南方医科大学学报2018，Vol.38Issue(5)：505-510,6.
MircoRNA-218下调BMI-1表达水平介导的抗骨肉瘤作用
MicroRNA-218 promotes osteosarcoma cell apoptosis by down-regulating oncogene B lymphoma mouse Moloney leukemia virus insertion region 1
摘要
Abstract
Objective To investigate the tumor-suppressing effect of microRNA-218(miR-218)in osteosarcoma(OS)and explore its molecular mechanism.Methods We examined the expression levels of miR-218 in 68 pairs of OS and adjacent tissue samples using qRT-PCR.Cultured human OS cell line Saos-2 was transfected with miR-218 mimics or anti-miR-218 mimics,and the cell apoptosis was assessed using CCK-8 assay,annexin V-FITC staining and Western blotting.We also analyzed the potential functional targets of miR-218 in Saos-2 cells using luciferase assay,qRT-PCR and Western blotting.Results The expression level of miR-218 was lowered by at least 8 folds in OS tissues as compared with the adjacent tissues.In cultured Saos-2 cells,transfection with miR-218 mimics for 24,36,and 48 h resulted in a significant reduction in the cell viability,while transfection with anti-miR-218 mimics significantly increased the cell viability.The cells transfected with miR-218 mimics showed an obviously enhanced expression of cleaved poly (ADP-ribose)polymerase(C-PARP)as compared with the cells transfected with anti-miR-218 mimics and the control cells.Flow cytometry demonstrated obviously increased apoptosis of the cells following miR-218 mimics transfection. We identified the oncogene B lymphoma mouse Moloney leukemia virus insertion region 1(BMI-1)as a specific target of miR-218 in Saos-2 cells. BMI-1 expressions at both the mRNA and protein levels were significantly reduced in Saos-2 cells overexpressing miR-218 but increased in the cells with miR-218 knockdown as compared to the control cells.Luciferase reporter assay indicated that miR-218 directly inhibited the expression of BMI-1 via binding to its 3'-UTR in OS cells.Conclusion miR-218 can promote OS cell apoptosis and plays the role as a tumor suppressor by down-regulating BMI-1.关键词
miR-218/BMI-1/骨肉瘤/凋亡Key words
miR-218/oncogene B lymphoma mouse Moloney leukemia virus insertion region 1/osteosaroma/apoptosis引用本文复制引用
赖桂华,黄爱兰,赵志,卢兴浩,祖文轩..MircoRNA-218下调BMI-1表达水平介导的抗骨肉瘤作用[J].南方医科大学学报,2018,38(5):505-510,6.基金项目
安徽省高校自然科学研究项目重点项目(KJ2017A211) （KJ2017A211）
安徽省高校自然科学研究项目一般项目(KJ2013Z216)Supported by Key Science Research Project Funding of Education Department of Anhui Province(KJ2017A211)and General Science Research Project Funding of Education Department of Anhui Province(KJ2013Z216). （KJ2013Z216）
