食管鳞癌放射抗拒的相关microRNA筛选  

microRNA expression profiles in radioresistant esophageal squamous carcinoma cells

作　　者：陈鑫[1] 车少敏[1] 惠蓓娜[1] 张晓智[1] 

机构地区：[1]西安交通大学医学院第一附属医院放疗科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2013年第3期296-301,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30972962)~~

摘　　要：目的探索放射抗拒对食管癌细胞microRNA(miRNA)表达的影响,筛选差异表达miRNAs,为靶向miRNAs的放射增敏研究提供理论依据。方法食管鳞癌Eca-109、TE-1细胞分别经2Gy/次的X-线照射处理,照射累积剂量达64Gy,分别命名细胞株为Eca-109R、TE-1R;克隆形成实验检测细胞放射抗拒能力;芯片检测放射抗拒细胞与母代细胞miRNA表达谱,qRT-PCR验证miRNAs表达水平。结果照射剂量40Gy后,Eca-109、TE-1细胞出现分裂受阻、多核巨型细胞、树枝状异形细胞等毒性反应,经多次传代培养完成累计剂量64Gy,恢复上皮细胞形态;0、1、2、4、6、8Gy照射后,放射抗拒细胞株Eca-109R、TE-1R增殖能力均较母代细胞增强,差异具有统计学意义(P<0.001);放射抗拒细胞株Eca-109R、TE-1R的克隆形成率中的N、Dq、D0等参数较母代细胞具有更强的放射抗拒性,差异具有统计学意义(P<0.05);miRNA芯片检测并经qRT-PCR验证,TE-1R与Eca-109R细胞较母代细胞均存在多个明显差异表达的miRNA,两放射抗拒细胞株间比较,仅miR-21在放射抗拒形成前后具有共同的表达改变趋势,且差异具有统计学意义(P<0.05)。结论放射抗拒细胞株Eca-109R、TE-1R构建成功,且与母代细胞存在差异表达的miR-NAs,放射抗拒细胞株中miR-21的表达可能与放射抗拒特性相关。Objective To explore the effects of radioresistance on microRNA（miRNA） expression of esophageal cancer cells and screen differentially expressed miRNAs to provide a theoretical basis for targeting miRNAs radiosensitization study. Methods Esophageal squamous cell carcinoma Eca-109 and TE-1 cells, treated by X-ray irradiation of 2 Gy/shoot and cumulative dose of 64 Gy respectively, were named the cell lines Eca-109R and TE-1R. We made colony formation assay of cell radioresistant capacity, chip detection of miRNA expression profile of radioresistant cells and the parent cells, and qRT-PCR validation of miRNAs expression level. Results After irradiation dose of 40 Gy, Eca-109 and TE-1 cells showed radiation toxicity like blocked split, multinucleated giant cells and heterosexual dendritic cells. A cumulative dose of 64 Gy was completed after repeated subculture; cell epithelial morphology was restored. After 0, 1, 2, 4, 6, and 8 Gy irradiation, the proliferative capacity of radioresistant cell lines Eca-109R and TE-1R was significantly greater than that of the parent ceils （P^0. 001）. N, Dq and DO of radioresistant cell lines Eca-109R and TE-1R were significantly more resistant to radiation than those of the parent cells （P^0.05）. microRNA microarray and qRT-PCR validation showed that TE-1R and Eca-109R cells had several significant differentially expressed miRNA compared with the parent cells, but only miR-21 had a common expression change before and after the formation of radioresistance in the two cell lines, with a significant difference （P〈0.05）. Conclusion and differentially expressed miRNAs radioresistant cell lines may be related Radioresistant cell lines Eca-109R were detected compared with the to the radioresistant characteristics and TE-1R were successfully constructed parent cells, miR-21 expression in the

关 键 词：MIRNA 食管癌 放射抗拒 表达谱 放射增敏 

分 类 号：R735.1[医药卫生—肿瘤]