Guo Fei, Zhang Junjie, Wang Lei, Zhao Wei, Yu Jiekai, Zheng Shu, Wang Jiaxiang
Department of Pediatric Surgery, The First Affiliated Hospital, Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Institute of Cancer, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310000, P.R. China.
Oncol Lett. 2017 Jul;14(1):687-694. doi: 10.3892/ol.2017.6261. Epub 2017 May 26.
The present study aimed to identify differentially expressed inflammatory factors observed in Wilms tumors (WT), and to investigate the association of these factors with clinical stage, pathological type, lymph node metastasis and vascular involvement of WT. Surface-enhanced laser desorption/ionization-time of flight mass spectrometry was performed to screen differentially expressed proteins among WT and normal tissue pairs. Upregulated proteins in WT were separated and purified by solid phase extraction and Tricine SDS-PAGE, respectively. Following in-gel digestion, the peptide mixture was subjected to liquid chromatography mass spectrometry to identify proteins on the basis of their amino acid sequences. Immunohistochemistry was used to confirm the expression of differentially expressed inflammatory proteins. Of the proteins that were upregulated in WT, two proteins with mass/charge (m/z) ratio of 12,138 and 13,462 were identified as macrophage migration inhibitory factor (MIF) and C-X-C motif ligand 7 (CXCL7) chemokine, respectively. The expression of these two proteins was increased in WT compared with adjacent normal tissues and normal renal tissues, and increased with increasing clinical stage. In addition, their expression was significantly increased in patients with unfavorable pathological type, lymph node metastasis and vascular involvement compared with the groups with favorable type, and without lymph node metastasis or vascular involvement (P<0.05). Increased pro-inflammatory MIF and CXCL7 expression in WT is closely associated with the clinical stage, pathological type, lymph node metastasis and vascular involvement, and may represent biomarkers for the clinical diagnosis of WT.
本研究旨在鉴定在肾母细胞瘤（WT）中观察到的差异表达炎症因子，并探讨这些因子与WT临床分期、病理类型、淋巴结转移和血管受累情况的关联。采用表面增强激光解吸/电离飞行时间质谱法筛选WT与正常组织对之间差异表达的蛋白质。WT中上调的蛋白质分别通过固相萃取和Tricine SDS-PAGE进行分离和纯化。凝胶内消化后，将肽混合物进行液相色谱质谱分析，根据氨基酸序列鉴定蛋白质。采用免疫组织化学法确认差异表达炎症蛋白的表达。在WT中上调的蛋白质中，质荷比（m/z）为12138和13462的两种蛋白质分别被鉴定为巨噬细胞迁移抑制因子（MIF）和C-X-C基序配体7（CXCL7）趋化因子。与相邻正常组织和正常肾组织相比，这两种蛋白质在WT中的表达增加，且随临床分期的增加而增加。此外，与病理类型良好、无淋巴结转移或血管受累的组相比，病理类型不良、有淋巴结转移和血管受累的患者中它们的表达显著增加（P<0.05）。WT中促炎MIF和CXCL7表达的增加与临床分期、病理类型、淋巴结转移和血管受累密切相关，可能代表WT临床诊断的生物标志物。
