摘要
目的观察微小RNA(miRNA,miR)-26a对三阴性乳腺癌细胞增殖的影响,并探讨其机制。方法通过噻唑蓝(MTT)实验及集落形成实验检测miR-26a对细胞增殖能力的影响,应用流式细胞技术检测miR-26a对细胞周期的影响。利用数据库miRTarBase预测miR-26a靶基因,在数百种靶基因中,筛选出与细胞周期及肿瘤转移密切相关的靶基因c-Myc作为研究对象。通过Western blot实验检测miR-26a过表达后细胞中c-Myc及其下游靶基因细胞周期蛋白D2(Cyclin D2)、细胞周期蛋白E1(Cyclin E1)蛋白水平的表达。结果MTT实验显示,miR-26a模拟物(mimic)转染组细胞的增殖活力显著低于miR-mimic阴性对照(NC)转染组的细胞。集落形成实验显示,miR-26a mimic转染组细胞的克隆增殖能力显著低于miR-mimic NC转染组的细胞[(55.67±5.86)个比(144.33±5.03)个;(62.00±3.00)个比(107.67±2.52)个,P<0.01]。流式细胞技术显示,miR-26a mimic转染组细胞处于G1期的细胞比例明显高于miR-mimic NC转染组的细胞[(59.39±1.04)%比(46.91±1.04)%;(76.91±1.43)%比(63.21±1.61)%,P<0.01]。Western blot实验显示,细胞中miR-26a表达上调后,c-Myc及其下游靶基因Cyclin D2、Cyclin E1的蛋白表达水平显著降低。结论 miR-26a可抑制三阴性乳腺癌的增殖,其机制可能是miR-26a通过靶向癌基因c-Myc来发挥抑癌作用。
Objective To observe the effect and mechanism of microRNA (miRNA, miR)-26a on the proliferation of triple negative breast cancer cells.Methods The effect of miR-26a on cell proliferation was detected by methyl thiazol tetrazolium (MTT) assay and colony formation assay, the effect of miR-26a on cell cycle was detected by flow cytometry. Use miRTarBase to predict target genes of miR-26a. In hundreds of target genes, we screened oncogene c-Myc as the research object. Use Western blotting test to detect the expression of c-Myc and its downstream target genes Cyclin D2 and Cyclin E1 in triple negative breast cancer cell lines after miR-26a overexpression.Results MTT assay revealed that the proliferation activity of miR-26a mimic transfection group was significantly lower than that of miR-mimic NC transfection group. Colony formation assay revealed that the cell clone ability of miR-26a mimic transfection group was significantly lower than that of miR-mimic NC transfection group [(55.67±5.86) cells vs. (144.33±5.03) cells;(62.00±3.00) cells vs. (107.67±2.52) cells, P<0.01]. Flow cytometry showed that the proportion of cells in G1 phase in the miRNA-26a mimic transfection group was significantly higher than that in the miRNA-mimic NC transfection group [(59.39±1.04)% vs. (46.91±1.04)%;(76.91±1.43)% vs. (63.21±1.61)%, P<0.01]. Western blotting test revealed that the expression of c-Myc and its downstream target genes Cyclin D2 and Cyclin E1 were significantly decreased.Conclusion MiR-26a may inhibit the proliferation of triple negative breast cancer by targeting the oncogene c-Myc.
作者
解荣
李继勇
喻远航
张悦
廖晗
张波
Xie Rong;Li Jiyong;Yu Yuanhang;Zhang Yue;Liao Han;Zhang Bo(Department of Breast and Thyroid Surgery,Union Hospital,Tong]i Medical College,Huazhong University of Science and Technology,Wuhan 430022,China;Department of Breast and Thyroid Surgery,Huangpi People's Hospital,Wuhan (the Affiliated Huangpi People's Hospital of Jianghan University),Wuhan 430300.China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第1期63-65,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81472487、81773197).
