[1]赖桂华,黄爱兰,赵志,等.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,(05):505.
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MircoRNA-218下调BMI-1表达水平介导的抗骨肉瘤作用()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2018年05期
页码:
505
栏目:
出版日期:
2018-05-15

文章信息/Info

Title:
MicroRNA-218 promotes osteosarcoma cell apoptosis by downregulating oncogene B lymphoma mouse Moloney leukemia virus insertion region 1
作者:
赖桂华黄爱兰赵志卢兴浩祖文轩
关键词:
miR-218BMI-1骨肉瘤凋亡
Keywords:
miR-218 oncogene B lymphoma mouse Moloney leukemia virus insertion region 1 osteosaroma apoptosis
摘要:
目的初步探讨microRNA-218在人骨肉瘤中的抗肿瘤作用及其分子机制。方法利用qRT-PCR检测68例人骨肉瘤组织 和癌旁组织中miR-218 的表达水平。将miR-218 模拟物或抗miR-218 模拟物转染入人骨肉瘤Saos-2 细胞,通过CCK-8、 Annexin V-FITC和Western blotting检测转染后的细胞活性、细胞凋亡和C-PARP蛋白表达水平。利用luciferase assay筛选和识 别miR-218的作用靶点及具体作用位点,进一步通过qRT-PCR 和Western blotting检测转染miR-218模拟物或抗miR-218模拟 物后Saos-2细胞中BMI-1基因和蛋白的表达水平。结果人骨肉瘤组织中的miR-218表达水平明显低于癌旁组织。CCK-8结 果显示,与对照组相比,转染miR-218模拟物24、36和48 h后的Saos-2细胞活性显著降低,而转染抗miR-218模拟物的Saos-2细 胞活性则显著增高。同时,转染miR-218模拟物组的C-PARP的蛋白表达水平较转染抗miR-218模拟物组和对照组明显增强。 Annexin V-FITC凋亡检测结果显示,转染miR-218模拟物组的细胞凋亡数和凋亡率较对照组显著增加。此外,luciferase assay 结果显示Saos-2细胞中miR-218的特异性作用靶点为BMI-1,在miR-218过表达时BMI-1的基因和蛋白表达水平明显被抑制, 而miR-218 敲除时则显著增强;miR-218 可通过直接结合BMI-1 3’-UTR抑制BMI-1 的表达。结论miR-218 可能通过下调 BMI-1水平促进人骨肉瘤细胞Saos-2细胞凋亡,进而发挥其抗肿瘤作用。
Abstract:
Objective To investigate the tumor-suppressing effect of microRNA-218 (miR-218) in osteosarcoma (OS) and explore its molecular mechanism. MethodsWe 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 andWestern blotting.We also analyzed the potential functional targets of miR-218 in Saos-2 cells using luciferase assay, qRT-PCR andWestern blotting. Results The expression level of miR-218was 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.

相似文献/References:

[1]李超,涂康生,郑鑫,等.MicroRNA-218在肝细胞癌中的表达及功能[J].南方医科大学学报,2013,(08):1127.
[2]毛楠,何关生,饶进军,等.沉默Bmi-1表达可逆转肺癌细胞对顺铂的耐药性[J].南方医科大学学报,2014,(07):1000.

更新日期/Last Update: 1900-01-01