Hsu Wen-Ming, Lee Hsinyu, Juan Hsueh-Fen, Shih Yu-Yin, Wang Bo-Jeng, Pan Chien-Yuan, Jeng Yung-Ming, Chang Hsiu-Hao, Lu Meng-Yao, Lin Kai-Hsin, Lai Hong-Shiee, Chen Wei-Jao, Tsay Yeou-Guang, Liao Yung-Feng, Hsieh Fon-Jou
Department of Surgery, National Taiwan University Hospital and National Taiwan University College of Medicine, 7 Chung-Shan South Road, Taipei, Taiwan.
Clin Cancer Res. 2008 Oct 1;14(19):6237-45. doi: 10.1158/1078-0432.CCR-07-4181.
Neuroblastoma (NB) is a heterogeneous neoplasm. Detailed biological discrimination is critical for the effective treatment of this disease. Because the tumor behavior of NB is closely associated with the histologic state of differentiation, we thus aimed to identify novel differentiation-associated markers of NB with prognostic implication.
A human NB cell line SH-SY5Y was used as a model system to explore potential biomarkers for the differentiation of NB by proteomic analyses. Seventy-two NB tumor tissues were subsequently investigated by immunohistochemistry to validate the correlations between the expression of a novel prognostic marker, various clinicopathologic and biological factors, and patient survival.
Using two-dimensional differential gel electrophoresis, we found a total of 24 spots of proteins in SH-SY5Y cells whose expression was enhanced following differentiation. Glucose-regulated protein 75 (GRP75) was unambiguously identified as one of the five proteins that were dramatically up-regulated following differentiation. Immunohistochemical analyses of 72 NB tumor tissues further revealed that positive GRP75 immunostaining is strongly correlated with differentiated histologies (P < 0.001), mass-screened tumors (P = 0.016), and early clinical stages (P < 0.001) but inversely correlated with MYCN amplification (P = 0.010). Univariate and multivariate survival analyses showed that GRP75 expression is an independent favorable prognostic factor.
The present findings clearly showed that our proteomics-based novel experimental paradigm could be a powerful tool to uncover novel biomarkers associated with the differentiation of NB. Our data also substantiate an essential role of GRP75 in the differentiation of NB.
神经母细胞瘤（NB）是一种异质性肿瘤。详细的生物学鉴别对于有效治疗该疾病至关重要。由于NB的肿瘤行为与组织学分化状态密切相关，因此我们旨在鉴定具有预后意义的新型NB分化相关标志物。
使用人NB细胞系SH-SY5Y作为模型系统，通过蛋白质组学分析探索NB分化的潜在生物标志物。随后通过免疫组织化学对72例NB肿瘤组织进行研究，以验证新型预后标志物的表达、各种临床病理和生物学因素与患者生存之间的相关性。
使用二维差异凝胶电泳，我们在SH-SY5Y细胞中总共发现了24个蛋白质斑点，其表达在分化后增强。葡萄糖调节蛋白75（GRP75）被明确鉴定为分化后显著上调的五种蛋白质之一。对72例NB肿瘤组织的免疫组织化学分析进一步显示，GRP75免疫染色阳性与分化组织学（P < 0.001）、大规模筛查肿瘤（P = 0.016）和早期临床阶段（P < 0.001）密切相关，但与MYCN扩增呈负相关（P = 0.010）。单因素和多因素生存分析表明，GRP75表达是一个独立的有利预后因素。
本研究结果清楚地表明，我们基于蛋白质组学的新型实验范式可能是揭示与NB分化相关的新型生物标志物的有力工具。我们的数据也证实了GRP75在NB分化中的重要作用。
