LncRNA FoxP4-AS1靶向miR-136-5p影响人乳腺癌细胞株SKBR-3侵袭与迁移能力  

LncRNA FoxP4-AS1 targeting miR-136-5p affects the invasion and migration ability of human breast cancer cell line SKBR-3

作　　者：孙传伟 蒋曼妃 吴煌福 王璐瑶 谭光宏[1] SUN Chuan-Wei;JIANG Man-Fei;WU Huang-Fu(Department of Breast and Thyroid Surgery,the Second Affiliated Hospital of Hainan Medical University,Haikou 570100,Hainan,China)

机构地区：[1]海南医学院第二附属医院乳甲外科,海南海口570100 [2]海南医学院第二附属医院全科医学科,海南海口570100

出　　处：《中国老年学杂志》2023年第19期4746-4751,共6页Chinese Journal of Gerontology

基　　金：2018年度国家自然科学基金委员会项目(81860650);海南省卫生健康行业科研项目(22A200070)。

摘　　要：目的探讨长链非编码核糖核酸(LncRNA)FoxP4-AS1靶向微小核糖核酸(miR)-136-5p对人乳腺癌细胞株SKBR-3侵袭与迁移能力的作用。方法将处于对数生长期的人乳腺癌细胞株SKBR-3分为FoxP4-AS1下调组(转染si-FoxP4-AS1)、FoxP4-AS1上调组(转染pcDNA-FoxP4-AS1)、FoxP4-AS1对照组(转染FoxP4-AS1-NC)、miR-136-5p下调组(转染miR-136-5p inhibitor)、miR-136-5p上调组(转染miR-136-5p mimic)、miR对照组(转染miR-NC)和空白组(不转染)。Transwell测定细胞侵袭、迁移能力;实时逆转录定量聚合酶链反应(RT-qPCR)检测FoxP4-AS1、miR-136-5p表达及迁移侵袭增强因子(MIEN)1、E-钙黏蛋白(E-cadherin)、基质金属蛋白酶(MMP)-9信使核糖核酸(mRNA)表达;Western印迹检测MIEN1、E-cadherin、MMP-9蛋白表达;双荧光素酶报告基因实验验证FoxP4-AS1靶向调控miR-136-5p。结果与空白组和FoxP4-AS1对照组比,FoxP4-AS1上调组侵袭和迁移细胞数目明显增多,miR-136-5p表达、E-cadherin mRNA及蛋白表达明显降低,FoxP4-AS1表达、MIEN1、MMP-9 mRNA及蛋白表达明显升高(P<0.05)。与空白组和miR对照组比,miR-136-5p上调组侵袭和迁移细胞数目明显减少,miR-136-5p、E-cadherin表达明显增高,MIEN1、MMP-9表达明显下降(P<0.05)。双荧光素酶报告基因实验结果证实FoxP4-AS1靶向调控miR-136-5p。结论FoxP4-AS1可促进乳腺癌细胞株SKBR-3侵袭与迁移能力,可能是通过靶向调控miR-136-5p的表达,促进MIEN1、MMP-9的表达,抑制E-cadherin的表达实现该作用的。Objective To investigate the effect of long chain non encoding ribonucleic acid(LncRNA)FoxP4-AS1 targeting microRNA(miR)-136-5p on invasion and migration of human breast cancer cell line SKBR-3.Methods Human breast cancer cell line SKBR-3 at logarithmic growth stage was divided into FoxP4-AS1 down-regulated group(transfected with si-FoxP4-AS1),FoxP4-AS1 up-regulated group(transfected with pcDNA-FoxP4-AS1),FoxP4-AS1 control group(transfected with FoxP4-AS1-NC),miR-136-5p down-regulated group(transfected with miR-136-5p inhibitor),miR-136-5p up-regulated group(transfected with miR-136-5p mimic),miR control group(transfected with miR-NC)and blank group(non-transfection).Transwell was used to detect the ability of cell invasion and migration.Real time reverse transcription quantitative polymerase chain reaction(RT-qPCR)was used to detect the expressions of FoxP4-AS1,miR-136-5p,migration and invasion enhancing factor(MIEN)1,E-cadherin and matrix metalloproteinase(MMP)-9 messenger RNA(mRNA).The protein expressions of MIEN1,E-cadherin and MMP-9 were detected by Western blot.Double luciferase reporter gene experiment verified that FoxP4-AS1 targeted regulation of miR-136-5p.Results Compared with the blank group and FoxP4-AS1 control group,the number of invasive and migratory cells was significantly increased,the expressions of miR-136-5p and E-cadherin mRNA and protein were significantly decreased,while the expressions of FoxP4-AS1,MIEN1,MMP-9 mRNA and protein were significantly increased in FoxP4-AS1 up-regulated group(P<0.05).Compared with the blank group and miR control group,the number of invasive and migratory cells was significantly decreased,the expressions of miR-136-5p and E-cadherin were significantly increased,and the expressions of MIEN1,MMP-9 were significantly decreased in miR-136-5p up-regulated group(P<0.05).The results of double luciferase reporter gene experiment confirmed that FoxP4-AS1 targeted regulation of miR-136-5p.Conclusions FoxP4-AS1 could promote the invasion and migration of breast cancer cel

关 键 词：长链非编码核糖核酸FoxP4-AS1 微小核糖核酸-136-5p 乳腺癌 侵袭 迁移 

分 类 号：R737.9[医药卫生—肿瘤]