激活Shh信号通路影响结肠癌细胞上皮间质表型转化的研究  

The effect of activating Shh signaling pathway on epithelial-mesenchymal transition of colon cancer cells activating

作　　者：孙亚超[1] 王静东[2] 金博[1] 孙振强[1] 方法[1] 王琦三[1] SUN Yachao WANG Jingdong JIN Bo SUN Zhenqiang FANG Fa WANG Qisan(Department of Gastrointestinal Surgery, Affiliated Tumor Hospital, Xinjiang Medical University, Urumqi 830011, China Department of General Surgery, Xinjiang Uyger Autonomous Region Corps Hospital, Chinese People＇s Armed Poloce Force, Urumqi 830096, China)

机构地区：[1]新疆医科大学附属肿瘤医院腹外科,乌鲁木齐830011 [2]武警新疆总队医院普外科,乌鲁木齐830096

出　　处：《新疆医科大学学报》2016年第11期1428-1431,共4页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2013211A066)

摘　　要：目的阐释激活Shh信号通路对结肠癌上皮间质表型转化(EMT)的影响。方法常规培养结肠癌细胞株HT-29细胞,设立对照组(培养液中加入PBS)、实验组(即信号通路活化,培养液中加入重组Shh配体);采用Westen-blot检测E-cadherin和Vimentin蛋白表达水平的变化;RT-PCR检测上皮间质转化标志物E-cadherin和Vimentin在mRNA水平的变化;行HE染色观察细胞形态变化的改变。结果 Westen-blot检测蛋白提示,结肠癌HT-29细胞中,Shh信号通路激活可促进Vimentin蛋白表达(P<0.05),抑制E-cadherin蛋白表达(P<0.05);RT-PCR检测:实验组中E-cadherin和Vimentin的mRNA表达量差异具有统计学意义(P<0.05),与对照组相比较,实验组的Vimentin的mRNA表达量显著升高(P<0.05),E-cadherin的表达量明显降低(P<0.05);HE细胞染色提示对照细胞呈梭形,细胞间更紧密地连接,并与信号通路阻断组细胞无显著差异,而实验组细胞呈椭圆形或圆形,连接疏松,部分细胞呈团簇集群样生长。结论 EMT在结肠癌转移过程中起着重要的作用,而Shh信号通路的激活可促进结肠癌细胞EMT的发生、发展。Objective To explain effect of Shh signaling pathway on epithelial-mesenchymal transition（ EMT） of colon cancer cells. Methods Routinely cultured colon cancer HT-29 cells were devided intocontrol group（ PBS in broth） and testing group（ recombinant shh ligand in PBS）; 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. The morphological changes of the cells were observed by HE staining. Res-ults Westen-blot analysis showed that activation of Shh signaling pathway can promote the expression of Vimentin（ P〈 0. 05） and inhibit the E-cadherin protein expression（ P〈 0. 05） in colon cancer HT-29 cells; Compared with control group,the mRNA expression of Vimentin significantly increased（ P〈 0. 05） and the expression of E-cadherin significantly decreased（ P 〈0. 05） in the experimental group; HE staining showed that the cell of control group were fusiform,closely connected,and had no significant difference with the signal path blocking group,however,the signaling pathway activated cells were round or oval,loose connection,and some were clustered. Conclusion EMT plays an important role in colon cancer metastasis,and activating the Shh signaling pathway can promote the EMT of colon cancer cell.

关 键 词：上皮间质表型转化 SHH信号通路 结肠癌 E-cadherin和Vimentin 

分 类 号：R34[医药卫生—基础医学]