Chen Po, Xu Wentao, Luo Yi, Zhang Yi, He Yi, Yang Shuo, Yuan Zhijun
Department of Medical Oncology, Hunan Cancer Hospital, Changsha.
Clinical Medical College of An Hui Medical University, Hefei.
Onco Targets Ther. 2017 Mar 6;10:1423-1431. doi: 10.2147/OTT.S118366. eCollection 2017.
Breast cancer affects millions of people with a high mortality rate throughout the world; microRNA 543 (miR-543) has been reported to suppress progression in some kinds of cancers, but has not been reported in breast cancer. Thus, the purpose of this study is to investigate the function of miR-543 in breast cancer cells.
Two cell lines, MCF-7 and MDA-MB-231, were selected to be the research objects; the miR-543 overexpression and knockdown models were established in the study by transforming miR-543 mimics and miR-543 inhibitor. Real-time polymerase chain reaction, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Western blot, clone formation and cell flow cytometer assay were used to test the miR-543's function. Dual-luciferase assay was used for the detection of miR-543 and ERK2 targeting relationship.
The results showed that the cell proliferation and cell cycle were inhibited, and the capability of cell apoptosis was upregulated when miR-543 was overexpressed; we found that there was a target relationship between ERK2 and miR-543. Furthermore, downstream factors of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase-2 (ERK2) pathway, including RSK2 and MSK1, were decreased in miR-543 overexpression model.
This study provides series evidences to support that breast cancer progression was inhibited by miR-543 via direct targeting of ERK2 in MAPK/ERK signal pathway, which may provide a molecular basis for better treatment for patients.
乳腺癌在全球影响着数百万人,死亡率很高;据报道,微小RNA 543(miR-543)可抑制某些类型癌症的进展,但在乳腺癌中尚未见报道。因此,本研究旨在探讨miR-543在乳腺癌细胞中的功能。
选择MCF-7和MDA-MB-231两种细胞系作为研究对象;通过转染miR-543模拟物和miR-543抑制剂,在本研究中建立了miR-543过表达和敲低模型。采用实时聚合酶链反应、3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐、蛋白质免疫印迹法、克隆形成和细胞流式细胞仪检测来测试miR-543的功能。采用双荧光素酶报告基因检测法检测miR-543与ERK2的靶向关系。
结果显示,miR-543过表达时,细胞增殖和细胞周期受到抑制,细胞凋亡能力上调;我们发现ERK2与miR-543之间存在靶向关系。此外,在miR-543过表达模型中,丝裂原活化蛋白激酶(MAPK)/细胞外信号调节激酶-2(ERK2)信号通路的下游因子,包括RSK2和MSK1,均减少。
本研究提供了一系列证据支持miR-543通过直接靶向MAPK/ERK信号通路中的ERK2来抑制乳腺癌进展,这可能为患者的更好治疗提供分子基础。