下调Ets2表达对裸鼠食管鳞癌移植瘤生长及mTOR/p70S6K信号通路的影响  

Effect of downregulating Ets2 on tumor growth and mTOR/p70S6K signaling pathway in nude mice xenografted esophageal squamous cell carcinoma

作　　者：韩康[1] 马珊珊[1] 李庆华[1] 王欣欣[2] 朱相展 孟楠[2] 韩莉[2] 关方霞[1,2] 

机构地区：[1]郑州大学生命科学学院,郑州450001 [2]郑州大学第一附属医院,郑州450052

出　　处：《郑州大学学报（医学版）》2016年第6期690-694,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目81471306;U1404313;河南省高等学校重点科研项目15A310028

摘　　要：目的:探讨下调Ets2表达对裸鼠Eca109移植瘤生长及mTOR/p70S6K信号通路的影响。方法:选择已筛选出的在体外靶向下调Ets2基因表达的siRNA片段,构建包含此片段的慢病毒(LV-siE ts2)并感染Eca109,15只裸鼠随机分为空白对照组、阴性对照组和LV-siE ts2组,分别背部皮下接种Eca109、LV感染的Eca109和LV-siE ts2感染的Eca109,形成移植瘤,之后观察肿瘤生长情况、绘制肿瘤生长曲线并计算肿瘤抑制率。应用qRT-PCR检测肿瘤组织中Ets2 mRNA的表达,TUNEL法检测肿瘤组织细胞的凋亡,Western blot法检测肿瘤组织中Ets2、Caspase-3、Bcl-2、E-cadherin以及mTOR/p70S6K信号通路蛋白的表达。结果:LV-siE ts2组肿瘤生长速度明显变慢,肿瘤体积和质量显著小于空白对照组和阴性对照组(P<0.05);LV-siE ts2组肿瘤组织中Ets2 mRNA及蛋白表达水平均显著降低;LV-siE ts2能引起Eca109细胞的凋亡,且上调Caspase-3和E-cadherin蛋白表达水平,而下调Bcl-2蛋白表达水平(P<0.05);LV-siE ts2能显著降低mTOR及p70S6K的蛋白表达。结论:LV-siE ts2在体内通过抑制mTOR/p70S6K信号通路促进细胞凋亡,从而抑制裸鼠Eca109移植瘤的生长。Aim： To observe the effect of downregulating expression of Ets2 on the tumor growth and mTOR/p70S6K signaling pathway in nude mice xenografted Eca109 tumor. Methods： The siRNA which could down-regulate the expression of Ets2 effectively in vitro had been selected before. LV containing the siRNA（ LV-siE ts2） was established and infected Eca109. Fifteen nude mice were randomly allocated into control group,negative control group and experimental group.Eca109,Eca109 infected by LV and Eca109 infected by LV-siE ts2 were inoculated subcutaneously into nude mice to establish transplantation tumor. Later,we observed tumor growth,drawn the tumor growth curve and calculated the tumor inhibition rate. The expression of Ets2 mRNA in tumor tissue was detected by qRT-PCR and cell apoptosis was detected by TUNEL. The protein expression of Ets2,Caspase-3,Bcl-2,E-cadherin and mTOR/p70S6K signaling pathway in tumor tissue was detected by Western blot. Results： The growth rate of tumors in the experimental group slowed down significantly and tumor size and weight was less than the control group and negative control group（ P〈0. 05）,besides,the mRNA and protein expression of Ets2 in tumor tissue from the experimental group was reduced significantly. The results of TUNEL demonstrated that LV-siE ts2 could significantly induce apoptosis of Eca109 cells（ P〈0. 05）. The protein expression levels of Caspase-3 and E-cadherin increased and the protein expression levels of Bcl-2 decreased. In addition,The results of Western blot demonstrated that LV-siE ts2 could down-regulate the protein expression level of mT OR and p70S6 K significantly. Conclusion： LV-siE ts2 could promote cell apoptosis by inhibiting mTOR/p70S6K signaling pathway in vivo,thus inhibiting tumor growth in nude mice xenografted Eca109 tumor.

关 键 词：Ets2 SIRNA ECA109 裸鼠 凋亡 mTOR/p70S6K信号通路 

分 类 号：R735.1[医药卫生—肿瘤]