siRNA阻断Stat3信号促进食管鳞癌细胞株凋亡  

Blocking Stat3 Signal by RNAi Promotes Cell Apoptosis in Esophageal Squamous Cell Carcinoma

作　　者：王新华[1] 李珊珊[1] 田芳[1] 郑献召[1] 李道明[1] 孙洋[1] 

机构地区：[1]郑州大学第一附属医院河南省肿瘤病理重点实验室,郑州450052

出　　处：《肿瘤防治研究》2010年第8期873-877,共5页Cancer Research on Prevention and Treatment

基　　金：河南省科技攻关基金资助项目(0624410048);"十五""211工程"重点学科建设基金资助项目[教重办(2002)第2号];河南省医学科技攻关基金资助项目(2006)

摘　　要：目的研究RNA干扰技术沉默Stat3基因对食管鳞癌细胞(EC9706)中持续性激活Stat3信号的阻断作用及对细胞凋亡的影响。方法将化学合成的100nM的Stat3 siRNA转染EC9706细胞,RT-PCR检测转染前后Stat3 mRNA的表达,Western blot检测转染前后细胞中Bcl-2、Stat3及磷酸化Stat3(p-Stat3)蛋白的表达,凝胶电泳迁移率(EMSA)检测转染前后活化Stat3蛋白的DNA结合活性,流式细胞术检测转染前后细胞凋亡的改变情况。结果 Stat3siRNA转染细胞48h后细胞形态发生明显的改变,RT-PCR及Western blot结果显示Stat3 siRNA干扰后Stat3 mRNA及蛋白表达均受到明显抑制,Stat3信号的激活被阻断,活化Stat3蛋白的核结合活性显著下降。转染后细胞中Bcl-2蛋白的表达明显减少。流式细胞术结果证明Stat3 siRNA可明显促进细胞凋亡。结论 Stat3 siRNA能够特异地阻断食管鳞癌细胞中Stat3信号的持续性激活并促进肿瘤细胞凋亡。Objective To investigate the blockage of constitutively activated signal transducers and activators of transcription3（Stat3） signal and the effect on cell apoptosis by applying RNA interfere technique to silence Stat3 gene in human esophageal squamous cell carcinoma cell line（EC9706）.Methods EC9706 was transfected with chemical syntheswased Stat3 siRNA（100nM）,RT-PCR and Western blotting were used to detect the expression of Stat3 mRNA,the protein levels of Bcl-2,Stat3 and phosphorylated-Stat3（p-Stat3）before and after transfection.The change of DNA-binding activity and the cells apoptosis were evaluated by electrophoretic mobility gel shift assay（EMSA）and FCM,respectively.Results After transfected Stat3 siRNA48h,cells morphology were significantly trans-formed.RT-PCR and Western blot showed the expression of Stat3 gene was inhibited at mRNA and protein levels by Stat3 SiRNA and the constitutively activated Stat3 signal was blocked.Stat3-DNA-binding activity was supresssed directly at DNA level comfirmed by EMSA.The expression of Bcl-2 protein was reduced after transfected by Stat3 siRNA.FCM indicated cell that apoptosis was promoted significantly in EC9706.Conclusion Stat3 siRNA blocked the constitutively actived Stat3 signaling specially and efficiently in EC9706,which promoted cell apoptosis obviously.

关 键 词：食管鳞癌 信号转导子及转录活化子3 RNA干扰 凋亡 

分 类 号：R735.1[医药卫生—肿瘤]