摘要
目的探究miR-26b-5p通过抑制抑制元素1-沉默转录因子(repressor element 1-silencing transcrip-tion factor,REST)表达调节胶质瘤细胞生长和侵袭情况.方法逆转录-聚合酶链反应检测miR-26b-5p和REST表达水平;荧光素酶报告实验检测miR-26b-5p和REST的靶向关系;Hoechst染色检测细胞凋亡;Colony形成法检测细胞增殖;Transwell检测细胞侵袭;蛋白免疫印迹检测细胞Ki67、MMP-9、cleaved caspase-3蛋白表达水平.结果miR-26b-5p和REST存在直接靶向作用关系,与Control组比较,mimic组细胞增殖数、侵袭数和Ki67、MMP-9表达明显降低,凋亡率与cleaved caspase-3表达明显升高,inhibitor组细胞增殖数、侵袭数和Ki67、MMP-9表达明显升高,凋亡率与cleaved caspase-3表达明显降低.结论miR-26b-5p可以通过抑制对REST的表达调节胶质瘤细胞生长、侵袭、凋亡能力.
Objective To investigate the role of miR-26b-5p in regulating glioma cell growth and invasion by inhibiting REST expression.Methods miR-26b-5p and REST expression levels were detected by reverse transcription-polymerase chain reaction.Luciferase report assay was used to detect the targeting relationship between miR-26b-5p and REST.Hoechst staining was used to detect apoptosis.Colony formation method was employed to measure cell proliferation,Transwell to detect cell invasion and Western blotting to detect the expression levels of Ki67,MMP-9 and cleaved caspase-3.Results miR-26b-5p showed a direct targeting relationship with REST.The number of proliferative cells,the number of invasive cells,and the expression levels of Ki67 and MMP-9 were significantly decreased and the apoptosis rate and cleaved caspase-3 expression were significantly in-creased in mimic group when compared with those in the control group.The number of proliferative cells,the number of invasive cells,and the expression levels of Ki67 and MMP-9 were significantly increased,while the apoptosis rate and cleaved caspase-3 expression were significantly decreased in the inhibitor group as compared with those in the control group.Conclusion miR-26b-5p can regu-late the growth,invasion and apoptosis of glioma cells by inhibiting the expression of REST.
作者
邱锋
王荟
龚立
陈佳
蔡佩浩
QIU Feng;WANG Hui;GONG Li;CHEN Jia;CAI Peihao(Department of Neurosurgery,Shuguang Hospital Affiliated to University of Traditional Chinese Medicine,Shanghai,201203,China;Department of Pathology,Shuguang Hospital Affiliated to University of Traditional Chinese cine,Shanghai,201203,China)
出处
《医学分子生物学杂志》
CAS
2020年第1期40-45,共6页
Journal of Medical Molecular Biology
基金
上海市卫生和计划生育委员会科研课题项目(No.201640393)。