摘要
该研究探讨了抑制Notch信号通路联合沉默Id1对人骨肉瘤细胞MG63的恶性生物学行为及成骨分化的影响。采用Notch信号通路抑制剂DAPT、沉默Id1重组腺病毒分别或联合处理MG63细胞,采用Western blot检测分组处理MG63细胞后Notch1、Jagged1、Id1蛋白的表达;CCK8检测分组处理后MG63增殖能力;流式细胞术检测分组处理后MG63细胞凋亡水平;划痕实验和Transwell检测分组处理后MG63细胞迁移和侵袭能力;碱性磷酸酶、茜素红染色分别检测分组处理后MG63细胞早期、晚期成骨分化能力。结果表明,DAPT处理MG63细胞后,Notch1、Jagged1蛋白表达下调(P<0.05),可有效抑制MG63细胞中Notch信号通路活性;抑制MG63细胞中Notch信号通路后Id1蛋白水平表达下降,抑制MG63细胞中Notch信号通路联合沉默Id1后Id1蛋白表达水平最低(P<0.05);抑制MG63细胞中Notch信号通路后细胞增殖、迁移、侵袭能力下降,凋亡水平增加和早期成骨分化能力减弱(P<0.05);抑制MG63细胞中Notch信号通路联合沉默Id1后细胞增殖、迁移、侵袭能力进一步减弱,凋亡水平最高,早期、晚期成骨分化能力增强(P<0.05)。综上所述,抑制Notch信号通路可减弱MG63细胞恶性;抑制Notch信号通路联合沉默Id1后可进一步减弱MG63细胞恶性,促进其成骨分化。
The aim of the research is to study effect of inhibiting Notch signaling pathway combined with silencing Id1 on biological behavior and osteogenic differentiation of human osteosarcoma cell MG63.MG63 cells were treated with the Notch signaling pathway inhibitor DAPT and silencing Id1 alone or jointly.The expression of Notch1,Jagged1,Id1 in each group was detected by Western blot.CCK8 was used to detect the proliferation ability of MG63 cells in different groups.Flow cytometry was used to detect the apoptosis level of MG63 cells in different groups.Scratch test and Transwell were used to detect the migration and invasion abilities of MG63 cells in different groups.Alkaline phosphatase and Alizarin red staining were used to detect the early and late-stage osteogenic differentiation abilities of MG63 cells in different groups,respectively.The results indicate that DAPT can inhibit the Notch signaling pathway with the down-regulated expression of Notch1 and Jagged1 (P<0.05).The Id1 expression decreased after inhibiting the Notch signaling pathway of MG63 cells,and the Id1 expression level was lowest after inhibiting the Notch signaling pathway combined with silencing Id1 of MG63 cells (P<0.05).The proliferation,migration,invasion ability and the early-stage osteogenic differentiation ability decreased,but the apoptosis level increased after inhibiting the Notch signaling pathway of MG63 cells (P<0.05).After inhibiting the Notch signaling pathway combined with silencing Id1 of MG63 cells,the proliferation,migration and invasion ability decreased further,but the osteogenic differentiation ability increased,meanwhile the apoptosis level was the highest (P<0.05).To sum up,inhibiting the Notch signaling pathway can weaken MG63 cells malignancy.While,inhibiting the Notch signaling pathway combined with silencing Id1 can further weaken the malignancy of MG63 cells.It can also promote MG63 osteogenic differentiation.
作者
陈洁
张瑶
谢圣男
周红
罗庆
CHEN Jie;ZHANG Yao;XIE Shengnan;ZHOU Hong;LUO Qing(Department of Pediatric Research Institute of Children’s Hospital of Chongqing Medical University,National Clinical Research Center for Child Health and Disorders(Chongqing),Ministry of Education Key Laboratory of Child Development and Disorders,China International Science and Technology Cooperation Base of Child Development and Critical Disorders,Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2021年第4期715-724,共10页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:81172545)资助的课题。
作者简介
通讯作者:罗庆,Tel:13452336031,E-mail:352934430@qq.com。
