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.
作者
董希琛
汉京华
李丽娜
王雅昆
温韬
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)
出处
《中国医刊》
CAS
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）
关键词
结肠癌
T合酶
Tn抗原
上皮-间质转化
Colorectal cancer
T-synthase
Tn antigen
Epithelial-mesenchymal transition
