Scaravilli Mauro, Porkka Kati P, Brofeldt Anniina, Annala Matti, Tammela Teuvo L J, Jenster Guido W, Nykter Matti, Visakorpi Tapio
Prostate Cancer Research Center, Institute of Biosciences and Medical Technology-BioMediTech and Fimlab Laboratories, University of Tampere and Tampere University Hospital, Tampere, Finland.
Prostate. 2015 Jun;75(8):798-805. doi: 10.1002/pros.22961. Epub 2015 Mar 1.
Recently, there has been increasing attention on the role of microRNAs (miRNAs) in cancer development. Several expression profiling studies have provided evidence of aberrant expression of miRNAs in prostate cancer and have highlighted the potential use of specific miRNA expression signatures as prognostic or predictive markers. Here we report an expression analysis of miR-1247-5p, miR-1249, miR-1269a, miR-1271-5p, miR-1290, miR-1291, and miR-1299.
qRT-PCR was performed to validate the differential expression of miRNAs in clinical samples, and the effect of miR-1247-5p was studied in prostate cancer cell lines transiently transfected with a miR-1247-5p mimic. The expression of miR-1247-5p's putative target MYCBP2 was evaluated by qRT-PCR and Western blotting, and the interaction of the miRNA with the target gene was assessed using a luciferase assay.
We found a significant up-regulation of miR-1247-5p in castration-resistant prostate cancer (CRPC) samples compared to non-malignant prostate. The expression of miR-1247-5p was subsequently studied in prostate cancer (PC) cell lines where an up-regulation of miR-1247-5p was observed in the androgen-independent PC-3 model. Target prediction analysis for miR-1247-5p performed online revealed that MYCBP2 (myc-binding protein 2) was a high-scoring potential target. Functional studies in vitro performed using PC-3 and LNCaP models confirmed the down-regulation of MYCBP2 at the mRNA and protein levels, and a luciferase assay showed interaction between the miRNA and target gene.
miR-1247-5p is overexpressed in CRPC and targets MYCBP2.
最近，微小RNA（miRNA）在癌症发展中的作用受到越来越多的关注。多项表达谱研究已提供证据表明miRNA在前列腺癌中表达异常，并强调了特定miRNA表达特征作为预后或预测标志物的潜在用途。在此，我们报告了miR-1247-5p、miR-1249、miR-1269a、miR-1271-5p、miR-1290、miR-1291和miR-1299的表达分析。
进行qRT-PCR以验证临床样本中miRNA的差异表达，并在瞬时转染miR-1247-5p模拟物的前列腺癌细胞系中研究miR-1247-5p的作用。通过qRT-PCR和蛋白质印迹法评估miR-1247-5p的假定靶标MYCBP2的表达，并使用荧光素酶测定法评估miRNA与靶基因的相互作用。
我们发现，与非恶性前列腺相比，去势抵抗性前列腺癌（CRPC）样本中miR-1247-5p显著上调。随后在前列腺癌（PC）细胞系中研究了miR-1247-5p的表达，在雄激素非依赖性PC-3模型中观察到miR-1247-5p上调。在线进行的miR-1247-5p靶标预测分析显示，MYCBP2（myc结合蛋白2）是一个高分潜在靶标。使用PC-3和LNCaP模型进行的体外功能研究证实了MYCBP2在mRNA和蛋白质水平上的下调，荧光素酶测定显示了miRNA与靶基因之间的相互作用。
miR-1247-5p在CRPC中过表达并靶向MYCBP2。
