200、400、600、800、1000)μg/mL PNS处理细胞后,CCK-8法检测骨肉瘤细胞的增殖,异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测细胞凋亡,集落形成实验检测细胞的集落形成能力,TranswellTM侵袭实验检测细胞侵袭、TranswellTM迁移实验和划痕愈合实验检测细胞迁移;Western blot法检测U2OS细胞缺刻基因1(Notch1)、发状分裂增强子1(Hes1)的蛋白表达。结果与对照组相比,PNS可以呈剂量依赖性抑制U2OS细胞的增殖与集落形成,细胞凋亡率增加,细胞侵袭、迁移能力减弱,细胞Notch1及Hes1的蛋白水平降低。结论PNS通过阻断Notch1的信号通路抑制U2OS骨肉瘤细胞的增殖、侵袭、迁移并促进其凋亡。
Objective To explore the effects of Panax notoginseng saponins(PNS) on the proliferation, apoptosis, migration and invasion of osteosarcoma cell line U2OS and its possible molecular mechanism. Methods Human osteosarcoma cell line U2OS was cultured and treated with(0, 200, 400, 600, 800, 1000) μg/mL PNS. The proliferation of U2OS was detected by CCK-8 method. Annexin Ⅴ-FITC/PI double labeling combined with flow cytometry was used to analyze cell apoptosis. Clone formation assay was used to detect the clone formation ability of the cells. TranswellTM invasion assay was performed to detect cell invasion. TranswellTM migration assay and wound-healing assay were used to determine cell migration. Western blotting was used to detect the expression of Notch1 and downstream protein Hes1 in U2OS cells. Results Compared with the control group, PNS could inhibit the proliferation of U2OS cells and the formation of clonal plaques, increase cell apoptosis rate, weaken the ability of migration and invasion and decrease the expression levels of Notch1 and Hes1 in the cells in a dose-dependent manner. Conclusion PNS can significantly inhibit the proliferation, migration and invasion and promote cell apoptosis of U2OS cells by blocking Notch1 signaling pathway.
作者
徐慧
许涛
方梦影
周智敏
马童童
常林
XU Hui;XU Tao;FANG Mengying;ZHOU Zhimin;MA Tongtong;CHANG Lin(Department of Transfusion,School of Laboratory Medicine,Bengbu Medical College,Bengbu Medical College,Bengbu 233030;Clinical Laboratory Diagnosis Center,Bengbu Medical College,Bengbu Medical College,Bengbu 233030;School of Laboratory Medicine,Bengbu Medical College,Bengbu 233030;Department of Clinical Laboratory,Children’s Hospital of Nanjing Medical University,Nanjing 210008,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2021年第8期716-721,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
安徽省教育厅重点项目(KJ2019A0354)
安徽省大学生创新训练项目(201810367025)。
