摘要
目的探讨miR-153对胶质母细胞瘤细胞株U251凋亡的影响,研究miR-153通诱导U251凋亡的具体机制。方法用qRT-PCR检测正常人脑组织、WHOⅣ级神经胶质母细胞瘤中miR-153的表达情况。用qRT-PCR检测转染miR-153mimics 48h后U251中miR-153的表达量,流式细胞仪检测转染miR-153mimics 48h后U251细胞凋亡情况。荧光素酶报告基因系统验证NF-E2相关因子2(Nrf2)是否是miR-153的靶基因,免疫印迹检测转染miR-153mimics 48h后U251中Nrf2的表达水平。结果与正常脑组织组相比,miR-153在胶质母细胞瘤组表达显著降低(P<0.05)。U251细胞转染miR-153mimics 48h后,miR-153表达水平较对照组明显增高(P<0.05),显著促进了U251的凋亡(P<0.05)。荧光素酶报告基因实验表明,miR-153能特异性靶向Nrf2基因。Western blotting试验证实miR-153过表达抑制Nrf2蛋白的表达。结论 miR-153在神经胶质母瘤细胞中通过靶向Nrf2基因起到抑癌基因的作用,为未来在治疗神经胶质母细胞瘤方面提供了新的靶向治疗观念。
Objective To explore the role of miR-153 involved in the apoptosis of glioblastoma U251 cells and study the molecular mechanisms of miR-153 inducing apoptosis. Methods We detected the expression level of miR-153 in normal brain tissues and WHO IV grade glioblastoma tissues by qRT-PCR. The expression of miR-153 was quantified by Real-time PCR 48 h after transfection with miR-153 mimicsin U251. Cell apoptosis was performed by flow cytometry（FCM） in U251 cells 48 h after transfention of miR-153 mimics. Using a dual-luciferase report assay, we need to identify whether NF-E2 related factor 2 （Nrf2） is the direct target of miRNA-153. The expression of Nrf2 was detected by Western blotting experimen ts after trans- fection of miR-153 mimics. Results The miR-153 was significantly down-regulated inglioblastoma tissues when compared to normal brain tissues（P^0.05）. The expression of miR-153 was significantly up-regulatedafter transfection with miR-153 mim- ics（P^0.05） and it significantly induced apoptosis in glioblastoma U251 cells（P^0.05）. We further identified Nrf2 as a direct target of miRNA-153 by a dual-luciferase report assay. Western blotting experiments confirmed that over-expression of miR- 153 could inhibit the endogenous Nrf2 protein level. Conclusion Taken together, our study demonstrated that miR-153 is a tumor suppressor by targeting Nrf2 and may serve as a potential therapeutic target in glioblastoma in the future.
出处
《中国实用神经疾病杂志》
2013年第19期1-3,共3页
Chinese Journal of Practical Nervous Diseases
基金
国家自然科学基金(81070974
8127377)
