微RNA-194参与结直肠癌细胞上皮-间质转化及迁移和侵袭的作用机制探讨  

Effect of micro RNA-194 on EMT, migration and invasion of colorectal cancer cells and its potential mechanisms

作　　者：蔡洪科[1] 陈茜[2] 王继见[1] 邓甬川[3] 

机构地区：[1]重庆医科大学附属第二医院胃肠外科,重庆400010 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016 [3]浙江大学医学院附属第二医院肿瘤外科,浙江杭州310009

出　　处：《肿瘤》2016年第3期280-287,共8页Tumor

摘　　要：目的 :观察微RNA-194(micro RNA-194,mi R-194)在结直肠癌细胞侵袭和迁移过程中的作用,并探讨其参与细胞上皮-间质转化的作用机制。方法 :将携带mi R-194基因的质粒载体转染结直肠癌SW480细胞,构建稳定高表达mi R-194的细胞系mi R-194-480,并观察该细胞的形态学变化。采用MTT法、细胞划痕愈合实验和Transwell迁移及侵袭实验分别观察mi R-194-480细胞的增殖、迁移和侵袭能力;同时,采用实时荧光定量PCR和蛋白质印迹法分别检测mi R-194高表达对上皮-间质转化相关分子表达水平的影响。结果 :高表达mi R-194的结直肠癌mi R-194-480细胞系构建成功,细胞由之前的扁圆形转变为梭形,发生明显的间质性改变。mi R-194质粒转染后,mi R-194-480细胞的增殖能力并未见明显变化(P>0.05),但细胞迁移和侵袭能力却明显增强(P值均<0.001)。mi R-194高表达能够上调基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)和波形蛋白(vimentin)的表达水平(P<0.05,P<0.001),而E-钙黏蛋白(E-cadherin)表达明显下调(P<0.01)。结论 :mi R-194通过调节MMP-2、E-cadherin和vimentin的表达,参与结直肠癌细胞的上皮-间质转化,从而明显提高结直肠癌细胞的迁移和侵袭能力。Objective：To observe the effects of microRNA-194（miR-194） on migration and invasion of colorectal cancer cells,and to investigate its potential mechanisms involved in epithelial-mesenchymal transition（EMT）.Methods：The plasmids carrying miR-194 gene were transfected into colorectal cancer SW480 cells to establish a cell line named as miR-194-480 which stably and highly expressed miR-194.The cytomorphological changes of miR-194-480 cells were observed.Then the cell proliferation,migration and invasion abilities of miR-194-480 cells were detected by MTT method,wound healing assay,and Transwell migration and invasion assay,respectively.The effect of high expression of miR-194 on expressions of EMT-related proteins was detected by real-time fluorescent quantitative PCR and Western blotting,respectively.Results：The colorectal cancer cell line miR-194-480 with high expression of miR-194 was successfully constructed.As compared with the round shape of SW480 cells,the miR-194-480 cells changed to spindle-shaped,indicating visible interstitial change.After transfection with plasmids carrying miR-194 gene,the proliferation ability of SW480 cells did not change significantly（P 〉 0.05）,but the abilities of cell migration and invasion were significantly enhanced（both P 〈 0.001）.The overexpression of miR-194 up-regulated the expressions of matrix metalloproteinase-2（MMP-2） and vimentin proteins（P 〈 0.05,P 〈0.001）,but down-regulated the expression of E-cadherin（P 〈 0.01）.Conclusion：miR-194 plays an important role in EMT process of colorectal cancer cells by regulating the expressions of MMP-2,E-cadherin and vimentin,so as to enhance the abilities of migration and invasion of colorectal cancer cells.

关 键 词：结直肠肿瘤 微RNAS 肿瘤转移 肿瘤浸润 MiR-194 上皮-间质转化 

分 类 号：R735.3[医药卫生—肿瘤]