Voth Harald, Oberthuer André, Simon Thorsten, Kahlert Yvonne, Berthold Frank, Fischer Matthias
Department of Pediatric Oncology and Hematology, University Children's Hospital of Cologne, Kerpener Strasse 62, 50924 Cologne, Germany.
Mol Cancer Res. 2007 Dec;5(12):1276-84. doi: 10.1158/1541-7786.MCR-06-0258.
Neuroblastoma at stage IVS, defined by dissemination to specific tissues and age <1 year at diagnosis, regularly follows spontaneous regression without cytotoxic treatment. To uncover the molecular characteristics of this subtype, Serial Analysis of Gene Expression (SAGE) profiles from stage IVS and fatal stage IV tumors were compared. A SAGE tag (GCAACTTAAC) was detected that was overrepresented in stage IVS disease and that had no reliable match in current National Center for Biotechnology Information databases of SAGE profiles, thus pointing to a novel gene. The corresponding gene, which maps to chromosome 4q33-34, was identified using a modified 3'- and 5'-rapid amplification of cDNA ends PCR and was designated as DEIN (differentially expressed in neuroblastoma). The gene comprises five transcript variants and its sequence overlaps with expressed sequences of the yet uncharacterized UniGene cluster Hs.61435. DEIN exhibits nucleotide sequence conservation over a broad range of species with an overall homology of 65% between human and mouse. As none of the predicted amino acid sequences is homologous to known proteins, it remains to be determined whether DEIN represents a coding or noncoding RNA. Northern blot analysis and semiquantitative reverse transcription-PCR showed high DEIN expression in neuroblastoma, whereas expression was absent or weak in most normal adult tissues. Analysis of 121 primary neuroblastomas by real-time reverse transcription-PCR revealed a strong association with age at diagnosis <1 year and particularly with stage IVS disease (both P < 0.001). The characteristic expression pattern of DEIN suggests a specific role of this gene in the unique biology of stage IVS tumors and may help to molecularly define this special subtype of neuroblastoma.
IVS期神经母细胞瘤定义为肿瘤扩散至特定组织且诊断时年龄小于1岁，通常在无细胞毒性治疗的情况下会自然消退。为了揭示该亚型的分子特征，对IVS期和致命IV期肿瘤的基因表达系列分析（SAGE）图谱进行了比较。检测到一个SAGE标签（GCAACTTAAC），其在IVS期疾病中过度表达，且在当前美国国立生物技术信息中心的SAGE图谱数据库中没有可靠的匹配项，因此指向一个新基因。使用改良的3'和5' cDNA末端快速扩增PCR鉴定了位于染色体4q33 - 34上的相应基因，并将其命名为DEIN（在神经母细胞瘤中差异表达）。该基因包含五个转录变体，其序列与尚未表征的UniGene簇Hs.61435的表达序列重叠。DEIN在广泛的物种中表现出核苷酸序列保守性，人与小鼠之间的总体同源性为65%。由于预测的氨基酸序列均与已知蛋白质无同源性，因此DEIN是编码RNA还是非编码RNA仍有待确定。Northern印迹分析和半定量逆转录PCR显示DEIN在神经母细胞瘤中高表达，而在大多数正常成人组织中无表达或表达较弱。通过实时逆转录PCR对121例原发性神经母细胞瘤进行分析，发现其与诊断时年龄小于1岁尤其是与IVS期疾病密切相关（P均<0.001）。DEIN的特征性表达模式表明该基因在IVS期肿瘤的独特生物学特性中具有特定作用，可能有助于从分子水平定义神经母细胞瘤的这一特殊亚型。
