基于蛋白质组学技术筛选唐氏妊娠母体血清蛋白标志物的研究  

Prenatal maternal serum screening for Down's syndrome based on proteomic techniques

作　　者：姚远[1] 张波[1] 蒋滢[2] 罗娟[1] 吴玉娇[1] 姚宏[3] 

机构地区：[1]第三军医大学西南医院:检验科全军检验医学中心,重庆400038 [2]解放军第309医院检验科,北京100091 [3]第三军医大学西南医院:检验科妇产科产前诊断中心,重庆400038

出　　处：《第三军医大学学报》2015年第5期464-469,共6页Journal of Third Military Medical University

基　　金：重庆市科技攻关项目(CSTC2012gg-yyjs10046);第三军医大学成果转化基金(2013XZH04)~~

摘　　要：目的筛选唐氏(DS)妊娠母体血清蛋白标志物,初步验证其临床应用价值。方法收集确诊为DS妊娠的母体血清(DS组)和正常孕妇血清标本(对照组)各6例(均为16孕周采集),经二维凝胶电泳(2-DE)获得2组血清蛋白电泳图像,比较2组间蛋白含量变化,选取差异显著的蛋白质点进行生物质谱分析与鉴定,并采用Western blot方法验证筛选获得的血清蛋白标志物在DS妊娠母体血清中的变化情况(n=12)。结果通过2-DE图谱分析,共筛选出29个表达显著差异的血清蛋白质点。从中选取6个表达差异较大的蛋白质点,经生物质谱技术分析和NCBI数据库搜索与匹配,发现CFHR1和Kininogen 1等10个蛋白质与其匹配的可能性较大(匹配分值>66分)。Western blot结果显示,CFHR1和Kininogen 1在DS组的积分光密度值分别为(5 687.0±727.8)和(133 451.8±32 118.8),均明显高于对照组水平[(1 414.1±715.6)(P<0.01)和(33 073.9±14 390.4)(P<0.05)]。结论2-DE和生物质谱分析是筛选蛋白质标志物的可靠方法,Western blot结果证实CFHR1和Kininogen 1的异常变化可能与DS妊娠密切相关,有望成为DS早期产前筛查的母体血清蛋白新标志物。Objective To screen serum biomarkers of Down＇s syndrome（ DS） in the maternal serum and evaluate their values in clinical practice. Methods Maternal serum samples from 6 pregnant women who were confirmed to be carrying DS fetuses（ DS group） and 6 normal pregnant women with non-DS pregnancies（ control group） in the second-trimester were collected in our prenatal diagnostic center. The samples were analyzed by 2-dimensional gel electrophoresis（ 2-DE） to screen the proteins with different expression for potential predictive biomarkers of DS-affected pregnancies. Then the serum proteins expressed most differentially in DS group were identified by matrix assisted laser desorption / ionization time-of-flight mass spectrometry（ MALDI-TOF-MS）. Differential levels of complement factor H-related protein 1（ CFHR1） and kininogen 1 were further verified by Western blotting. Results The analysis of 2-DE results revealed 29 protein spots having significantly differed expression levels between DS and control groups. Then several differentially expressed proteins,including CFHR1 and kininogen 1,were successfully identified by MALDITOF-MS. Western blotting indicated the integrated density values of CFHR1 and kininogen 1 were 5 687. 0 ±727. 8 and 133 451. 8 ± 32 118. 8 respectively in DS group,significantly higher than those in control group（ 1 414. 1 ± 715. 6,P 0. 01,33 073. 9 ± 14 390. 4,P 0. 05）. Conclusion 2-DE and mass spectrometry analysis is a reliable method for screening protein markers. Western blotting confirms that the differential expression of CFHR1 and kininogen 1 are involved in DS-affected pregnancies,and might serve as potential predictive biomarkers for prenatal screening.

关 键 词：唐氏综合征 CFHR1 KININOGEN 1 母体血清 产前筛查 

分 类 号：R446.112[医药卫生—诊断学] R596.104[医药卫生—临床医学]