微RNA-107对尤文肉瘤细胞增殖和体外拟态血管形成的抑制作用及其机制探讨  

Inhibitory effects of microRNA-107 on proliferation and in vitro vasculogenic mimicry of Ewing sarcoma cells and the mechanism

作　　者：陈佳骏[1] 周新[1] 肖前仁 王滕羽 李云云 张中卒[1] 邵高海[1] CHEN Jiajun1;ZHOU Xin1;XlAO Qianren2;WANG Tengyu1;LI Yunyun3;ZHANG Zhongzu1;SHAO Gaohai1

机构地区：[1]重庆医科大学附属永川医院骨科,重庆402160 [2]南昌大学第一附属医院骨科,江西南昌330006 [3]重庆医科大学附属永川医院妇产科,重庆402160

出　　处：《肿瘤》2017年第12期1282-1288,共7页Tumor

基　　金：国家自然科学基金资助项目(编号:81502329)~~

摘　　要：目的:探讨微RNA-107(microRNA-107,miR-107)表达对尤文肉瘤细胞增殖和体外拟态血管形成能力的影响,并分析其可能的分子作用机制。方法:采用实时荧光定量PCR法检测不同尤文肉瘤细胞中miR-107的表达水平。将人工合成的miR-107模拟物片段(miR-107 mimic)转染至尤文肉瘤RD-ES和SK-ES-1细胞中,同时设置转染无关片段的对照组。实时荧光定量PCR法检测转染后尤文肉瘤细胞中miR-107表达的恢复情况。然后,分别采用CCK-8法和基质胶血管生成拟态实验检测miR-107高表达对尤文肉瘤细胞增殖及血管拟态形成能力的影响,并采用实时荧光定量PCR法和蛋白质印迹法检测细胞中低氧诱导因子1β(hypoxia inducible factor-1β,HIF-1β)mRNA及蛋白表达水平的变化。结果:与人间充质干细胞相比,miR-107在尤文肉瘤RD-ES和SK-ES-1细胞中呈低表达(P值均<0.01)。转染miR-107 mimic后,尤文肉瘤RDES及SK-ES-1细胞中miR-107的表达水平明显升高(P值均<0.01)。miR-107过表达后,RD-ES及SK-ES-1细胞的增殖能力明显降低(P值均<0.05),体外血管拟态形成也被明显抑制(P值均<0.05),而且细胞中HIF-1βmRNA及蛋白表达均明显下调(P值均<0.01)。结论:miR-107在尤文肉瘤细胞中低表达。miR-107过表达能够抑制尤文肉瘤细胞的增殖及体外拟态血管形成,该作用可能是通过靶向调控HIF-1β表达来实现的。Objective： To investigate the effects of microRNA-107 （miR-107） on the proliferation and in vitro vasculogenic mimicry of Ewing sarcoma cells, and to explore the possible mechanism.Methods： The expression level of miR-107 in several Ewing sarcoma cell lines was detected by real-time fluorescent quantitative PCR. The synthesized miR-107 mimic and the scramble mimic （as the control） were transfected into Ewing sarcoma RD-ES and SK-ES-1 cells, respectively. The recovery of miR-107 expression in RD-ES and SK-ES-1 cells after transfection was verified by real-time fluorescent quantitative PCR. The effects of miR-107 overexpression on the proliferation and vascular mimicry formation of RD-ES and SK-ES-1 cells were detected by CCK-8 assay and matrigel vasculogenic mimicry assay, respectively. The expressions of hypoxia inducible factor-lβ （HIF-1β） mRNA and protein in RD-ES and SK-ES-1 cells after transfection with miR-107 mimic were detected by real-time fluorescent quantitative PCR and Western blotting. Results： Comparing with human mesenchymal stem cells, miR-107 was low expressed in Ewing sarcoma RD-ES and SK-ES-1 cells （both P 〈 0.01）. After transfection with miR-107 mimic, the expression of miR-107 was significantly up-regulated in RD-ES and SK-ES-1 cells （both P 〈 0.01）. The proliferation and vascular mimicry formation of RD-ES and SK- ES-1 cells were significantly inhibited after transfection with miR-107 mimic （all P 〈 0.05）. Furthermore, miR-107 overexpression significantly suppressed the expressions of HIF-1β mRNA and protein in RD-ES and SK-ES-1 cells （all P 〈 0.01）. Conclusion： miR-107 was low-expressed in Ewing sarcoma RD-ES and SK-ES-1 cells. The overexpression of miR-107 can inhibit the proliferation and vascularization of Ewing sarcoma cells, which may be related to the targeting regulation of HIF-1β expression.

关 键 词：肉瘤 EWING 微RNA-107 低氧诱导因子1 新生血管化 病理性 细胞增殖 

分 类 号：R738.1[医药卫生—肿瘤]