miR-451a Inhibited Cell Proliferation and Enhanced Tamoxifen Sensitive in Breast Cancer via Macrophage Migration Inhibitory Factor.
miR-451a Inhibited Cell Proliferation and Enhanced Tamoxifen Sensitive in Breast Cancer via Macrophage Migration Inhibitory Factor.
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DOI：
10.1155/2015/207684
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发表时间：
2015
影响因子：
--
通讯作者：
Li H
中科院分区：
文献类型：
--
作者：
Liu Z;Miao T;Feng T;Jiang Z;Li M;Zhou L;Li H
本研究旨在探讨microRNA-451 a（miR-451 a）对乳腺癌细胞增殖和对他莫昔芬敏感性的调节作用。在细胞培养实验中，构建了pHBLV-miR-451 a和pHBLV-miR-451 a海绵慢病毒载体，并用于转染MCF-7和LCC 2细胞。通过荧光观察检测转染效率，建立稳定过表达或下调表达miR-451 a的细胞系。采用实时荧光定量逆转录聚合酶链反应（real-time reverse transcriptase polymerase chain reaction，RT-PCR）和/或western blot检测miR-451 a和靶基因巨噬细胞移动抑制因子（macrophage migration inhibitor factor，MIF）的表达。此外，还进行了MTT测定、集落形成和Transwell侵袭测定。结果表明，重组慢病毒载体构建正确，病毒滴度为1 × 108 CFU/mL。获得稳定转染的细胞。miR-451 a过表达下调MIF在mRNA和蛋白水平的表达，并抑制乳腺癌细胞的增殖、集落形成和侵袭。下调miR-451 a可上调MIF表达，并增加乳腺癌细胞的生长、侵袭和他莫昔芬敏感性。综上所述，miR-451 a/MIF通路可能在乳腺癌细胞的生物学特性中发挥重要作用，并可能成为乳腺癌潜在的治疗靶点。
This study aims to investigate the regulative effects of microRNA-451a (miR-451a) on cell proliferation and sensitivity to tamoxifen in breast cancer cells. In cell culture experiments, the lentiviral vectors of pHBLV-miR-451a and pHBLV-miR-451a sponge were constructed and used to transfect MCF-7 and LCC2 cells. The transfection efficiency was tested by fluorescent observation, and cell lines with stable over- or downregulated expression of miR-451a were established. The expression of miR-451a and the target gene macrophage migration inhibitory factor (MIF) were detected by real-time reverse transcriptase polymerase chain reaction and/or western blot. Moreover, MTT assay, colony formation, and Transwell invasion assays were also performed. Data showed that the recombinant lentiviral vectors were constructed correctly, and the virus titer was 1 × 108 CFU/mL. The stable transfected cells were obtained. Overexpression of miR-451a downregulated MIF expression in mRNA and protein levels and inhibited cell proliferation, colony formation, and invasion of breast cancer cells. Downregulation of miR-451a upregulated MIF expression and increased breast cancer cell growth, invasion, and tamoxifen sensitivity. In summary, the miR-451a/MIF pathway may play important roles in the biological properties of breast cancer cells and may be a potential therapeutic target for breast cancer.
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