ACSS2介导的上皮间质转化在乙醇促结直肠癌细胞迁移中的作用及机制  

Role and mechanism of ACSS2-mediated epithelial-mesenchymal transition in ethanol-promoted migration of colorectal cancer

作　　者：孙燕 王灵巧[2] 张国伟[2] 孙凤琼 王文钰 徐桂勇 张爱华[1] 周紫垣[2] 杨光红[1] SUN Yan;WANG Lingqiao;ZHANG Guowei;SUN Fengqiong;WANG Wenyu;XU Guiyong;ZHANG Aihua;ZHOU Ziyuan;YANG Guanghong(School of Public Health,Guizhou Medical University,Guiyang,Guizhou Province,550025,China;Department of Environmental Health,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,400038,China)

机构地区：[1]贵州医科大学公共卫生与健康学院,贵阳550025 [2]陆军军医大学(第三军医大学)军事预防医学系环境卫生学教研室,重庆400038

出　　处：《陆军军医大学学报》2023年第17期1806-1818,共13页Journal of Army Medical University

基　　金：国家自然科学基金面上项目(81273156)。

摘　　要：目的 探索ACSS2(acyl-CoA synthetase short chain family member 2)在乙醇促结直肠癌(colorectal cancer, CRC)细胞迁移过程中的作用及其潜在机制。方法 以在线工具基因表达谱交互式分析(gene expression profiling interactive analysis, GEPIA)和人类蛋白质图谱数据库(the human protein atlas, HPA)分析CRC癌组织和癌旁正常组织中ACSS2表达水平;通过ChIP-X富集分析3转录因子数据库(ChIP-X enrichment analysis 3,ChEA3)进行ACSS2上游转录因子的富集分析。从癌症基因组图谱(the cancer genome Atlas, TCGA)数据库获得513例CRC患者的ACSS2表达水平与相关临床特征参数。以不同浓度乙醇(0、22、44、88 mmol/L)处理HCT116、SW480细胞24、48、72 h, CCK-8实验检测细胞活力;细胞划痕和Transwell实验检测细胞迁移能力;RT-qPCR和Western blot检测相关基因的表达。结果 (1)以22、44、88 mmol/L浓度的乙醇处理48、72 h能显著抑制HCT116和SW480细胞增殖(P<0.05)。(2)乙醇处理48 h后,RT-qPCR和Western blot结果显示HCT116、SW480细胞中ACSS2的表达明显增加;细胞划痕和Transwell结果显示细胞愈合率和穿孔细胞数均明显增加(P<0.05);上皮间质转化(epithelial-mesenchymal transformation, EMT)相关蛋白E-cadherin表达降低、N-cadherin和Vimentin表达升高(P<0.05)。(3)抑制ACSS2表达后,细胞愈合率和穿孔细胞数均较未抑制组降低(P<0.05);E-cadherin蛋白表达增加、而N-cadherin和Vimentin蛋白表达降低(P<0.05)。(4)HPA数据库分析提示,ACSS2在CRC癌组织中的表达明显高于正常组织,其上游调控因子的富集分析提示有8个转录因子可能与ACSS2的表达调控有关,Western blot验证乙醇处理后转录因子CEBPB、SMC1A、CTCF在SW480和HCT116细胞中的表达均显著上调,与ACSS2的表达增加变化一致。(5)513例CRC患者中,ACSS2相对高表达组出现外周神经、血管浸润或发生其他远处转移者的比例明显高于相对低表达组(χ2=6.411,P=0.011)。结论 乙醇作用后由�Objective To explore the role and underlying mechanisms of Acyl-CoA synthetase short chain family member 2(ACSS2)in ethanol-promoted migration of colorectal cancer(CRC)cells.Methods The ACSS2 expression level in cancer tissues and adjacent normal tissues was analyzed by online tools Gene Expression Profiling Interactive Analysis(GEPIA)and Human Protein Atlas(HPA).The transcription factor enrichment analysis tool ChIP-X Enrichment Analysis 3(ChEA3)was used to identify ACSS2 upstream transcription factors.The ACSS2 expression level in cancer tissue and related clinical characteristic parameters of 513 CRC patients were obtained from The Cancer Genome Atlas(TCGA)database.After HCT116 and SW480 cells were treated with ethanol at a dose of 0,22,44 and 88 mmol/L for 24,48 and 72 h,respectively,cell viability,migration and expression of related molecules at mRNA and protein levels were detected with CCK-8 assay,cell scratch test and transwell assay,and RT-qPCR and Western blotting respectively.Results(1)After treated by ethanol for 48 and 72 h,the 3 doses of 22,44 and 88 mmol/L showed a significant inhibition of proliferation for both HCT116 and SW480 cells(P0.05).(2)After 48 h of ethanol treatment,RT-qPCR and Western blotting revealed that the expression levels of ACSS2 were increased significantly in HCT116 and SW480 cells,cell scratch test and Transwell assay indicated that the cell healing rate and number of perforated cells were also increased obviously(P0.05),and epithelial-mesenchymal transformation(EMT)related proteins,E-cadherin was down-regulated,while N-cadherin and Vimentin were up-regulated(all P0.05).(3)Inhibition of ACSS2 expression resulted in obviously decreased cell healing rate and number of perforated cells(P0.05),enhanced expression level of E-cadherin and decrased levels of N-cadherin and Vimentin(all P0.05).(4)HPA database analysis showed that ACSS2 has a significantly higher expression level in CRC cancer tissues than in normal tissues,and upstream regulators enrichment analysis suggested 8 trans

关 键 词：乙醇 ACSS2 结直肠癌 迁移 上皮间质转化 

分 类 号：R730.23[医药卫生—肿瘤] R735.35[医药卫生—临床医学] R595.6