论著 Original Article

同期放化疗诱导人结直肠癌细胞发生上皮-间质转化

Published at: 2017年第37卷第1期

蒋永新 2 , 王艳俊 1 , 刘珊 2 , 王红 1
1 昆明医科大学第三附属医院肿瘤研究所,昆明 650118
2 昆明医科大学第三附属医院云南省中西医结合肿瘤临床研究中心,昆明 650118
通讯作者 永新 蒋 Email: 598104374@qq.com
DOI: 10.3978/j.issn.2095-6959.2017.01.020
基金:
云南省卫生厅项目 2014NS037
云南省教育厅基础研究项目 2016ZDX061
昆明医科大学硕士研究生创新基金项目 2016S35

摘要

目的:探讨放化疗抵抗的结直肠癌细胞发生上皮–间质转化(epithelial-mesenchymal transition,EMT)的意义。方法:采用5-FU化疗同期进行放疗对人结直肠癌野生型细胞(HCT116)进行干预,诱导放化疗共同抵抗的细胞株(HCT116CRR)并采用克隆形成实验进行放化疗抵抗性的鉴定。高倍显微镜下观察细胞形态学变化。采用Real-time PCR和Western印迹,检测上皮表型标志物E-cadherin,间质表型标志物N-cadherin、波形蛋白(vimentin)、核转录因子(Snail)mRNA及其蛋白的表达。结果:放化疗抵抗的结直肠癌细胞发生与EMT相符的形态学改变,细胞呈纺锤体状,极性消失,并出现伪足;Real-time PCR和Western印迹结果显示E-cadherin mRNA及蛋白表达下调;N-cadherin,vimentin,Snail mRNA及蛋白表达上调,差异有统计学意义(P<0.05)。结论:放化疗抵抗后的人结直肠癌细胞发生EMT,其与结直肠癌的治疗抵抗相关。


Chemoradiotherapy can induce epithelial-mesenchymal transition of the human colorectal cancer cells

Abstract

Objective: To investigate the significance of epithelial-mesenchymal transition (EMT) of human colorectal cancer cells with resistance to chemoradiotherapy. Methods: Colorectal cancer cells were exposed to 5-fluorouracil (5-FU) while radiotherapy was given at the same time. A small number of cells that survived from chemoradiotherapy were obtained, and their morphological changes were observed by microscopy. Our group then identified the resistance using colony formation assay. Real-time PCR and Western blot were used to detect the mRNA and its protein expression of epithelial marker E-cadherin, mesenchymal marker N-cadherin, vimentin, and nuclear transcription factor snail. Results: Colorectal cancer cells which were resistant to chemoradiotherapy showed phenotypic changes consistent with EMT: spindle-cell shape, loss of polarity, and pseudopodia formation. Real-time PCR and Western blot results: E-cadherin mRNA and protein expression level were lower, and N-cadherin, Snail, vimentin mRNAs and proteins expression level were higher, the difference was statistically significant (P<0.05). Conclusion: Colorectal cancer cells resistance to chemoradiotherapy can induce epithelial-mesenchymal transition (EMT), and EMT may be related to resistance of chemoradiotherapy of colorectal cancer.


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引用

引用本文: 永新 蒋, 艳俊 王, 珊 刘, 红 王. 同期放化疗诱导人结直肠癌细胞发生上皮-间质转化[J]. 临床与病理杂志, 2017, 37(1): 115-120.
Cite this article as: JIANG Yongxin, WANG Yanjun, LIU Shan, WANG Hong . Chemoradiotherapy can induce epithelial-mesenchymal transition of the human colorectal cancer cells[J]. Journal of Clinical and Pathological Research, 2017, 37(1): 115-120.