iTRAQ定量蛋白质组学技术筛选唐氏综合征孕母血清差异蛋白  

i TRAQ-based proteomic identification of novel protein markers for Down's syndrome in maternal serum

作　　者：蒋春艳[1] 王成东[1] 张海灵[1] 王瑞丽[1] 赵桂凤[1] 

机构地区：[1]潍坊市人民医院产前诊断科,山东潍坊261041

出　　处：《哈尔滨医科大学学报》2015年第6期513-517,520,共6页Journal of Harbin Medical University

基　　金：山东省科技发展计划项目(2011YD21004);潍坊市卫生发展计划项目(20110023)

摘　　要：目的筛选唐氏综合征(Down’s syndrome,DS)母体血清差异表达蛋白质,确定和验证DS相关的候选标志物。方法收集DS胎儿(实验组)和正常胎儿(对照组)母体血清各15例,组内等体积混合后采用免疫亲和柱去除高丰度蛋白。同位素标记的绝对和相对定量技术(i TRAQ)和2D-LC-MS/MS法进行蛋白质鉴定和相对定量。ELISA法进一步验证血清中候选标志物含量。结果与对照组相比,实验组中共鉴定出有定量信息的蛋白质106个,其中差异1.5倍以上的有38个,包括26个上调蛋白和12个下调蛋白。其中血清铜蓝蛋白(ceruloplasmin,CP)上调明显,高达2.4倍。ELISA法进一步证实,实验组CP含量明显高于对照组(P<0.05)。结论基于i TRAQ技术的蛋白质组学方法能鉴定出多种DS相关的差异蛋白,CP可作为DS筛查的新标志物。Objective To identify candidate protein biomarkers by screening some differential- ly expressed proteins from maternal serum. Methods Serum samples were collected from pregnancy with and without DS fetal, including a pregnancy DS group of 15 women and a con- trol group with the same number. The samples in the same group （DS or control） were pooled to obtain equal amounts of proteins. High abundant proteins in the serum were removed by im- muno-chromatography assay. Proteins identification and relative quantification were analyzed by isobaric tags for relative and absolute quantitation （iTRAQ） coupled with 2D-LC-MS/MS. ELISA was used to confirm the levels of candidate protein biomarkers. Results A total of 106 proteins were successfully quantified, among which 38 showed at least 1.5-fold differences be- tween the DS and control group. Among 38 proteins, 26 and 12 were up- or down-regulated, respectively. Of all, eeruloplasmin（CP） , as an iron transport protein, was 2.4-fold higher（ P 〈 0.05 ） in the DS than controls. The high level of CP was also confirmed by ELISA. Conclusion A number of proteins are identified by iTRAQ-based proteomic approach and ceruloplas- min could be served as a new biomarker for DS screening.

关 键 词：唐氏综合征 母体血清 蛋白质组学 ITRAQ 

分 类 号：R446.112[医药卫生—诊断学]