微小RNA-21反义寡核苷酸对食管癌顺铂耐药细胞的影响  

Impact of microRNA-21 antisense oligonucleotide on cisplatin resistance of EC9706/DDP cells

作　　者：潘雪[1] 李向楠[2] 

机构地区：[1]郑州大学护理学院,450052 [2]郑州大学第一附属医院胸外科

出　　处：《中华实验外科杂志》2015年第5期1072-1074,共3页Chinese Journal of Experimental Surgery

基　　金：河南省卫生厅医学科技攻关计划资助项目（2011020029）

摘　　要：目的 观察微小RNA-21（miRNA-21）反义寡核苷酸对食管癌EC9706顺铂（DDP）耐药细胞（EC9706/DDP）的影响.方法 设计miRNA-21反义寡核苷酸,建立EC9706/DDP耐药细胞株并转染耐药细胞,实时荧光定量聚合酶链反应（FQ-PCR）检测各组细胞miRNA-21表达水平,噻唑蓝（MTT）法检测各组细胞对DDP药物敏感性、克隆形成实验检测各组细胞增殖能力,流式细胞术检测细胞凋亡变化,Western blot检测B细胞淋巴瘤/白血病-2（bcl-2）、B细胞淋巴瘤/白血病-2相关X蛋白（bax）蛋白表达.结果 与空白组、miRNA-阴性对照（NC）比较,miRNA-21反义寡核苷酸组miRNA-21相对表达量下降为0.21 ±0.89、半数抑制剂量（IC50）明显降低为（16.22 ±1.85） mg/L、克隆集落的数量较少、细胞凋亡率明显增高为（30.5±2.9）％、bcl-2蛋白表达水平降低,bax蛋白表达水平增高,差异均有统计学意义（P＜0.05）.结论 miRNA-21可能通过降低bcl-2、增高bax蛋白表达水平降低EC9706/DDP细胞对DDP的耐药性.Objective To investigate the impact of microRNA （miRNA）-21 antisense oligonucleotide on cisplatin （DDP） resistance of EC9706/DDP cells.Methods miRNA-21 antisense oligonucleotides were designed and transfected into EC9706/DDP-resistant cells.The miRNA-21 expression levels were detected by real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）.Methyl thiazol tetrazolium （MTT） method was used to measure the sensitivity of cells to DDP drugs.Colony formation assay was used to test the cell proliferation.The change of apoptosis was examined by flow cytometry.Western blotting was performed to detect the expression of B cell lymphoma/leukemia-2 （bcl-2）,and bcl-2 associated X protein （bax） proteins.Results As compared with miRNA-NC negative control group and blank control group,the miRNA-21 relative expression was decreased to （0.21 ±0.89）,50% inhibitory dose （IC50） decreased to （16.22 ± 1.85） mg/L,the number of clonal colonies was reduced,the apoptosis rate significantly increased to （30.5 ± 2.9） %,bcl-2 protein expression decreased,and bax protein expression increased in miRNA-21 antisense oligonucleotide group （P 〈 0.05）.Conclusion miRNA-21 decreased the resistance of EC9706/DDP cells to DDP possibly through reducing the bcl-2 protein expression,and increasing the bax protein expression.

关 键 词：微小RNA-21 食管癌 顺铂 

分 类 号：R735.1[医药卫生—肿瘤]