结肠癌中上皮细胞黏附分子与Wnt-β-连环蛋白信号传导之间的关系及XAV939对相关标志物表达的影响  

Relationship between epithelial cell adhesion molecule and wingless/integrated-β-catenin signaling in colon cancer and the effect of XAV939 on the expression of related markers

作　　者：刘磊[1] 周逢强[1] 齐艳美[2] 王海艳 耿振 张禄[1] 宋峰峰 丁宝忠[1] 郭辉光[1] 徐宏[1] 李乐平[4] Liu Lei;Zhou Fengqiang;Qi Yanmei;Wang Haiyan;Geng Zhen;Zhang Lu;Song Fengfeng;Ding Baozhong;Guo Huiguang;Xu Hong;Li Leping(Department of Gastrointestinal Surgery,Binzhou People’s Hospital,Binzhou 256600,China;Department of Digestive Surgery,Binzhou People’s Hospital,Binzhou 256600,China;Department of General Foreign Affairs Binzhou People’s Hospital,Binzhou 256600,China;Department of General Surgery,Shandong University Affiliated Provincial Hospital,Binzhou 256600,China)

机构地区：[1]滨州市人民医院胃肠外科,256600 [2]滨州市人民医院消化外科,256600 [3]滨州市人民医院普外三科,256600 [4]山东大学附属省立医院普外科,滨州256600

出　　处：《中华实验外科杂志》2019年第9期1601-1604,共4页Chinese Journal of Experimental Surgery

基　　金：山东省医药卫生科技发展计划项目(2017WSA16030).

摘　　要：目的观察在结肠癌中Wnt-β-连环蛋白(β-catenin)通路的激活是否调节上皮细胞黏附分子(EpCAM)表达,并分析XAV939对结肠癌相关标志物表达的影响.方法激活Wnt-β-catenin信号传导后,通过荧光实时定量聚合酶链反应(RT-qPCR)确定相关基因相对表达水平.构建小鼠结肠癌模型,腹腔注射XAV939和/或5-Fu处理1周后,通过蛋白印迹法检测结肠组织中相关分子表达.应用SPSS 24.0统计软件分析,以独立样本t检验进行组间比较,以配对t检验进行组内比较.结果与对照组比较,LiCl能够显著激动Wnt-β-catenin信号传导通路相关基因的表达,TACSTD1、DKK1、BAMBⅠ、CCND1和MYC基因水平均显著升高,分别上升1.20、2.44、4.34、2.57、1.58倍(t=12.179、20.661、54.378、19.384、20.341,P<0.05).ZnCl2处理则能显著抑制HCT116细胞中TACSTD1和CCND1的表达(tTACSTD1=36.467、9.682,tCCND1=16.130、17.003,P<O.05),分别下调50.06%、34.38%.而在APC基因缺失细胞中,ZnCl2对TACSTD1和CCND1的抑制作用被逆转.XAV939和XAV939+5-氟尿嘧啶(5-Fu)处理能够显著上调HCT116细胞中Axin相对表达水平,较对照组的升高15.97倍和17.20倍(t=19.502、18.236,P<0.05);同时β-catenin、EpCAM、端粒反转录酶(TERT)和双皮质素和钙/钙调蛋白依赖性蛋白激酶样-1(DCAMKL-1)的表达水平分别下调0.75和0.72倍、0.68和0.25倍、0.74和0.73倍、0.27和0.34倍(tβ-catenin=20.412、22.862,tEpCAM=26.336、27.111、82.711,tTERT=19.636、22.787、23.678,tDCAMKL-1=81.637、64.814,P<0.05).结论XAV939通过阻断Wnt-β-catenin信号传导下调β-catenin、靶基因EpCAM和CSC其他相关因子表达以抑制肿瘤细胞增殖,以促进结肠癌预后.Objective To investigate whether activation of Wingless/integrated-β-catenin(Wnt-β-catenin)pathway regulates epithelial cell adhesion molecule(EpCAM)expression in colon cancer and analyze the effect of XAV939 on colon cancer-associated markers.Methods After Wnt-β-catenin signaling was activated,relative expression levels of related genes were determined by quantitative real-time polymerase chain reaction(RT-qPCR).After constructing a mouse colon cancer model,XAV939 and/or 5-Fu were intraperitoneally injected for 1 week.Detection of related molecules expression in colon tissue by Western blotting.Results Compared with the control group,LiCl could significantly stimulate the expression of Wnt-β-catenin signaling pathway genes,and the levels of TACSTD1,DKK1,BAMBⅠ,CCND1 and MYC genes were significantly increased by 1.20-,2.44-,4.34-,2.57-,1.58 fold(t=12.179,20.661,54.378,19.384,20.341,P<0.05),respectively.ZnCl2 treatment significantly inhibited the expression of TACSTD1 and CCND1 in HCT116 cells(tTACSTD1=36.467,9.682,tCCND1=16.130,17.003,P<0.05),which were down-regulated by about 50.06%and 34.38%,respectively.In the APC gene-deficient cells,the inhibitory effect of ZnCl2 on TACSTD1 and CCND1 was reversed.XAV939 and XAV939+5-fluorouracil(5-Fu)treatment significantly up-regulated the expression of Axin in HCT116 cells,which was about 15.97 and 17.2 fold higher than that of the control group(t=19.502,18.236,P<0.05);and expression ofβ-catenin,EpCAM,telomerase reverse transcriptase(TERT)and bicorticosteroids and calmodulin dependent protein kinase-like 1(DCAMKL-1)were down-regulated by 0.75 and 0.72 fold,0.68 and 0.25 fold,0.74 and 0.73 fold,0.27 and 0.34 fold(tβ-catenin=20.412,22.862,tEpCAM=26.336,27.111,82.711,tTERT=19.636,22.787,23.678,tDCAMKL-1=81.637,64.814,P<0.05),respectively.Conclusion XAV939 inhibits tumor cell proliferation by blocking Wnt-β-catenin signaling down-regulation ofβ-catenin,target gene EpCAM and other related factors of CSC to promote colon cancer prognosis.

关 键 词：Wnt-β-连环蛋白通路 上皮细胞黏附因子 XAV939 结肠癌 

分 类 号：R735.35[医药卫生—肿瘤]