摘要
目的采用血清蛋白质组学方法筛选能诱发机体产生自身抗体的鼻咽癌（NPC）相关抗原，以便为NPC诊断和治疗提供候选标志。方法收集19份NPC组织，采用二维凝胶电泳（2-DE）技术分离组织总蛋白质，每份组织样本同时做3块2-DE胶，其中将1块2-DE胶作为制备胶进行考马斯亮蓝染色，另2块2-DE胶采用半干式电转法将胶中的蛋白转至PVDF膜上，并分别与NPC患者自身的血清和健康人血清进行2-DE免疫印迹分析，识别能与多数NPC患者血清反应而不与或很少与健康人血清反应的蛋白质点，再从制备2-DE胶中切取相应蛋白质点，进行基质辅助激光解吸电离飞行时间质谱（MOLDI—TOFMS）和电喷雾电离四级杆飞行时间质谱（ESI-Q—TOFMS／MS）分析，获取肽质量指纹图和肽序列标签，数据库搜索鉴定蛋白质。结果鉴定了13个能诱发机体产生自身抗体的NPC相关抗原：热休克蛋白70（HSP70）、人血清白蛋白（HAS）、热休克蛋白60（HSP60）、细胞角蛋白15（CK15）、亮氨酸氨基肽酶（LAP3）、α-烯醇化酶（Ot—enolase）、ErbB3结合蛋白（EBP1）、细胞角蛋白19（CK19）、核糖体蛋白P0（RPP0）、丙酮酸脱氢酶E1（PDE1）、GTP结合调节蛋白（GNBP）、抗增殖蛋白（prohibitin）和RhoGDP解离抑制因子2（Rho—GDI2），13种蛋白质点在21％的鼻咽癌患者中可见，而健康人中缺乏或出现频率较低。结论本研究筛选到13个NPC相关抗原，这些抗原能诱导患者产生自身抗体，这些抗体有可能成为NPC诊断标志和治疗靶标。
Objective To identify proteins eliciting a humoral immune response in patients with nasopharyngeal carcinoma （NPC） by a serological proteome analysis, and provide candidate biomarkers for diagnosis and treatment of NPC. Methods Two-dimensional （2-DE） elcctrophoresis was used to separate total cellular proteins from 19 NPC tissues. Separated proteins were transferred onto PVDF membranes and sera from 19 NPC patients and 19 healthy subjects were individually screened by western blotting for antibodies that react against separated proteins. Each tissue samples was subjected to three 2-DE gels and coomassie staining was performed in one of them. The protein spots which selectively reacted with the patient sera were excised from the preparative gels and subjected to further analysis of MOLDI-TOF MS and ESI-Q- TOF MS/MS. The proteins were identified based on peptide mass fingerprints and peptide sequence tags followed by searching database. Results In this study, 13 NPC associated antigens （HSP 70, HAS, HSP 60, CK 15, LAP 3, α-enolase, EBP 1, CK 19, ribosomal protein P 0, pyruvate dehydrogenase E1, guanine nucleotide-binding protein, prohibitin, Rho-GDI 2） that elicited an antibody response in most of NPC patients were identified. The positivities of these proteins were more than 21% all in NPC patients, but were lower or even absent in normal subjects. Conclusion These 13 NPC associated antigens and their autoantibodies may be useful for NPC diagnosis and treatment.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2009年第2期142-147,共6页
Chinese Journal of Laboratory Medicine
基金
基金项目：国家973重点基础研究发展规划项目资助课题（2001CB5120）
湖南省“芙蓉学者”特聘教授科学研究基金资助课题（湘教通[2007]362号）
教育部跨世纪优秀人才培养计划基金资助项目（教计函[2002]48）
湖南省科技厅重点项目资助课题（06SK2004）
关键词
鼻咽肿瘤
蛋白质组学
抗原
肿瘤
自身抗体
肿瘤标记
生物学
Nasopharyngeal neoplasms
Proteomics
Antigens, neoplasm
Autoantibodies
Tumor markers, biological
