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辐射诱导性miR-34a的表达对细胞辐射敏感性的影响
程赢,和生辉,刘聪,周传丰,崔建国,孙顶,蔡建明,李百龙,高福*
0
(第二军医大学海军医学系放射医学教研室,上海 200433)
摘要:
目的探讨miR-34a的表达与细胞、组织辐射敏感性的关系。方法采用RT-PCR检测辐射前后不同组织、细胞中miR-34a的表达量;利用流式细胞术检测细胞凋亡率,MTT法检测细胞活力;在肿瘤细胞U87MG中转染miR-34a mimics上调miR-34a的表达并检测其细胞活力;在正常细胞HEK293中转染miR-34a inhibitors下调miR-34a的表达水平并检测其细胞凋亡率。结果照前辐射敏感性高的细胞miR-34a表达水平高,辐射敏感性低的细胞miR-34a表达水平低;照后辐射敏感性高的细胞miR-34a表达上调幅度远高于辐射敏感性低的细胞。上调miR-34a表达水平使得肿瘤细胞U87MG照后细胞活力降低,下调miR-34a表达使得正常细胞HEK293照后细胞凋亡率降低。结论细胞辐射敏感性与miR-34a的表达水平正相关,辐射敏感性高的细胞照后miR-34a的表达上调幅度更大;上调miR-34a表达水平对肿瘤细胞U87MG有明显的辐射增敏作用,下调miR-34a表达对正常细胞HEK293有明显的辐射防护作用。
关键词:  miR-34a  辐射耐受性  辐射增敏药  辐射防护
DOI:10.3724/SP.J.1008.2011.0368
投稿时间:2011-04-06修订日期:2011-04-11
基金项目:国家自然科学基金(30970679,31070761).
Effect of irradiation-induced miR-34a expression on irradiation-sensitivity of cells
CHENG Ying,HE Sheng-hui,LIU Cong,ZHOU Chuan-feng,CUI Jian-guo,SUN Ding,CAI Jian-ming,LI Bai-long,GAO Fu*
(Department of Radiation Medicine, Faculty of Navy Medicine, Second Military Medical University, Shanghai 200433, China)
Abstract:
ObjectiveTo investigate the relationship between miR-34a expression and the irradiation-sensitivity of cells and tissues.MethodsRT-PCR was used to detect the miR-34a expression in different cells and tissues before and after irradiation; flow cytometry was employed to examine the apoptosis of cells and MTT assay was used to observe the cell viability. The U87MG tumor cells were transfected with miR-34a mimics to promote miR-34a expression and the cell viability was observed 24 h later. The HEK293 renal epithelial cells were transfected with miR-34a inhibitors to decrease miR-34a expression and the cells apoptosis was observed 24 h later.ResultsThe expression of miR-34a was higher in highly irradiation-sensitive cells than in lowly irradiation-sensitive cells before irradiation; the increase of miR-34a expression was greatly higher in highly irradiation-sensitive cells than in lowly irradiation-sensitive cells 24 h after irradiation. The up-regulation of miR-34a decreased the viability of U87MG tumor cells, and down-regulation of miR-34a decreased the apoptosis of HEK293 cells.ConclusionThe irradiation-sensitivity of cells is positively associated with miR-34a; cells with high irradiation-sensitivity have a greater up-regulation of miR-34a expression after irradiation. Increased miR-34a expression can noticeably increase the irradiation-sensitivity of U87MG tumor cells and decreased miR-34a expression has a irradiation prevention effect on normal HEK293 cells.
Key words:  miR-34a  radiation tolerance  radiation-sensitizing agents  radiation protection