Research Division, DCD, Inc., Pohang Technopark, Pohang, Kyungbuk, 790-834, Korea.
J Proteome Res. 2010 Jul 2;9(7):3710-9. doi: 10.1021/pr100236r.
The timely diagnosis and therapeutic monitoring of human renal cell carcinoma (RCC) is limited by the lack of specific biomarkers. To identify candidate RCC biomarkers, we used 2-DE gel electrophoresis with mass spectrometry and 2-DE spot intensity-based ROC analysis to analyze 18 sets of paired normal and RCC tumor tissue including conventional, papillary, and chromophobe subtypes. Validation was performed with RCC patient plasma samples and confirmed by clustergram, shRNA, and immunohistochemistry assays. Cardinal candidates were evaluated by ELISA. The leading candidate biomarker that was upregulated in RCC samples according to the clustergram and validation analysis was nicotinamide N-methyltransferase (NNMT) (13/15, P < 0.0001). Other upregulated candidate biomarkers that were identified by this method include ferritin, hNSE, NM23, secretagogin, and L-plastin. The upregulation of NNMT in RCC was confirmed by immunoblotting and immunohistochemistry. Analysis of fractionated membrane-associated proteins identified CAP-G, mitofillin, tubulin alpha, RBBP7, and HSP27. Of these, RBBP7 and HSP27 were highly expressed in the chromophobe subtype of RCC (3/3) but were absent from conventional RCC (0/3). The triple combination of the NNMT, FTL, and hNSE biomarkers had the highest predictive capacity of 0.993, while NNMT was the single, most powerful candidate diagnostic biomarker for all types of RCC.
人类肾细胞癌 (RCC) 的及时诊断和治疗监测受到缺乏特异性生物标志物的限制。为了鉴定候选 RCC 生物标志物，我们使用 2-DE 凝胶电泳联合质谱和基于 2-DE 斑点强度的 ROC 分析，分析了包括常规型、乳头状型和嫌色细胞型在内的 18 对配对的正常和 RCC 肿瘤组织。使用 RCC 患者的血浆样本进行验证，并通过聚类分析、shRNA 和免疫组织化学检测进行确认。通过 ELISA 评估主要候选标志物。根据聚类分析和验证分析，上调的 RCC 样本中的主要候选生物标志物是烟酰胺 N-甲基转移酶 (NNMT) (13/15, P < 0.0001)。通过这种方法鉴定的其他上调候选生物标志物包括铁蛋白、hNSE、NM23、分泌素和 L-肌动蛋白。免疫印迹和免疫组织化学证实了 NNMT 在 RCC 中的上调。膜相关蛋白的分级分析鉴定了 CAP-G、肌联蛋白、微管蛋白α、RBBP7 和 HSP27。其中，RBBP7 和 HSP27 在 RCC 的嫌色细胞型中高度表达 (3/3)，但在常规 RCC 中不存在 (0/3)。NNMT、FTL 和 hNSE 生物标志物的三联组合具有最高的预测能力 0.993，而 NNMT 是所有类型 RCC 的单一、最有力的候选诊断生物标志物。
