食管癌细胞系中Nanog基因沉默对顺铂敏感性的影响  

The Effect of Silencing Nanog Gene Expression on Sensitization to Cisplatin in Esophageal Cancer Cell-Line

作　　者：石磊芝[1] 杜亚明[1] 苏鹏 刘长军 王中彬[1] 

机构地区：[1]辽宁医学院附属第一医院,121000 [2]辽宁省辽阳市中心医院胸外科 [3]辽宁省鞍山第二医院

出　　处：《天津医药》2012年第4期312-315,I0001,共5页Tianjin Medical Journal

基　　金：辽宁省科学技术计划项目(项目编号:2011225015);辽宁医学院青年科技启动基金项目(项目编号:Y2011Z001)

摘　　要：目的:检测干细胞基因Nanog在食管癌细胞系TE-1中的表达及其对顺铂敏感性的影响。方法:通过细胞免疫荧光检测食管癌细胞系TE-1中Nanog蛋白的表达;应用Lipofectamine2000将荧光染料标记的Nanog siRNA转染进细胞中,通过荧光显微镜观察转染效率,再通过逆转录聚合酶链反应(RT-PCR)和Western blot检测小干扰RNA(siRNA)的沉默效率;转染后24h加入顺铂,继续孵育48h后测得不同转染组的半数抑制浓度(IC50),后加入相同浓度的顺铂5mg/L,48h后测得不同转染组的生存率。结果:Nanog蛋白在食管癌细胞系TE-1中高表达,以核表达为主;Nanog siRNA细胞转染率为(67.57±16.90)%,沉默效率可高达85%以上;转染24h后加入顺铂,继续孵育48h后测得Nanog siRNA组的IC50较阴性对照组和空白组相比均降低,差异有统计学意义。加入相同浓度顺铂5mg/L,48h后测得Nanog siRNA组细胞生存率明显低于阴性对照组和空白对照组,差异有统计学意义。结论:Nanog在食管癌细胞系TE-1中高表达,降低Nanog的表达后能够提高食管癌细胞对顺铂的敏感性。Objective： To investigate effects of expression of stem cell gene Nanog on esophageal cancer cell （ESCC） line TE-1 and on the sensitization to cisplatin. Methods：The expression of Nanog in ESCC cell line TE-1 was detected by immunofluoreseence. The fluorescence labeled small interfering RNA （siRNA） was transfected into ceils using Lipofectamine 2000. The transfection efficiency was observed by fluorescence microscopy. The silencing efficiency of siRNA was evaluated by RT-PCR and Western blot. Cisplatin was added 24 hours after transfection. After 48 h cultivation, the ICso was measured in different transfection groups. The survival rates were detected in different transfection groups after adding cisplatin 5mg/L for 48 h. Results： The expression of Nanog protein was higher in ESCC cell line TE-1, and the expression was mainly in the nucleus. The cell transfection rate was （67.57± 16.90）%. Silencing efficiency of Nanog siRNA was reached to 85% compared with that in negative control group. The value of IC50 was significantly decreased in Nanog siRNA group than that of negative control group and blank group. The cell survival rate was significantly decreased in Nanog siRNA group after treatment with 5 mg/L cisplatin for 48 h compared with that of negative control group and blank group. Conclusion ： Nanog is highly expressed in ESCC cell line TE-1. Nanog siRNA efficiently reduces Nanog expression, which enhances cisplatin chemosensitivity in ES- CC cell line.

关 键 词：食管肿瘤 癌 细胞系 肿瘤RNA 小分子干扰 顺铂 抗药性 肿瘤 逆转录聚合酶链反应 印迹法 蛋白质Nanog 

分 类 号：R735.1[医药卫生—肿瘤]