肾母细胞瘤血清非炎症标记物SAA1和载脂蛋白C-Ⅲ的筛选、鉴定及验证  

Identification of APO C-Ⅲ and SAA1 as potential serum biomarkers for nephroblastoma

作　　者：庞艳丽[1] 王磊[1] 赵伟[1] 张大[1] 秦攀[1] 樊玉霞[1] 张俊杰[1] 郭飞[1] 王家祥[1] 

机构地区：[1]郑州大学第一附属医院外科,450052

出　　处：《中华小儿外科杂志》2013年第6期424-429,共6页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金项目（编号：81172085）

摘　　要：【摘要】目的在排除炎症因子或炎性蛋白干扰的情况下，筛选肾母细胞瘤（wilmstumor，WT）血清特异性标志物，进一步鉴定与验证，提高肾母细胞瘤血清特异性标记物的敏感度与准确度。方法收集2006年1月至2010年8月郑州大学第一附属医院小儿科收治儿童的303例血清样本，WT患儿术前血清103例（术前组），WT患儿术后血清97例（术后组），健康儿童体检血清103例（对照组）。应用表面增强激光解吸电离飞行时间质谱（SELDI—TOF_MS）在排除机体常见炎症因子干扰的前提下，对术前组、术后组和对照组进行筛选并获得特异性蛋白质标记物，高效液相色谱技术（HPLC）和基质辅助激光解吸电离飞行时间质谱技术（MALDI-TOF_MS）纯化目标血清蛋白，二维液相色谱一线性离子阱质谱联用系统（2D-LC-LTO-MS）鉴定目标血清蛋白，并通过蛋白质印渍技术（Western-blot）对于以上筛选及鉴定的特异性或者差异性蛋白质进行验证。结果发现两个质荷比（m／z）为11526Da和4756Da的差异性蛋白质，均在术前组中高表达，余两组低表达，术前组与术后组、术前组与对照组比较差异有统计学意义（P均〈0．05）。经鉴定此两种差异性蛋白质为血清淀粉样蛋白A1（serumamyloidA1，SAAl）和载脂蛋白C-Ⅲ。结论本研究获得特异性更强的肾母细胞瘤非炎症性蛋白质标记物SAAl和载脂蛋白C-Ⅲ，并验证了通过这一实验思路和技术流程来鉴定肿瘤蛋白质标记物的可行性。Objective To evaluate the sensitivity and accuracy of nephroblastoma serum specific markers. Methods The serum specific markers of nephroblastoma were screened and identified based on the elimination of interference of inflammatory factor and cytokines. SELDI-TOF MS was applied to screen the serum specific markers of nephroblastoma in before and after operation and normal con trol sera. The interference of inflammatory factor and inflammatory proteins in the sera was eliminatecl before testing. The target serum proteins were purified by HPLC and MALDI-TOF MS and were i dentified by 2D-LC-LTQ-MS, followed by western blotting analysis. Results By comparing protein peaks in the sera of three groups, we identified two peaks（11 526 and 4756 Da）, with significant differences both between pre-surgery and control sera, and between pre- and post-operative sera （both, P〈0. 05）. These two peaks were further identified as serum amyloid A1 （SAA1） and apoli poprotein C-Ⅲ （APO C-Ⅲ）. Conclusions These results suggest that SAA1 and APO C-Ⅲare two potential biomarkers for nephroblastoma.

关 键 词：蛋白质标记物 血清淀粉样蛋白A1 载脂蛋白C-Ⅲ 炎症因子 肾母细胞瘤 

分 类 号：R737.11[医药卫生—肿瘤]