MicroRNA-185 downregulates androgen receptor expression in the LNCaP prostate carcinoma cell line
MicroRNA-185 downregulates androgen receptor expression in the LNCaP prostate carcinoma cell line
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MicroRNA-185 下调 LNCaP 前列腺癌细胞系中雄激素受体的表达
DOI：
10.3892/mmr.2015.3332
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发表时间：
2015-06-01
影响因子：
3.4
通讯作者：
Jiang, Anli
中科院分区：
文献类型：
--
作者：
Liu, Chunyan;Chen, Zhaobo;Jiang, Anli
本研究旨在探讨 microRNA (miR)-185 是否下调 LNCaP 前列腺癌细胞系中雄激素受体的表达。培养人前列腺癌 (PCa) LNCaP 细胞并用合成的 has-miR-185 模拟物或抑制剂转染。随后通过活力测定、核染色、逆转录定量聚合酶链反应（RT-qPCR）、双荧光素酶测定和蛋白质印迹分析来评估转染的细胞。 Western blot分析和RT-qPCR结果表明，转染miR-185模拟物显着降低了LNCaP细胞中雄激素受体（AR）蛋白的表达水平，而转染miR-185抑制剂则增加了LNCaP细胞中AR的蛋白表达水平。荧光素酶报告基因检测结果表明，AR 3非翻译区中的预测靶位点是miR-185的特异性功能结合位点，并且AR是miR-185的直接靶点。此外，miR-185对AR的下调损害了AR与雄激素反应元件之间的相互作用，并下调了AR靶基因前列腺特异性抗原的表达。数据还表明，miR-185介导的AR下调可抑制LNCaP细胞的增殖并诱导其凋亡。因此，本研究的结果表明，miR-185可能是AR介导的信号传导的潜在负调节剂，并且可能在前列腺癌细胞中充当肿瘤抑制因子。
The present study aimed to investigate whether microRNA (miR)-185 downregulated androgen receptor expression in the LNCaP prostate carcinoma cell line. Human prostate cancer (PCa) LNCaP cells were cultured and transfected with synthetic has-miR-185 mimic or inhibitor. The transfected cells were subsequently evaluated with a viability assay, nuclear staining, reverse transcription quantitative polymerase chain reaction (RT-qPCR), dual luciferase assay and western blot analysis. The results of the western blot analysis and RT-qPCR indicated that transfection with an miR-185 mimic markedly reduced the androgen receptor (AR) protein expression levels in LNCaP cells, whereas transfection with an miR-185 inhibitor increased the protein expression of AR in the LNCaP cells. The results of the luciferase reporter assay demonstrated that the predicted target site in the AR 3 untranslated regions was a specific functional binding site for miR-185, and that AR was a direct target of miR-185. In addition, downregulation of AR by miR-185 impaired the interaction between AR and androgen response element, and downregulated the expression of the AR target gene prostate specific antigen. Data also suggested that the downregulation of AR mediated by miR-185, inhibited the proliferation and induced the apoptosis of the LNCaP cells. Therefore, the results of the present study suggested that miR-185 may be a potential negative modulator of AR-mediated signaling and may act as a tumor suppressor in prostate cancer cells.