MiR-25对三阴乳腺癌侵袭转移的影响及其潜在机制  

The effects of miR-25 and the potential molecular mechanisms on migration and invasion of triple negative breast cancer

作　　者：吴唐维[1] 蒋丽媛 张天竺 郑超 刘水逸[1] 李晓怡 陈卫群[1] 卢忠心[1] Wu Tangwei;Jiang Liyuan;Zhang Tianzhu;Zheng Chao;Liu Shuiyi;Li Xiaoyi;Chen Weiqun;Lu Zhongxin(Department of Medical Laboratory,the Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China)

机构地区：[1]华中科技大学同济医学院附属武汉中心医院检验科,武汉430014

出　　处：《中华检验医学杂志》2019年第2期104-111,共8页Chinese Journal of Laboratory Medicine

基　　金：湖北省自然科学基金重点项目(2015CFA078);武汉市卫生计生委资助项目(WX14B10,WX15A12,WX18Y11).

摘　　要：目的探讨微小RNA-25(miR-25)在三阴乳腺癌患者血浆、组织和细胞系中的表达情况及其对三阴乳腺癌侵袭转移影响的潜在分子机制。方法横断面研究。运用实时荧光定量PCR检测86例三阴乳腺癌患者和38名正常对照者血浆中miR-25的水平;应用LinkedOmics网站分析平台分析三阴乳腺癌与非三阴乳腺癌样本中miR-25的水平;同时用定量PCR检测三阴乳腺癌细胞系中miR-25的表达情况;运用划痕和Transwell实验检测miR-25抑制剂或对照转染后,三阴乳腺癌细胞系迁移和侵袭能力的变化;应用荧光素酶报告基因技术验证1-磷酸鞘氨醇磷酸化酶SGPP1是否为miR-25的靶基因;采用SGPP1过表达质粒和miR-25过表达质粒共同转染MDA-MB-231细胞,运用划痕和Transwell实验检测其对细胞迁移和侵袭能力的影响;免疫印迹Westernblot检测SGPP1蛋白水平的变化。正态分布的组间样本均数比较用Student′st检验。结果86例三阴乳腺癌患者血浆中miR-25的水平明显高于正常对照组(P为0.031);LinkedOmics网站分析平台分析显示,三阴乳腺癌标本中的miR-25表达水平明显高于常见的非三阴乳腺癌LuminalA型和LuminalB型(P分别为<0.001和0.006);在三阴乳腺癌细胞系HS578T、HCC1806、MDA-MB-231和BT549中miR-25的表达均明显高于乳腺上皮细胞系HBL-100(P分别为0.006、0.01、0.029和0.046);划痕实验显示,在MDA-MB-231和HS578T细胞中转染miR-25抑制剂后,较对照组相比,伤痕愈合率受到明显抑制(P分别为0.035和0.001);Transwell实验表明,转染miR-25抑制剂的MDA-MB-231和HS578T的细胞侵袭能力明显低于阴性对照组(P分别为0.002和0.001);LinkedOmics平台分析显示,三阴乳腺癌患者癌组织中miR-25与SGPP1表达呈负相关(P为0.037);双荧光素酶报告基因证实,SGPP1是miR-25的直接靶基因;抑制miR-25能增加SGPP1的蛋白水平;共转染SGPP1过表达质粒和miR-25过表达质粒能明显削弱miR-25过表达引起的细胞迁移和侵袭能力�Objective To explore the expression of tiny RNA-25(microRNA-25,miR-25)in the plasma、tissues of triple-negative breast cancer(TNBC)patients and cell lines,to investigate the potential molecular mechanisms of miR-25 on migration and invasion of TNBC.MethodsReal-time fluorescent quantitative PCR was used to detect the expression of miR-25 in the plasma of TNBC patients.Linked omics web platform was used to analyse miR-25 level in samples of TNBC and non-TNBC.Real-time fluorescent quantitative PCR was also used to detect the miR-25 level in TNBC cell lines.The wound healing and transwell assay was applied to assess the effects on migration and invasion of TNBC cell lines which transfected with miR-25 inhibitor or the negative control.The luciferase reporter assay was used to validate the relationship between miR-25 and the sphingosine-1-phosphate phosphatase 1(SGPP1)in HEK293T cell.The wound healing and transwell assay was used to detect the migration and invasion ability of TNBC cell lines when cotransfected with pCMV6-SGPP1 and miR-25.Furthermore,Western blot was performed to detect the SGPP1 level in TNBC cell lines.ResultsThe expression of miR-25 was significantly elevated in the plasma of 86 TNBC patients compared with the healthy controls(P value was 0.031).LinkedOmics web platform analysis showed that miR-25 expression was significantly higher in TNBC samples than in non-TNBC samples with Luminal A or Luminal B(P value was<0.001 and 0.006).The level of miR-25 was also elevated in TNBC cell lines HS578T,HCC1806,MDA-MB-231 and BT549(P value was 0.006,0.01,0.029 and 0.046).The MDA-MB-231 and HS578T cells which transfected with miR-25 inhibitor exhibited a significant slower wound healing rate than control(P value was 0.035 and 0.001).At the same time,when transfected with miR-25 inhibitor,MDA-MB-231 and HS578T both exhibited a decreased invasion ability compared with the control group(P value was 0.002 and 0.001).LinkedOmics web platform analysis showed that sphingosine-1-phosphate phosphatase 1(SGPP1)gene leve

关 键 词：三阴性乳腺癌 微RNAS 膜蛋白质类 磷酸单酯水解酶类 肿瘤转移 

分 类 号：R737.9[医药卫生—肿瘤]