Ayers Duncan, Mestdagh Pieter, Van Maerken Tom, Vandesompele Jo
Centre for Molecular Medicine and Biobanking, University of Malta, Msida, Malta ; Manchester Institute of Biotechnology, Faculty of Medical and Human Sciences, The University of Manchester, United Kingdom.
Center for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
Comput Struct Biotechnol J. 2015 Apr 29;13:307-19. doi: 10.1016/j.csbj.2015.04.003. eCollection 2015.
The emergence of the role of microRNAs (miRNAs) in exacerbating drug resistance of tumours is recently being highlighted as a crucial research field for future clinical management of drug resistant tumours. The purpose of this study was to identify dys-regulations in expression of individual and/or networks of miRNAs that may have direct effect on neuroblastoma (NB) drug resistance.
Individual subcultures of chemosensitive SH-SY5Y and UKF-NB-3 cells were rendered chemoresistant to doxorubicin (SH-SY5Y, UKF-NB-3) or etoposide (SH-SY5Y). In each validated chemoresistance model, the parental and subcultured cell lines were analysed for miRNA expression profiling, using a high-throughput quantitative polymerase chain reaction (RT-qPCR) miRNA profiling platform for a total of 668 miRNAs.
A unique expression signature of miRNAs was found to be differentially expressed (higher than 2-fold change) within all three NB chemoresistance models. Four miRNAs were upregulated in the subcultured chemoresistant cell line. Three miRNAs were found to be downregulated in the chemoresistant cell lines for all models.
Based on the initial miRNA findings, this study elucidates the dys-regulation of four miRNAs in three separate NB chemoresistant cell line models, spanning two cell lines (SH-SY5Y and UKF-NB-3) and two chemotherapeutic agents (doxorubicin and etoposide). These miRNAs may thus be possibly linked to chemoresistance induction in NB. Such miRNAs are good candidates to be novel drug targets for future miRNA based therapies against aggressive tumours that are not responding to conventional chemotherapy.
微小RNA（miRNA）在加剧肿瘤耐药性方面的作用正日益成为肿瘤耐药性未来临床管理的关键研究领域。本研究的目的是确定可能对神经母细胞瘤（NB）耐药性有直接影响的单个和/或miRNA网络表达失调情况。
对化疗敏感的SH-SY5Y和UKF-NB-3细胞的单个亚培养物进行处理，使其对多柔比星（SH-SY5Y、UKF-NB-3）或依托泊苷（SH-SY5Y）产生耐药性。在每个经过验证的耐药模型中，使用高通量定量聚合酶链反应（RT-qPCR）miRNA分析平台对总共668种miRNA分析亲代和亚培养细胞系的miRNA表达谱。
在所有三种NB耐药模型中发现了独特的miRNA表达特征，其差异表达（变化超过2倍）。在亚培养的耐药细胞系中有四种miRNA上调。在所有模型的耐药细胞系中发现三种miRNA下调。
基于最初的miRNA研究结果，本研究阐明了三种单独的NB耐药细胞系模型中四种miRNA的表达失调，这些模型涉及两种细胞系（SH-SY5Y和UKF-NB-3）和两种化疗药物（多柔比星和依托泊苷）。因此，这些miRNA可能与NB中的耐药诱导有关。此类miRNA是未来基于miRNA的疗法针对对传统化疗无反应的侵袭性肿瘤的新型药物靶点的良好候选者。
