乙醇通过促进上皮间质转化增强结直肠癌细胞的侵袭转移能力  

Ethanol promotes migration and invasion in colorectal cancer cells via epithelial-mesenchymal transition

作　　者：夏宜馨 张国伟[2] 王灵巧[2] 龙琦 陈维艳 谭瑶[2] 杨建英 向梦龙[2] 周紫垣[2] 张爱华 XIA Yixin;ZHANG Guowei;WANG Lingqiao;LONG Qi;CHEN Weiyan;TAN Yao;YANG Jianying;XIANG Menglong;ZHOU Ziyuan;ZHANG Aihua(School of Public Health,Guizhou Medical University,Guiyang,Guizhou Province,550025;Department of Military Enviionment Health,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,400038;Guiyang Maternal and Child Health Care Hospital,Guiyang,Guizhou Province,550025,China)

机构地区：[1]贵州医科大学公共卫生学院,贵阳550025 [2]陆军军医大学(第三军医大学)军事预防医学系军事环境卫生学教研室,重庆400038 [3]贵阳市妇幼保健院,贵阳550025

出　　处：《第三军医大学学报》2020年第15期1501-1510,共10页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81273156)。

摘　　要：目的研究乙醇对结直肠癌细胞HT29、SW480增殖、侵袭迁移能力的影响,并探讨其作用机制。方法利用CCK-8实验检测乙醇对HT29和SW480细胞增殖活性的影响,采用相关分析得出IC50值,建立结直肠癌细胞株低浓度乙醇染毒模型;流式细胞术检测细胞周期分布,细胞划痕实验、Transwell小室实验检测细胞迁移、侵袭能力,流式细胞术检测细胞ROS水平,Western blot检测细胞E-cadherin、N-cadherin和vimentin等上皮间质转化(epithelial-mesenchymal transition,EMT)相关蛋白表达情况,利用乙醛检测试剂盒检测乙醇作用下乙醛的浓度。结果乙醇对HT29、SW480细胞的IC50值分别为436、424 mmol/L;随着乙醇浓度的增大,处于G1期细胞的占比增大;细胞划痕实验结果显示,与对照组相比,各浓度组乙醇处理后,HT29、SW480细胞愈合百分比皆明显增加(P<0.05);Transwell实验显示,在无基质胶和有基质胶条件下,乙醇处理皆能显著增加HT29、SW480细胞穿透小室的细胞数量(P<0.05)。进一步实验结果显示,乙醇能明显增加HT29、SW480细胞内的ROS水平(P<0.05);Western blot实验结果显示乙醇作用后HT29、SW480细胞内EMT相关蛋白E-cadherin表达显著降低,N-cadherin和vimentin的表达明显增加(P<0.05);乙醇浓度的升高,细胞中乙醛浓度随之增大。结论乙醇可能通过ROS介导EMT途径增强结直肠癌细胞的侵袭迁移能力。Objective To determine the effects of ethanol on the cell proliferation,migration and invasion in human colorenctal cancer HT29 and SW480 cells and investigate the underlying mechanisms.Methods The effect of ethanol on the proliferation of HT29 and SW480 cells was detected by CCK-8 assay.After the IC50 value was obtained by the relevant analysis,and low concentration ethanol exposure models(1/10 IC50 value)were established.The cell cycle distribution was measured by flow cytometry,cell migration and invasion were detected by cell scratch test and Transwell chamber assay respectively,and the cell ROS production were detected using flow cytometry.The protein levels of epithelial-mesenchymal transition(EMT)related proteins,such as E-cadherin,N-cadherin and vimentin were detected by Western blotting.The production of acetaldehyde in the cells after ethanol treatment was detected with acetaldehyde detection kit.Results The IC50 values of ethanol to HT29 and SW480 cells were 436 and 424 mmol/L,respectively.With the increase of ethanol concentration,the proportion of cells at G1 stage was increased.Compared with the control group,the cell scratch test showed that the percentages of wound closure of HT29 and SW480 cells were increased significantly after ethanol treatment in each concentration group(P<0.05).Transwell chamber test indicated that ethanol treatment significantly increased the number of HT29 and SW480 cells permeating without and with matrigel(P<0.05).Further experiments showed that ethanol significantly enhanced the intracellular ROS level of HT29 and SW480 cells(P<0.05).Western blotting displayed that ethanol decreased the expression level of E-cadherin,while increased those of N-cadherin and vimentin(P<0.05).And as the concentration of ethanol increasing,the acetaldehyde content was elevated in the cells.Conclusion Ethanol may enhance the invasion and migration of colorectal cancer cells through ROS-mediated EMT pathway.

关 键 词：乙醇 结直肠癌 迁移 侵袭 上皮间质转化 

分 类 号：O623.411[理学—有机化学] R730.23[理学—化学]