荧光差异双向凝胶电泳筛选肾透明细胞癌及癌旁组织中的差异表达蛋白

目的 寻找肾癌相关肿瘤标记物,用于肾癌临床诊断、药物治疗靶点的选择及预后监测.方法 采用荧光差异双向凝胶电泳技术筛选15例肾透明细胞癌及癌旁正常肾组织差异表达蛋白质点.统计学采用t检验进行定量比较,以两组蛋白含量比值在1.5倍及以上,P<0.05,被认为是差异表达蛋白质点.用基质辅助激光解吸电离飞行时间质谱或串联质谱技术进行差异表达蛋白质点鉴定.结果 共筛选分离出27个差异表达蛋白质点,经质谱技术共成功鉴定了26种蛋白质,11种蛋白质在肾癌组织中上调,15种蛋白质下调.其功能覆盖细胞活动的多个方面,其中线粒体短/支链特异性酰基辅酶A脱氢酶、醛糖1-表异构酶、PRDX4、CAPG、β-防御素107、微纤丝相关糖蛋白4等6种蛋白是我们首次采用蛋白质组学技术成功筛选的肾癌差异表达蛋白质,并发现了1个预测蛋白(UCRIL).结论 荧光差异双向凝胶电泳在肾癌组织中获得较好的筛选效果,新筛选的差异表达蛋白有望成为肾癌分子生物学标记物,为后续研究提供了实验依据.

Quantitative and comparative proteomics analysis in clear cell renal cell carcinoma and adjacent noncancerous tissues by 2-D DIGE

Objective To identify specific protein markers for renal cell carcinoma detection and diagnosis, as well as develop new potential therapeutic targets of the disease. Methods We used two-dimensional difference in-gel electrophoresis (2-D DIGE) technique conjunction with mass spectrometry (MS) for the identification of significant differentially expressed proteins between 15cases of paired clear cell renal cell carcinoma (ccRCC) and adjacent normal renal tissues. The protein spots were considered as differentially expressed if a 1.5-fold altered expression level was observed (Student's t test, P value<0.05). Results Of the 27 differentially expressed protein spots, 26 proteins were successfully identified. 11 proteins up-regulated in renal cell carcinoma,15 proteins down-regulated. Among them Short/branched chain specific acyl-CoA dehydrogenase, mitochondrial (ACDSB), Aldose 1-epimerase (GALM), Peroxiredoxin-4 (PRDX4), Macrophage-capping protein (CAPG), Beta-defensin 107 (D107A), Microfibril-associated glycoprotein 4 (MFAP4) were first time screening as new differential expressed proteins by protomic study in renal cell carcinoma. Conclusions 2-D DIGE is a useful technique for screening and analysis differential expressed proteins in renal cell carcinoma. These new differently expressed proteins may be useful for development new molecular markers for the tumor.