摘要
目的 探索食管鳞状细胞癌的肿瘤相关抗原.方法 提取食管鳞状细胞癌组织蛋白,用20例食管癌患者血清和20例健康对照血清分别作为检测抗体进行Western blot分析,比较分析二者的差异,将差异蛋白酶解后用质谱进行鉴定分析;用免疫组化方法对鉴定的肿瘤相关抗原进行验证.结果 食管癌血清的阳性信号比较多,其中70kDa的蛋白阳性率为40%（8/20）,而健康对照血清阳性率为0%（0/20）,该蛋白被鉴定为HSC70或称HSPA8.免疫组化结果显示HSC70在食管癌组织主要表达于细胞浆和细胞核,且表达明显高于癌旁正常组织.结论 联合免疫印记和质谱技术可以鉴定肿瘤相关抗原;HSC70可能是食管癌发生的重要参与蛋白,可能是食管癌诊断和治疗的关键靶分子.
Objective To identify novel tumor associated antigens （TAA）in esophageal squamous cell carcinoma（ESCC）. Methods The proteins extracted from tissues of ESCC were separated by two dimensional polyacrylamide gel electrophoresis, and followed by Western blot analysis in which sera from 20 patients with ES- CC and 20 healthy controls were tested for primary antibodies. Positive spots were excised from Coomassie blue - stained gels and the tryptic peptides were analyzed by matrix - assisted laser disorption/ionization time - of - flight mass spectrometry（ MALDI - TOF - MS）. The expression of the candidate TAA in ESCC tissue was detec- ted by immunohistochemical staining（IHC）. Results Sera from patients with ESCC yielded multiple spots, one of which was identified as heat shock 70 kDa protein 8 （ HSC70 or HSPA8） by＇ MALDI - TOF - MS. The immunohistochemistry analysis showed that HSC70 was located in both cytoplasm and nucleus, and had a higher expression in cancer tissues than paracarcinoma normal tissue. Conclusion The combination of immunoblot and mass spectrometry may identify some novel TAAs. These results suggested that HSCTO may play a role in tumorigenesis of ESCC, and may be a key diagnostic and therapeutic target.
出处
《实用肿瘤学杂志》
CAS
2012年第2期137-140,共4页
Practical Oncology Journal
关键词
食管鳞状细胞癌
肿瘤相关抗原
HSC70
Esophageal squamous cell carcinoma,Tumor associated antigen, HSC70
