miRNAs的表达在神经母细胞瘤转移过程中的作用及意义

Identification of miRNAs based on microarrays in human metastatic and primary neuroblastoma

目的检测荷瘤鼠的神经母细胞瘤（neuroblastoma，NB）原发瘤与转移瘤中微小RNA（miRNA）的表达差异情况，探讨异常表达的miRNA参与神经母细胞瘤发生侵袭和转移的可能分子机制。方法建立神经母细胞瘤原发瘤和转移瘤的裸鼠模型，获取原发瘤和转移瘤的组织，分别提取总RNA和小分子RNA，在miRNA微阵列芯片中杂交检测，通过芯片扫描和数据分析并通过实时荧光定量Rrr_PCR验证，获得神经母细胞瘤原发瘤与转移瘤异常miRNAs表达谱。结果筛选出54个可能与神经母细胞瘤侵袭和转移的相关的miRNAs，表达上调35个，下调19个。其中，miR-483—3p、miR-99b、miR-145、miR602、miR-26b、miR125a-3p、let-7f在神经母细胞瘤原发瘤中与转移瘤中的表达的差异倍数分别为9．9583、3．2086、4．9442、4．4085、0．1380、0．1463和0．2864，基因芯片显著性分析指数分别为26．8754、22．8393、22．2147、8．5906、-24．5380、-10．5580和-12．6103，差异有统计学意义（P〈O．01）。结论筛选出的在神经母细胞瘤原发瘤与转移瘤中表达有差异的miRNAs可能在神经母细胞瘤的侵袭和转移过程中发挥着重要作用，为研究神经母细胞瘤的转移和治疗提供了新的靶点。

Objective To investigate the difference in the expressions of microRNAs （miRNAs） between metastatic neuroblastoma（NB） and primary NB, and to explore the molecular mechanisms of the metastasis and invasion of NB influenced by abnormal expressions of miRNAs. Methods The tis- sues of metastatic NB and primary NB were obtained from nude mice. The miRNAs were extracted and isolated. Hybridization was carried out using miRNA microarray chips, followed by the real-time quan- titative RT-PCR analysis. R^u｜ts Fifty-five miRNAs were identified in metastatic NB, 35 of which were up-regulated and 19 of which were down-regulated. The expressions of miR-483-3p, miR-99b, miR-145,miR-602, miR.-26b, miR-125a-3p and let-7f in the primary NB were significantly different from those in the metastatic NB. Conclusions The aberrant expressions of miRNAs may be involved in the pathogenesis of metastasis of NB.