摘要
目的 探讨长链非编码RNA(lncRNA)尿路上皮癌相关基因1(UCA1)在乳腺癌细胞中的表达及其对乳腺癌细胞增殖、迁移和侵袭的影响.方法 采用定量逆转录聚合酶链反应(qRT-PCR)检测UCA1在正常乳腺上皮细胞株MCF-10A和乳腺癌细胞株MCF-7、SK-BR-3、MDA-MB-231中的表达.通过小干扰RNA(siRNA)构建UCA1低表达细胞株,质粒构建UCA1过表达细胞株,采用四甲基偶氮唑蓝(MTT)法、平板克隆法检测细胞增殖能力,qRT-PCR和蛋白质印迹(Western blot)法检测细胞中细胞周期蛋白D1(cyclin D1)基因和蛋白表达量以及基质金属蛋白酶2(MMP2)蛋白表达情况,细胞划痕和Transwell实验检测细胞的迁移和侵袭能力.结果 与正常乳腺上皮细胞株MCF-10A相比,UCA1在乳腺癌细胞株中的表达均升高,以MCF-7细胞株最高;敲低UCA1可抑制MCF-7细胞的增殖、集落形成、细胞划痕愈合、迁移和侵袭(P﹤0.05),且cyclin D1基因和蛋白水平以及MMP2蛋白表达量均下调.过表达UCA1可促进MCF-7细胞的增殖、集落形成、细胞划痕愈合、迁移和侵袭(P﹤0.05),且cyclin D1基因和蛋白水平以及MMP2蛋白表达量均上调.结论 UCA1在乳腺癌细胞中表达升高,可能是通过调控cyclin D1和MMP2促进乳腺癌的增殖、迁移和侵袭.
Objective To investigate the expression of long non-coding RNA(lncRNA)urothelial cancer associated 1(UCA1)in breast cancer cells and its effect on breast cancer cell proliferation,migration,and invasion.Method Quan-titative reverse transcription-polymerase chain reaction(qRT-PCR)was used to detect the expression of UCA1 in normal breast epithelial cell line MCF-10A and breast cancer cell lines MCF-7,SK-BR-3,and MDA-MB-231.Construction of UCA1 low-expression cell lines by small interfering RNA(siRNA).UCA1 overexpressed cell lines was constructed using plasmids.The methyl thiazolyl terazolium(MTT)method and plate cloning method were used to detect cell proliferation,qRT-PCR and Western blot method were used to detect cyclin D1 gene and protein expression and matrix metalloprotein-ase 2(MMP2)protein expression in cells.Cell scratching and Transwell experiments were used to detect the migration and invasion of cells.Result Compared with normal breast epithelial cell line MCF-10A,the expressions of UCA1 in breast cancer cell lines were increased,and MCF-7 cell line was the most significant.UCA1 knockdown could inhibit MCF-7 cell proliferation,colony formation,cell scratch healing,migration,and invasion(P<0.05),the levels of the cyclin D1 gene and protein and the expression of the MMP2 protein were all down-regulated.UCA1 overexpression could pro-mote proliferation,colony formation,cell scratch healing,migration,and invasion of MCF-7 cells(P<0.05),the levels of cyclin D1 gene and protein and the expression of MMP2 protein were all increased.Conclusion UCA1 is up-regulation in breast cancer cells and may promote breast cancer proliferation,migration,and invasion by regulating cyclin D1 and MMP2.
作者
杨姝娅
刘文娟
骆耐香
韦冉
王鑫峰
刘文慧
YANG Shuya;LIU Wenjuan;LUO Naixiang;WEI Ran;WANG Xinfeng;LIU Wenhui(Department of Immunology,School of Basic Medicine,Guilin Medical University,Guilin 541001,Guangxi,China;Department of Pathology,Chengwu People’s Hospital,Heze 274200,Shandong,China;Department of Clinical Laboratory,the Second Affiliated Hospital of Guilin Medical University,Guilin 541001,Guangxi,China;Department of Human Anatomy,School of Basic Medicine,Guilin Medical University,Guilin 541001,Guangxi,China;Institute of Clinical Pharmacology,Xiangya Hospital Central South University,Changsha 410008,Hu’nan,China)
出处
《癌症进展》
2023年第9期955-960,共6页
Oncology Progress
基金
广西肿瘤免疫与微环境调控重点实验室开放课题(2020KF005)
广西高校中青年教师科研基础能力提升项目(2019KY0527)
桂林医学院中青年教职工科研能力提升项目(2018glmc099)。
