MiR-192对结直肠癌细胞ZEB2表达及迁移和侵袭的影响  

Effects of miR-192 on the expression of ZEB2 and the abilities of migration and invasion of colorectal cancer cells

作　　者：吴巍芸[1] 梁坚[1] 叶石才[1] 周宇[1] 

机构地区：[1]广东医学院附属医院消化内科,广东湛江524001

出　　处：《肿瘤》2014年第1期13-18,共6页Tumor

摘　　要：目的:探讨miR-192对结直肠癌SW480和SW620细胞ZEB2(zinc-finger E-box binding homeobox2)表达及迁移和侵袭能力的影响。方法:将miR-192模拟物(miR-192 mimics)分别转染SW480和SW620细胞,应用实时荧光定量RT-PCR和蛋白质印迹法检测转染后SW480和SW620细胞中miR-192、ZEB2 mRNA和ZEB2蛋白的表达,细胞划痕实验和Transwell侵袭实验检测转染后SW480和SW620细胞迁移和侵袭能力的改变。将重组载体pmiR-ZEB2-wt和miR-192 mimics共转染入SW480细胞,应用双荧光素酶报告基因验证ZEB2基因3'端非翻译区(3'-untranslated region,3'-UTR)是否有miR-192的结合位点。结果:与转染miR-192模拟物阴性对照组比较,miR-192 mimics转染后SW480和SW620细胞中miR-192的表达水平上调(P<0.05),ZEB2mRNA及其蛋白的表达水平明显下调(P<0.05),细胞的迁移和侵袭能力下降(P<0.05)。双荧光素酶报告基因验证结果显示,pmiR-ZEB2-wt和miR-192 mimics共转染后,SW480细胞中荧光素酶活性下降(P<0.05),ZEB2基因3'-UTR存在miR-192的结合位点。结论:ZEB2可能是miR-192的靶基因之一,上调SW480和SW620细胞中miR-192的表达可抑制ZEB2的表达,发挥抑制细胞迁移和侵袭的作用。Objective： To investigate the effects of miR-192 on the expression of zinc-finger E-box binding homeobox 2 （ZEB2） and the abilities of migration and invasion of colorectal cancer （CRC） SW480 and SW620 cells. Methods： After transfection of miR-192 mimics into SW480 and SW620 cells, the expression levels of miR-192 and ZEB2 mRNA and ZEB2 protein were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively. The abilities of migration and invasion of SW480 and SW620 cells after transfection with miR-192 mimics were determined by wound healing and Transwell assays, respectively. The recombinant vector pmiR-ZEB2-wt and miR-192 mimics were co- transfected into SW480 cells. The binding site of 3＇-untranslated region （3＇-UTR） of ZEB2 gene with miR-192 was verified by Dual LuciferaseTM Reporter Gene Assay. Results： As compared with the negative control （NC） group （transfection with miR-192 mimics-NC）, the expression levels of miR-192 in SW480 and SW620 cells after transfection with miR-192 mimics were increased （P 〈 0.05）, and the expression levels of ZEB2 mRNA and protein were decreased （P 〈 0.05）, as well as the abilities of migration and invasion of SW480 and SW620 cells were inhibited （P 〈 0.05）. The Dual LuciferaseTM Reporter Gene Assay revealed that the luciferase activity in SW480 cells after co-transfection with recombinant vector pmiR- ZEB2-wt and miR-192 mimics was inhibited （P 〈 0.05）, which indicated that the 3＇-UTR of ZEB2 harbored a binding site for miR-192. Conclusion： ZEB2 may be one of the target genes for miR-192. Overexpression of miR-192 may down-regulate the ZEB2 expression and inhibit the migration and invasion of SW480 and SW620 cells.

关 键 词：结直肠肿瘤 微RNAs肿瘤侵润 ZEB2 细胞 SW480 SW620 

分 类 号：R735.3[医药卫生—肿瘤]