Androgen-regulated miR-32 targets BTG2 and is overexpressed in castration-resistant prostate cancer
Androgen-regulated miR-32 targets BTG2 and is overexpressed in castration-resistant prostate cancer
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DOI：
10.1038/onc.2011.624
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发表时间：
2012-10-01
期刊：
影响因子：
8
通讯作者：
Visakorpi, T.
中科院分区：
文献类型：
--
作者：
Jalava, S. E.;Urbanucci, A.;Visakorpi, T.
雄激素受体(AR)信号通路参与了去势耐受前列腺癌(CRPC)的发生。在这里，我们识别了几个可能与CRPC的发展有关的雄激素调节的microRNAs(MiRNAs)。基因芯片分析发现，7个miR-21、miR-32、miR-99a、miR-99b、miR-148a、miR-221和miR-590-5p在CRPC中的表达与良性前列腺增生症(BPH)存在差异。转染前miR-32和前miR-148A的LNCaP细胞具有明显的生长优势。MiR-32可减少细胞凋亡，而miR-148A可促进细胞增殖。雄激素对LNCaP细胞的刺激和表达分析证实了miR-32和miR-148A对雄激素的调节作用。染色质免疫沉淀(ChIP)显示了这些miRNAs附近的AR结合部位。为了确定miRNAs的靶基因，对转染前miR-32和前miR-148A的LNCaP细胞进行了mRNA微阵列分析。Pre-miR-32和Pre-miR-148A分别可降低BTG2和PIK3IP1的表达。蛋白质印迹分析显示，细胞内蛋白质表达降低。BTG2和PIK3IP1经3‘UTR-荧光素酶检测证实为靶标。免疫组织化学染色结果显示差异有统计学意义(P<0.05)。
The androgen receptor (AR) signaling pathway is involved in the emergence of castration-resistant prostate cancer (CRPC). Here, we identified several androgen-regulated microRNAs (miRNAs) that may contribute to the development of CRPC. Seven miRNAs, miR-21, miR-32, miR-99a, miR-99b, miR-148a, miR-221 and miR-590-5p, were found to be differentially expressed in CRPC compared with benign prostate hyperplasia (BPH) according to microarray analyses. Significant growth advantage for LNCaP cells transfected with pre-miR-32 and pre-miR-148a was found. miR-32 was demonstrated to reduce apoptosis, whereas miR-148a enhanced proliferation. Androgen regulation of miR-32 and miR-148a was confirmed by androgen stimulation of the LNCaP cells followed by expression analyses. The AR-binding sites in proximity of these miRNAs were demonstrated with chromatin immunoprecipitation (ChIP). To identify target genes for the miRNAs, mRNA microarray analyses were performed with LNCaP cells transfected with pre-miR-32 and pre-miR-148a. Expression of BTG2 and PIK3IP1 was reduced in the cells transfected with pre-miR-32 and pre-miR-148a, respectively. Also, the protein expression was reduced according to western blot analysis. BTG2 and PIK3IP1 were confirmed to be targets by 3'UTR-luciferase assays. Finally, immunostainings showed a statistically significant (P