小干扰RNA阻断信号转导子和转录激活子3信号对人食管鳞癌细胞株增殖的抑制作用  

Small interfering RNA suppression of transducer and activator of transcription 3(STAT3)signaling pathway:inhibitory effect on proliferation of human esophageal squamous carcinoma cells

作　　者：王新华 李珊珊[1] 阎爱华[1] 孙洋 卢创新 郭燕萍 

机构地区：[1]郑州大学第一附属医院病理科河南省肿瘤病理重点实验实 郑州大学基础医学院病理学教研室,450052

出　　处：《中华病理学杂志》2007年第6期379-383,共5页Chinese Journal of Pathology

基　　金：河南省科技攻关基金(0424410003);国家"十五"攻关;教育部"211工程"重点学科建设基金[教重办(2002)第2号]

摘　　要：目的探讨信号转导子和转灵激活子3(STAT3)小干扰RNA(siRNA)对食管鳞状细胞癌细胞株(EC9706和Eca109)中持续性激活STAT3信号的阻断情况及阻断STAT3信号对细胞增殖的影响。方法将化学合成的100 nmol/L的STAT3 siRNA转染EC9706和Eca109细胞,逆转录聚合酶链反应检测转染前后STAT3 mRNA的表达,Western blot检测转染前后STAT3及磷酸化STAT3 (p-STAT3)蛋白的表达,凝胶电泳迁移率检测转染前后活化STAT3蛋白的核结合情况,力甲基偶氮唑蓝(MTT)检测转染前后细胞增殖能力的改变,流式细胞仪检测转染前后细胞周期的改变。结果STAT3 siRNA以时间依赖方式特异性地抑制STAT3 mRNA及STAT3、p-STAT3蛋白的表达,并且STAT3蛋白的核结合活性下降,使细胞周期阻滞于G_0/G_1期,细胞增殖受到明显的抑制。与对照组相比EC9706细胞转染后72 h G_0/G_1期的细胞增加了16.1%,同时S期细胞减少11.1%;Eca109细胞转染后72 h G_0/G_1期的细胞增加了11.8%,同时S期细胞也较对照组明显减少。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抑制肿瘤细胞的增殖。Objective To investigate the efficiency of blockage of constitutively activated STAT3 signaling by small interfering RNA （siRNA）, and to explore the inhibitory effects on the proliferation of human esophageal squamous carcinoma cells （EC9706 and Eca109）. Methods EC9706 and Eca109 were transfected with chemical synthesized STAT3 siRNA（ 100 nmol/L）. RT-PCR and Western blot were used to detect STAT3 mRNA and protein expression, including phosphorylated-STAT3 （p-STAT3） before and after the transfection respectively. The changes of DNA-binding activity and cell proliferation were evaluated by electrophoretic mobility gel shift assay and MTT, respectively. Stages of cell cycle were determined by flow cytometry. Results Expression levels of STAT3 mRNA and STAT3, p-STAT3 proteins were progressively inhibited by STAT3 siRNA at various time points after transfection. STAT3-DNA-binding activity was suppressed after transfection evidenced by electrophoretic mobility gel shift assay. The cell cycle was arrested at G0/G1 phase along with a significant inhibition of cell proliferation after STAT3 siRNA treatment. Conclusion STAT3 siRNA specifically and efficiently blocks the constitutively activated STAT3 signaling pathway in human esophageal squamous carcinoma cells, resulting in cell cycle arrest and proliferation inhibition.

关 键 词：食管肿瘤 癌 鳞状细胞 STAT3转录因子 RNA干扰 细胞系 肿瘤 

分 类 号：R735.1[医药卫生—肿瘤]