Shh信号通路对结肠癌细胞EMT作用的研究  

Effect of Shh signaling pathway on EMT in colon cancer cells

作　　者：孙亚超[1] 王静东[2] 雷程[1] 孙振强[1] 王海江[1] 王琦三[1] SUN Yachao;WANG Jingdong;LEI Cheng;SUN Zhenqiang;WANG Haijiang;WANG Qisan(Department of Gastrointestinal Surgery,the Affiliated Tumor Hospital,Xinjiang Medical University,Urumqi,Xinjiang,830011,China;~General Surgery,Xinjiang Armed Police general Hospital,Urumqi,Xinjiang,830052,China)

机构地区：[1]新疆医科大学附属肿瘤医院胃肠外科,乌鲁木齐830011 [2]武警新疆总队医院普外科,乌鲁木齐830052

出　　处：《新疆医学》2018年第5期492-495,共4页Xinjiang Medical Journal

基　　金：新疆维吾尔自治区自然科学基金(2013211A066)

摘　　要：目的观察Shh信号通路活化及阻断后结肠癌细胞形态及上皮间质表型转化(Epithelial-Mesenchymal Transition,EMT)相关标志物上皮型钙黏着蛋白(E-cadherin)和波形蛋白(Vimentin)表达的改变,进一步探讨Shh信号通路对结肠癌细胞上皮间质表型转化(EMT)的作用。方法通过常规培养结肠癌细胞株HT-29,设为三组,分别是:培养液中加入PBS(对照组)、培养液中加入重组Shh配体(信号通路活化组)、培养液中加入Shh信号通路抑制剂KADD-cyclopamine(信号通路阻断组)。通过应用RT-PCR、Westen-blot试验来检测上皮间质转化标志物E-cadherin和Vimentin在m RNA水平及蛋白表达的变化。结果RT-PCR试验显示:与对照组相比较,活化组的Vimentin的m RNA表达量明显升高(P<0.05)、E-cadherin的表达量明显降低(P<0.05),信号活化组与阻断组之间的两种蛋白表达有差异且具有统计学意义(P<0.05);Westen-blot试验检测蛋白显示:激活Shh信号通路后,活化组结肠癌细胞E-cadherin蛋白表达减弱,蛋白vimentin表达增强,而阻断组结肠癌细胞E-cadherin蛋白表达增强,蛋白vimentin表达减弱,且两组相比较有统计学意义(P<0.05)。结论激活Shh信号通路能够使结肠癌细胞发生上皮间质转化,抑制其活化可以降低结肠癌细胞的转变作用,结肠癌的转移可能与EMT的发生密切相关。Objective To explore the role of Shh signaling pathway on epithelial mesenchymal transition（EMT） in colon cancer cells by observing the changes of the expression of E-cadherin and vimentin after Shh signaling pathway activation and blockade.Methods Colon cancer HT-29 cells were divided into control group（PBS being added into medium）, activatingsignaling pathways group（recombinant shh ligand was added into medium）and blocking signaling pathways group（Shh inhibitor KADD-cyclopamine was added into culture medium）. After 24 h of transfection, cultured cells were harvested to detect the changes of E-cadherin and Vimentin protein by RT-PCR and Western-blot respectively.Results The results of RT-PCR showed that the expression of epithelial cell marker E-cadherin was decreased and the m RNA expression of vimentin was enhanced at the activating Shh signaling pathway group compared with those of control group, and there was a significant difference in the expression levels of E-cadherin and Vimentin between the activated group and blocked group（P〈0.05） at the m RNA level. Westen-blot results showed that the expression of E-cadherin was decreased and expression of vimentin was increased in colon cancer cells after Shh signaling pathway was activated, while the expression of E-cadherin was increased and expression of vimentin was decreased in the blocking group. There was a significant difference between the activated group and blocked group（P〈0.05）. Conclusion Activating the Shh signaling pathway can promote EMT occurrence and development of colon cancer cells, while the inhibition of its activation can reduce the transformation of colon cancer cells, so cancer metastasis may be closely related with EMT occurrence.

关 键 词：SHH信号通路 结肠癌细胞 上皮间质表型转化 

分 类 号：R735.35[医药卫生—肿瘤]