miR-124靶向STAT3改善脑恶性胶质瘤细胞辐射敏感性  

miR-124 increases the radiosensitivity of glioblastoma cells through targeting STAT3

作　　者：马建[1] 王新军[1] 付旭东[1] 周少龙[1] 田碧[2] 闫琳[3] Ma Jian;Wang Xinjun;Fu Xudong;Zhou Shaolong;Tian Bi;Yan Lin(Department of Neurosurgery,The Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450000, China;Department of Radiotherapy,The Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Oncology,The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450000,China)

机构地区：[1]郑州大学第五附属医院神经外科,450000 [2]郑州大学第五附属医院放疗科,450000 [3]郑州大学第五附属医院肿瘤科,450000

出　　处：《中华放射医学与防护杂志》2019年第2期88-94,共7页Chinese Journal of Radiological Medicine and Protection

摘　　要：目的研究miR-124在放射敏感及耐受的脑恶性胶质瘤细胞LN229和LN229R中的表达以及miR-124对细胞放射敏感性的影响,并深入探讨miR-124调控LN229R细胞放射敏感性的作用机制。方法将miR-124模拟物(miR-124)及阴性对照(miR-NC),STAT3过表达质粒(STAT3)及pcDNA3.1质粒(pcDNA)单独或共转染到放射抵抗的胶质瘤细胞LN229R中。qRT-PCR检测LN229和LN229R细胞中miR-124的表达;克隆形成实验分析不同放射剂量下LN229R细胞的生存率以及敏感性相关参数值;流式细胞术分析LN229R细胞的凋亡情况;生物信息学预测miR-124和STAT3的靶向关系,并利用双荧光素酶报告分析加以验证;Westernblot分析STAT3的蛋白表达水平。结果与LN229组(1.02±0.09)相比,miR-124在LN229R细胞中的表达水平(0.32±0.03)显著降低,差异有统计学意义(t=12.780,P<0.05);与miR-NC组(0.95±0.06)相比,转染miR-124模拟物可促进LN229R细胞中miR-124的表达(4.02±0.39)(t=13.476,P<0.05);8Gy照射条件下,miR-124过表达组癌细胞的生存率(0.003±0.0004)显著低于miR-NC组(0.033±0.0050)(t=5.655,P<0.05),细胞凋亡率(22.34±2.42)%显著高于miR-NC组(4.69±0.51)%(t=12.361,P<0.05);STAT3是miR-124的靶基因;外源回补STAT3可逆转miR-124对LN229R细胞生存的抑制作用。结论miR-124通过靶向STAT3改善LN229R细胞的放射敏感性。Objective To investigate the expression of miR-124 in glioblastoma (GBM) cell lines LN229 and LN229R, as well as the regulatory mechanism of miR-124 on radiosensitivity of LN229R cells. Methods miR-124 mimic (miR-124) and negative control (miR-NC), STAT3 overexpression plasmid (STAT3) and pcDNA3.1 vector (pcDNA) were transfected or co-transfected into radioresistant glioma cells LN229R. qRT-PCR was employed to analyze the expression of miR-124 in LN229 and LN229R cells. The survival rate and sensitivity-related parameters of LN229R cells at different doses were analyzed by cloning formation assay. Cell apoptosis of LN229R was evaluated by flow cytometry. Targeting gene of miR-124 was predicted using Targetscan software and verified by the double-luciferase reporter assay. Western blot assay was performed to detect STAT3 protein expression. Results The expression of miR-124 in LN229R cells (0.32±0.03) was significantly lower than that in LN229 cells (1.02±0.09) (t=12.780, P<0.05). Transfection of miR-124 mimics promoted the expression of miR-124 in LN229R cells (4.02±0.39) compared with miR-NC group (0.95±0.06) (t=13.476, P<0.05). After 8 Gy irradiation, the survival rate of LN229R cells transfected with miR-124 mimics (0.003±0.000 4) was significantly lower than that in miR-NC group (0.033±0.005 0) (t=5.655, P<0.05), and the apoptosis rate (22.34±2.42)% was significantly higher than that in miR-NC group (4.69±0.51)% (t=12.361, P<0.05). STAT3 was identified to be a target gene of miR-124. Exogenous restoration of STAT3 reversed the inhibitory effect of miR-124 on LN229R cell survival. Conclusion miR-124 increases the radiosensitivity of LN229R cells by targeting STAT3.

关 键 词：胶质瘤细胞 放射敏感性 miR-124 STAT3 

分 类 号：R739.4[医药卫生—肿瘤] R730.55[医药卫生—临床医学]