miR-155在非小细胞肺癌组织和细胞中的表达及对细胞增殖、迁移、侵袭的影响

The expression of miR-155 in NSCLC tissues and cells and its effects on cell proliferation,migration and invasion

目的:探讨miR-155在非小细胞肺癌(NSCLC)中的表达及其对NSCLC细胞增殖、迁移、侵袭的影响机制。方法:通过生物信息学网站预测miR-155的可能靶基因,双荧光素酶报告基因实验进行验证;采用qRT-PCR测定NSCLC组织及细胞中miR-155和ZIC3的表达;通过Lipofectamine^(TM)2000试剂盒向A549细胞中分别转染miR-155 mimic和mimic NC,共转染miR-155 mimic+pMIR-ZIC3;Western blot检测转染miR-155 mimic对ZIC3蛋白表达的影响;MTT法、划痕实验及Transwell侵袭实验分别检测miR-155对A549细胞增殖、迁移及侵袭的影响。结果:生物信息学网站预测显示ZIC3-3'UTR与miR-155存在结合位点,双荧光素酶报告基因实验验证ZIC3是miR-155的靶基因,ZIC3表达受miR-155的负向调控。NSCLC组织及细胞中miR-155显著低表达,ZIC3显著高表达(P<0.05);过表达miR-155抑制了ZIC3蛋白的表达水平(P<0.05)。转染miR-155 mimic显著抑制NSCLC细胞的增殖、迁移及侵袭能力(P<0.01);共转染miR-155 mimic+pMIR-ZIC3逆转了miR-155 mimic对NSCLC细胞增殖、迁移及侵袭的抑制作用(P<0.05)。结论:NSCLC中miR-155显著低表达,其过表达可抑制NSCLC细胞的增殖、迁移及侵袭;miR-155可能通过靶向负调控ZIC3影响NSCLC细胞的生物学行为。

Objective:To investigate the expression of miR-155 in non-small cell lung cancer(NSCLC)and the mechanism of its influence on the proliferation,migration and invasion of NSCLC cells.Methods:The possible target genes of miR-155 were predicted through bioinformatics sites and verified by double luciferase reporter gene experiment.The expressions of miR-155 and ZIC3 in NSCLC tissues and cells were determined by qRT-PCR.Lipofectamine^(TM)2000 kit was used to transfect miR-155 mimics and NC mimics into A549 cells,and miR-155 mimics+pMIR-ZIC3 were co-transfected.Western blot was used to detect the effects of transfection with miR-155 mimic on the protein expression of ZIC3.The effects of miR-155 on the proliferation,migration and invasion of A549 cells were determined by MTT assay,scratch assay and Transwell invasion assay,respectively.Results:The bioinformatics site predicted that ZIC3-3'UTR and miR-155 had binding sites.The dual-luciferase reporter gene experiment verified that ZIC3 was the target gene of miR-155,and ZIC3 expression was negatively regulated by miR-155.miR-155 expression was significantly lower and ZIC3 expression was significantly higher in NSCLC tissues and cells(P<0.05).The overexpression of miR-155 inhibited the expression of ZIC3 protein(P<0.05).Transfection of miR-155 mimic significantly inhibited the proliferation,migration and invasion of NSCLC cells(P<0.01).Co-transfection of miR-155 mimic+pMIR-ZIC3 reversed the inhibitory effects of miR-155 mimic on proliferation,migration and invasion of NSCLC cells(P<0.05).Conclusion:The expression of miR-155 in NSCLC was significantly lower,and its overexpression could inhibit the proliferation,migration and invasion of NSCLC cells.miR-155 may inhibit the biological behavior of NSCLC cells by targeting negative regulation of ZIC3 expression.