视网膜母细胞瘤oncomiRs的鉴定及其功能研究

项目名称： 视网膜母细胞瘤oncomiRs的鉴定及其功能研究

项目编号： No.30872843

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 阳菊华

作者单位： 福建医科大学

项目金额： 32万元

中文摘要： 本项目采用miR芯片技术构建了视网膜母细胞瘤和正常视网膜的miR表达谱，结果显示肿瘤中miRs表达总体趋势下调，其中异常表达的miRs常以基因簇形式呈现，且位于比较靠近RB 基因组不稳定区域，如miR-17-92位于靠近13q32-q34 的MRG，miR-517/518位于靠近19q13.2-13.3 的 MDR。采用Northern blot 和组织芯片-原位杂交等方法验证了一些RB 肿瘤相关miRs 及其视网膜组织定位，如miR-320在RB组织明显高表达，在正常视网膜组织中其主要分布神经节细胞层和内核层。qRT-PCR分析结果表明在RB组织和RB细胞系中MCM7及其内含的miR-106b-25 基因簇都高表达。构建和包装siRNA-MCM7慢病毒，感染Y79细胞可干扰MCM7的表达，明显抑制细胞的生长，可能与细胞凋亡有关。结论：RB基因组不稳定性与其miR异常表达相关，miR在RB肿瘤的发生/发展中起着协同者的作用或起着"加速器"的作用，利用miR药物靶分子治疗肿瘤很难取得理想的效果。

中文关键词： 视网膜母细胞瘤；Y79细胞；microRNA；oncomiRs；miR靶基因

英文摘要： In this project，using microRNA microarray technique, the microRNA profiles were constructed in human retinoblastoma and normal retina tissue. The results showed that the overall trend of miRs expression is down-regulated in RB tumor, and these abnormal expressed miRs，usually with gene clusters, were often located in RB genome instabiity region.For example, the miR-17~92 cluster was closed to minimal common region of gain on chromosom 13 q32-q34, and the miR-517/518 cluster was to minimal deleted regions on chromosom 19q13.2-13.3. some difference miRNAs were verified using northern blot analysis and the tissue microarray immunohistochemical analysis. For example, miR-320 was overexpresed in RB tissues, and it was mainly expressed in the cytoplasm of the ganglion cell layer and the inner nuclear layer (INL) of normal retinal tisuuse. The miR-106b~25 cluster and its host gene MCM7 were overexpressed in retinoblatoma tissuse and cell lines(Y79 and Hxo-Rb44). The siRNA-MCM7 lentivirus was constructed and pakaged, and then was infected in Y79 cell. The MCM7 expression was down-regulated by siRNA and the growth of infected cells was inhibited, which may be concerned with cell apoptosis. Conclusion: The abnormal expression of miRs was involved in genome instabiity region in retinoblastoma, and these miRs could cooperate with other genes in the process of RB development. In addtion, it may be difficult to achieve the ideal effect using miR as drug target molecule on RB therapy.

英文关键词： retinoblastoma；Y79 cell line；microRNA；oncomiRs；miR-target