肿瘤相关糖类抗原Tn异常表达促进人结肠癌细胞的恶性程度研究  

Tumor-associated carbohydrate antigen-Tn promotes malignance in human colorectal cancer cells

作　　者：董希琛 汉京华 李丽娜 王雅昆 温韬[1] DONG Xi-chen;HAN Jing-hua;LI Li-na;WANG Ya-kun;WEN Tao(Medical Research Center,Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China;120 Emergency Center,Shandong Rizhao 276800,China)

机构地区：[1]首都医科大学附属北京朝阳医院医学研究中心,北京100020 [2]山东省日照市120紧急救援指挥中心,山东日照276800

出　　处：《中国医刊》2020年第2期171-174,共4页Chinese Journal of Medicine

基　　金：国家自然基金面上项目（81672338）

摘　　要：目的探讨肿瘤相关糖类抗原Tn异常表达对人结肠癌细胞SW480的生物学行为的影响及可能的作用机制。方法采用CRISPR/Cas9技术靶向敲除人结肠癌细胞SW480的T合酶基因以诱导Tn抗原异常表达,应用流式细胞术鉴定Tn抗原的表达情况;利用Transwell小室法检测Tn抗原表达对SW480细胞迁移和侵袭能力的影响;最后采用蛋白质印迹法检测Tn异常表达对细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白E-钙黏蛋白、N-钙黏蛋白、波形蛋白的影响。结果蛋白质印迹法结果显示CRISPR/Cas9成功敲除了SW480细胞的T合酶基因,并导致Tn抗原异常表达。Tn抗原表达明显促进SW480细胞的迁移和侵袭能力。此外,Tn抗原表达显著激活了EMT信号通路,表现为上皮相关蛋白E-钙黏蛋白表达水平下调和间质相关蛋白N-钙黏蛋白和波形蛋白表达水平上调。结论肿瘤相关糖类抗原Tn异常表达可促进结肠癌细胞的迁移和侵袭,其机制可能激活EMT信号通路有关。Objective To investigate the effect of tumor-related carbohydrate antigen-Tn on oncogenic feature in human colorectal cancer cells(migration,invasion)and to explore the possible mechanism.Method Using CRISPR/Cas9 system to knockout the gene encoding T-synthase in a CRC cell line SW480.Expression of Tn antigen and deletion of T-synthase were determined by flow cytometry and western blotting.Migration and invasion were performed by Transwell chamber assay and Matrigel.The effects of Tn expression on the epithelialmesenchymal transition(EMT)-related proteins in SW480 were detected by western blotting.Result CRISPR/Cas9 system knocked out T-synthase entirely and knockout of T-synthase induced Tn antigen expression in SW480 cells.Expression of Tn antigen induced by T-synthase deficiency promotes the cell migration and invasion of SW480 cells.Expression of Tn antigen down-regulates the expression level of EMT-related protein E-cadherin,up-regulates the expression level of N-cadherin and Vimentin in SW480 cells.Conclusion Expression of Tn antigen induced by T-synthase deficiency enhances the migration and invasion of colorectal cancer SW480 cells via activation of EMT.

关 键 词：结肠癌 T合酶 Tn抗原 上皮-间质转化 

分 类 号：R735.3[医药卫生—肿瘤]