摘要
为了寻找乳腺癌血清标志蛋白标志物,本研究用试剂盒去除血清样品中的高丰度蛋白,通过固相pH梯度双向凝胶电泳分离乳腺癌病人血清和非乳腺癌血清,银染显色,分析比较电泳图谱寻找差异蛋白.用基质辅助激光解吸电离飞行时间质谱测定差异蛋白的肽指纹图谱,MASCOT软件分析确定为何种蛋白,然后用蛋白印迹验证与乳腺癌发生关系密切的差异蛋白.本研究获得了分辨率高、重复性好的双向电泳银染图谱,得到3个差异明显的蛋白,经肽指纹分析并结合数据库查询,初步鉴定为人血凝级联反应中的关键酶、人体细胞凋亡蛋白抑制剂样蛋白ILP-2、人体HP蛋白.蛋白印迹研究表明ILP-2在乳腺癌血清中的含量明显高于非乳腺癌血清样本,该蛋白有可能是乳腺癌标志物.
Early diagnosis is a one of the key to improve the breast cancer patients＇ prognosis.To investigate the breast cancer serum-biomarkers,high-abundant protein was depleted from 400（portion） breast cancer patients＇ serum and 40 healthy control serum.The enriched proteins were then separated by 2D-PAGE（two-dimensional polyacrylamide gel electrophoresis） and the gels were dyed by silver-stain,then three different proteins were eluted from the gels.MOLDI TOF MS（matrix-assisted laser desorption ionization flying mass） analysis revealed the identity of those proteins are：① human crystal structure of the fxia catalytic domain in complex with Ecotinm84r;② human IAP-like protein 2（Inhibitor of apoptosis protein（IAP）-like protein-2（ILP-2）,and ③ haptoglobin alpha（2FS）-beta precursor（HP）.Western-blot revealed that ILP-2 has higher concentration in breast cancer patients serum compare to those control subjects and it is a hopeful serum-biomarker for diagnosis of breast cancer.This study provides a practicable approach to seek any other potential candidate as early diagnostic biomarkers in human malignant tumors
出处
《云南大学学报（自然科学版）》
CAS
CSCD
北大核心
2011年第4期479-483,共5页
Journal of Yunnan University(Natural Sciences Edition)
基金
国家自然科学基金资助项目(30360040)
云南省科技攻关项目资助(2006SG09)
赣南医学院重点科研课题(2008100)
