沉默EC9706细胞NS基因对裸鼠移植瘤组织中EGF、EGFR基因表达的影响  

Silencing nucleostemin in EC9706 cells inhibits tumor growth and expression of EGF and EGFR in xenograft tumor in nude mice

作　　者：张功员[1] 关文池[2] 李晟磊[3] 张巧[1] 吴卫东[1] 

机构地区：[1]郑州大学公共卫生学院流行病学教研室,郑州450001 [2]厦门大学医学院临床医学系,福建厦门361005 [3]郑州大学基础医学院病理学教研室,郑州450001

出　　处：《第三军医大学学报》2011年第13期1370-1373,共4页Journal of Third Military Medical University

基　　金：河南省重点科技攻关基金(72102310104);河南省教育厅自然科学研究基金(2007310017);郑州市科技攻关基金(064SGDS33218-28)~~

摘　　要：目的建立裸鼠食管癌移植瘤模型,探讨由RNAi诱导的EC9706细胞核干细胞因子(nucleostemin,NS)基因沉默对裸鼠移植瘤组织中EGF、EGFR基因表达的影响。方法裸鼠15只,采用STATA 8.0编制随机化分组程序,均分为3组,分别皮下注入pRNAT-U6.1-siNS2转染的EC9706细胞(siRNA干预组),pRNAT-U6.1-siC转染的EC9706细胞(无关siRNA对照组)以及正常EC9706细胞(空白对照组),接种5周,观察移植瘤生长情况,应用免疫组织化学方法和原位杂交技术检测裸鼠移植瘤组织中NS、EGF、EGFR基因蛋白和mRNA的表达。结果无关siRNA对照组及空白对照组第4天于接种部位出现肉眼可见的肿块,siRNA干预组于第6天在相应部位发现肉眼可见肿块,第5周各组裸鼠成瘤率均为100%。siRNA干预组、无关siRNA对照组及空白对照组裸鼠移植瘤组织中NS、EGF、EGFR蛋白和mRNA的阳性表达率比较差异显著(P<0.05)。相对于无关siRNA对照组和空白对照组,siRNA干预组裸鼠肿瘤组织中NS蛋白及mRNA的阳性表达率均降低。结论 EC9706细胞沉默NS基因可降低裸鼠体内EGF、EGFR基因的表达。Objective To investigate the effect of silencing nucleostemin（NS） in EC9706 cells induced by RNAi on the expression of epidermal growth factor（EGF） and its receptor（EGFR）.Methods Fifteen nude mice were divided into three groups,each group contains five mice.Three different kinds of cells,EC9706 cells transfected by pRNAT-U6-1-siNS2,or by pRNAT-U6.1-siC and normal EC9706 cells were injected into the hypodermis tissues of corresponding mice respectively.Five weeks later after the injection,the sizes of xenograft tumors in these mice were measured,and the protein and mRNA expression of NS,EGF and EGFR in the xenograft tumor were detected by immunohistochemistry and RT-PCR.Results Visible tumors were observed in the injected position after 4 d in transfected pRNAT-U6.1-siC and normal EC9706 cells groups,and after 6 d in transfected pRNAT-U6-1-siNS2 EC9706 cells group.In 5th week,the tumorigenesis were 100%.The protein and mRNA levels of NS,EGF and EGFR were lower in transfected pRNAT-U6-1-siNS2 EC9706 cells group than in the other two groups（P〈0.05）.Conclusion Silencing NS of EC9706 cells inhibits the growth of exnograft tumor in nude mice and decreases the expression of EGF gene and EGFR gene.Silencing NS gene may be a new strategy for the treatment of esophagus caicinoma.

关 键 词：食管鳞状细胞癌 EC9706细胞 RNA干扰 裸鼠 核干细胞因子 

分 类 号：R394.3[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]