HOTAIR靶向miR-513c-5p对乳腺癌增殖的调控作用及机制  

Effect and mechanism of HOTAIR on the proliferation of breast cancer cells by targeting miR-513c-5p

作　　者：钱立宇[1] 张浩然[2] 姚廷敬[1] 汤杰 刘中 岳喜成[1] QIAN Liyu;ZHANG Haoran;YAO Tingjing;TANG Jie;LIU Zhong;YUE Xicheng(Department of Surgical Oncology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Department of Medical Oncology,First Affiliated Hospital of Bengbu Medical College;Department of Hepatobiliary Surgery,First Affiliated Hospital of Bengbu Medical College)

机构地区：[1]蚌埠医学院第一附属医院肿瘤外科,蚌埠233000 [2]蚌埠医学院第一附属医院肿瘤内科 [3]蚌埠医学院第一附属医院肝胆外科

出　　处：《山西医科大学学报》2022年第5期520-525,共6页Journal of Shanxi Medical University

基　　金：蚌埠医学院自然科学重点项目(2020byzd061)。

摘　　要：目的研究HOX transcript antisense RNA(HOTAIR)通过靶向miR-513c-5p调控乳腺癌增殖的作用及机制。方法选取来自蚌埠医学院第一附属医院2020年6—12月间女性患者乳腺癌和癌旁组织各40例,应用real-time PCR方法检测癌组织和癌旁组织中HOTAIR和miR-513c-5p表达并分析其表达的相关性。沉默MCF-7细胞中HOTAIR表达建立不同HOTAIR沉默水平的sh-HOTAIR-1组和sh-HOTAIR-2组,感染空载体作为对照组。应用real-time PCR方法检测MCF-7细胞中HOTAIR和miR-513c-5p表达并分析其表达的相关性,CCK-8法分析sh-HOTAIR-1组和sh-HOTAIR-2组细胞24,48,72 h体外增殖的变化,应用流式细胞方法分析细胞周期的变化,通过蛋白印迹法(Western blot)分析细胞周期蛋白依赖性激酶(cyclin-dependent kinase,CDK)4/6蛋白表达。将miR-513c-5p抑制物转染至沉默sh-HOTAIR-2的MCF-7细胞,观察miR-513c-5p抑制物对HOTAIR沉默后MCF-7细胞增殖、细胞周期及相关蛋白表达调控的影响。结果与癌旁组织相比,乳腺癌组织HOTAIR表达显著上调,miR-513c-5p表达显著下调,二者成负相关(r=-0.5498,P<0.05)。与对照组比较,sh-HOTAIR-1组和sh-HOTAIR-2组MCF-7细胞中miR-513c-5p表达显著上调,MCF-7细胞体外增殖水平均显著下调,细胞周期G2/M期显著上调,CDK4/6蛋白和mRNA表达显著下调(均P<0.05)。与sh-HOTAIR-2组相比,sh-HOTAIR-2+miR-513c-5p抑制组中MCF-7细胞体外增殖显著上调,细胞周期G2/M期显著下调,CDK4/6蛋白和mRNA表达显著上调(均P<0.05)。结论HOTAIR在乳腺癌中高表达,可促进肿瘤细胞体外增殖,可能与抑制miR-513c-5p表达有关。Objective To investigate the effect and mechanism of HOX transcript antisense RNA(HOTAIR)on the proliferation of breast cancer cells by targeting miR-513 c-5 p.Methods A total of 40 breast cancer tissues and 40 paraneoplastic tissues were obtained from the First Affiliated Hospital of Bengbu Medical College from June to December in 2020.The expression of HOTAIR and miR-513 c-5 p was investigated in breast cancer tissues and paraneoplastic tissues by real-time PCR,and their correlation was analyzed.HOTAIR was silenced in MCF-7 cells,and then MCF-7 cells were divided into sh-HOTAIR-1 group and sh-HOTAIR-2 group according to different HOTAIR,and MCF-7 cells were transfected with blank vector in control group.The expression of HOTAIR and miR-513 c-5 p was measured in MCF-7 cells by real time PCR assay and their correlation was analyzed.The cell proliferation in 24,48,72 h,the cell cycle,and the expression of cyclin-dependent kinase(CDK)4/6 were measured after silencing HOTAIR by CCK-8 assay,flow cytometry and Western blot assay,respectively.The miR-513 c-5 p inhibitor was transfected into MCF-7 cells in sh-HOTAIR-2 group,and the effects of miR-513 c-5 p on the proliferation,the cell cycle,and the related protein expression were observed in MCF-7 cells.Results Compared with paraneoplastic tissues,the expression of HOTAIR was up-regulated in breast cancer tissue,and the expression of miR-513 c-5 p was down-regulated,and the expression of HOTAIR was negatively correlated with miR-513 c-5 p expression(r=-0.5498,P<0.05).Compared with control group,the expression of miR-513 c-5 p was an up-regulated in sh-HOTAIR-1 and sh-HOTAIR-2 group,the proliferation of MCF-7 cells was down-regulated,the G2/M cells were up-regulated,and the expression of CDK 4/6 was down-regulated(all P<0.05).Compared with sh-HOTAIR-2 group,the proliferation was significantly up-regulated,the G2/M cells were significantly down-regulated,and the expression of CDK4/6 protein and mRNA was significantly up-regulated in sh-HOTAIR-2+miR-513 c-5 p inhibitor gr

关 键 词：HOTAIR 乳腺癌 细胞增殖 

分 类 号：R737.9[医药卫生—肿瘤]