Department of Molecular and Cellular Therapeutics, Cancer Genetics Research Group, Royal College of Surgeons in Ireland, York House, York Street, Dublin 2, Ireland.
Br J Cancer. 2012 Sep 4;107(6):967-76. doi: 10.1038/bjc.2012.356. Epub 2012 Aug 14.
Neuroblastoma remains a major cause of cancer-linked mortality in children. miR-204 has been used in microRNA expression signatures predictive of neuroblastoma patient survival. The aim of this study was to explore the independent association of miR-204 with survival in a neuroblastoma cohort, and to investigate the phenotypic effects mediated by miR-204 expression in neuroblastoma.
Neuroblastoma cell lines were transiently transfected with miR-204 mimics and assessed for cell viability using MTS assays. Apoptosis levels in cell lines were evaluated by FACS analysis of Annexin V-/propidium iodide-stained cells transfected with miR-204 mimics and treated with chemotherapy drug or vehicle control. Potential targets of miR-204 were validated using luciferase reporter assays.
miR-204 expression in primary neuroblastoma tumours was predictive of patient event-free and overall survival, independent of established known risk factors. Ectopic miR-204 expression significantly increased sensitivity to cisplatin and etoposide in vitro. miR-204 direct targeting of the 3' UTR of BCL2 and NTRK2 (TrkB) was confirmed.
miR-204 is a novel predictor of outcome in neuroblastoma, functioning, at least in part, through increasing sensitivity to cisplatin by direct targeting and downregulation of anti-apoptotic BCL2. miR-204 also targets full-length NTRK2, a potent oncogene involved with chemotherapy drug resistance in neuroblastoma.
神经母细胞瘤仍然是导致儿童癌症相关死亡的主要原因。miR-204 已被用于预测神经母细胞瘤患者生存的 microRNA 表达特征。本研究旨在探讨 miR-204 与神经母细胞瘤患者生存的独立相关性，并研究 miR-204 表达对神经母细胞瘤的表型影响。
瞬时转染 miR-204 模拟物，使用 MTS 测定法评估神经母细胞瘤细胞系的细胞活力。通过转染 miR-204 模拟物并用化疗药物或载体对照处理的 Annexin V-/碘化丙啶染色细胞的 FACS 分析评估细胞系中的凋亡水平。使用荧光素酶报告基因测定法验证 miR-204 的潜在靶标。
原发性神经母细胞瘤肿瘤中的 miR-204 表达可预测患者无事件和总生存，独立于已建立的已知危险因素。体外异位 miR-204 表达显着增加顺铂和依托泊苷的敏感性。证实了 miR-204 对 BCL2 和 NTRK2（TrkB）3'UTR 的直接靶向作用。
miR-204 是神经母细胞瘤预后的新预测因子，至少部分通过直接靶向和下调抗凋亡 BCL2 来增加顺铂的敏感性起作用。miR-204 还靶向全长 NTRK2，这是一种与神经母细胞瘤中化疗药物耐药性相关的强效致癌基因。
