miR-144-3p靶向ONECUT2抑制肺癌细胞迁移、侵袭的实验研究  

Experimental study on inhibition of migration and invasion of lung cancer cells by miR-144-3p targeting ONECUT2

作　　者：鲁战胜 刘波[1] 孙勇攀[1] 孔德淼 姚静[1] 周小江 许川[1] 罗猛[1] 张庆斌[1] LU Zhansheng;LIU Bo;SUN Yongpan;KONG Demiao;YAO Jing;ZHOU Xiaojiang;XU Chuan;LUO Meng;ZHANG Qingbin(Department of Thoracic Surgery,Guizhou People s Hospital,Guiyang 550002,China)

机构地区：[1]贵州省人民医院胸外科

出　　处：《临床肿瘤学杂志》2019年第9期774-778,共5页Chinese Clinical Oncology

基　　金：贵州省基础研究计划项目(黔科合基础[2019]1202)

摘　　要：目的分析miR-144-3p对肺癌细胞迁移和侵袭的影响,并探讨其可能机制。方法采用实时荧光定量PCR(QPCR)检测肺癌NCI-H460细胞、正常支气管上皮细胞HBE中miR-144-3p、ONECUT2的表达;采用脂质体法将miR-144-3p mimics(miR-144-3p组)、miR-NC(miR-NC组)、si-con(si-con组)、si-ONECUT2(si-ONECUT2组)、miR-144-3p mimics和pcDNA(miR-144-3p+pcDNA组)、miR-144-3p mimics和pcDNA-ONECUT2(miR-144-3p+pcDNA-ONECUT2组)转染至NCI-H460细胞;Transwell法检测各组细胞的迁移、侵袭能力;Western blotting检测各组ONECUT2、E-cadherin、N-cadherin、Vimentin蛋白表达;双荧光素酶报告实验检测miR-144-3p和ONECUT2的靶向关系。结果与HBE组比较,NCI-H460组细胞中miR-144-3p表达显著降低,ONECUT2表达显著升高(P<0.05)。过表达miR-144-3p、沉默ONECUT2均可明显抑制NCI-H460细胞的迁移和侵袭能力,上调E-cadherin蛋白表达,下调N-cadherin、Vimentin蛋白表达;miR-144-3p可抑制野生型ONECUT2 3’-UTR报告基因载体的荧光素酶活性,而对突变型ONECUT2 3’-UTR-MUT的荧光素酶活性无影响。过表达ONECUT2可逆转miR-144-3p对NCI-H460细胞的EMT相关蛋白表达。结论miR-144-3p过表达可抑制肺癌细胞的迁移和侵袭能力,其机制可能与靶向ONECUT2抑制EMT有关,可为肺癌的治疗提供参考。Objective To analyze the effect of miR-144-3p on the migration and invasion of lung cancer cells and explore its possible mechanism.Real-time fluorescence quantitative polymerase chain reaction(QPCR)was used to detect the expression of miR-144-3p and ONECUT2 in lung cancer NCI-H460 cells and normal bronchial epithelial cells HBE.miR-144-3p mimics(miR-144-3p group),miR-NC(miR-NC group),si-con(si-con group),si-ONECUT2(si-ONECUT2 group),miR-144-3p mimics and pcDNA(miR-144-3p+pcDNA group),miR-144-3p mimics and pcDNA-ONECUT2(miR-144-3p+pcDNA-ONECUT2 group)were transfected into NCI-H460 cells by liposome method.Transwell assay was used to detect the migration and invasion ability of cells in each group.The protein expression of ONECUT2,E-cadherin,N-cadherin,and Vimentin of each group were detected by Western blotting.Dual luciferase reporter gene assay was used to detect the targeting relationship between miR-144-3p and ONECUT2.Results Compared with HBE group,the expression of miR-144-3p and ONECUT2 in NCI-H460 group were significantly decreased and increased(P<0.05).Overexpression of miR-144-3p or silence of ONECUT2 could significantly inhibit the migration and invasion of NCI-H460 cells,up-regulate E-cadherin protein expression,down-regulate N-cadherin and Vimentin protein expression.MiR-144-3p inhibited the luciferase activity of wild-type ONECUT2 3’-UTR reporter gene vector,but had no effect on the luciferase activity of mutant ONECUT2 3’-UTR-MUT.Overexpression of ONECUT2 reversed the expression of EMT-related protein in NCI-H460 cells by miR-144-3p.Conclusion Overexpression of miR-144-3p can inhibit the migration and invasion of lung cancer cells.The mechanism may be related to the inhibition of EMT by targeting ONECUT2,which can provide a reference for the treatment of lung cancer.

关 键 词：肺癌 miR-144-3p ONECUT2 EMT 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]