纯化浅表膀胱肿瘤蛋白质表达谱构建及尿液生物标记发现

Construction of purified proteome expression profile of superficial bladder tumor and discovery of urine biomarker

目的总体描述浅表膀胱肿瘤蛋白质表达谱的特点,在总体描述基础上探讨浅表膀胱癌尿液蛋白质芯片的发现。方法①激光捕获显微切割获得纯化的浅表膀胱肿瘤细胞。②二维液相色谱分离多肽混合物,电喷雾串联质谱和生物信息学方法分析鉴定蛋白质。结果4例浅表膀胱移行细胞癌标本中440个蛋白质共同表达。76(17.3%)个蛋白质相对分子质量(MW)<10kDa或者>100kDa,84(19.1%)个蛋白质等电点(pI)>9。312、304个蛋白质具有基因本体论生物学途径、细胞组分注解。41、22个基因本体论生物学途径术语为浓集、缺失表达。25、11个基因本体论细胞组分术语为浓集、缺失表达。基因本体论生物学途径浓集、缺失表达分析与尿液蛋白质表达谱分析一致性较强。属于细胞粘附、细胞增殖和细胞分化类别的蛋白质是尿液中检测肿瘤的良好生物标记候选蛋白。结论浅表膀胱肿瘤蛋白质表达谱研究为理解肿瘤细胞生物学及从尿液中发现生物标记提供了有价值的信息。

Objective To characterize the proteome expression overall profile of superficial bladder tumor. On the basis of expression profile,the discovery of urine biomarker was discussed. Methods①Purified highly superficial transitional cell carcinoma cells were obtained by laser capture microdissection （LCM）;②Peptide mixture were separated by two-dimensional liquid chroma-tography （2D-LC）,followed by electrospray ionization-tandem mass spectrometry （ESI-MS/MS） identification and bioinformatics analysis of the identified proteins. Results A total of 440 proteins appeared co expression in 4 specimens. There were 76 （17.3%） of proteins with molecular weight （MW）〈10 kDa or 〉 100 kDa, 84（19.1%） proteins with pI〉9,312/304 proteins with biological process/cellular component annotation as to the Gene Ontology （GO）. There were 41/22 GO terms exhibited as enriched/depleted within the biological process annotation and 25/11 GO terms exhibited as enriched/depleted within the cellular component annotation. The GO biological process enrichment/depletion analysis was strongly consistent with that of urine proteome. Proteins that belong to the term of cell adhesion, cell proliferation, cell differentiation are good candidate biomarkers for cancer detection from urine. Conclusions The present experiment identified an extensive expression profile in superficial bladder tumor, providing valuable information for the understanding of cell biology and discovery of urine biomarkers.