目的应用蛋白质组学方法筛选肺腺癌患者血清中的肿瘤标志物。方法收集8例肺腺癌患者及8例正常人血清,剔除血清中白蛋白干扰后,得到血清可溶性总蛋白并采用双向电泳方法分离,获得血清蛋白质的双向电泳图谱,经PDQuest7.1凝胶图像软件分析找出表达差异的蛋白点,经基质辅助电离解析飞行时间质谱检测获得待测蛋白的肽质量指纹图谱,通过搜寻蛋白质数据库鉴定出肺腺癌血清中候选的肿瘤相关蛋白。结果建立了肺腺癌患者及正常人血清蛋白质表达谱。经软件匹配分析,共筛选出24个差异的蛋白点,从中选出差异显著的7个蛋白点进行质谱分析,经数据库查询分析,成功的鉴定出5种蛋白质:凝聚素、触珠蛋白、血清淀粉样蛋白A、血清白蛋白A链、纤维胶凝蛋白3。结论本研究建立了人肺腺癌患者及正常人血清的双向电泳图谱;鉴定出了若干肺癌相关蛋白质,为进一步寻找肺癌蛋白标志物奠定了基础。
Objective To screen the tumor markers in serum of patients with lung adenocarcinoma by proteome method. Methods Serum samples were obtained from 8 patients lung adenocarcinoma and 8 healthy donors. After getting rid of various serum albumin, all the soluable proteins in serum were separated by two-dimensional polyac- rylamide gel electrophoresis, then the differentially expressed proteins were identified by using PDQuest software a- nalysis. Peptide mass fingerprint maps were obtained by matrix-assisted laser desorption/ionization time of flight mass spectrometry and protein identification was performed by searching protein database. Results Perfect repro- ducible two-dimensional polyacrylamide gel electrophoresis image were successfully obtained. Twenty-four differenti- ally expressed proteins were found in serum between patients with lung adenocarcinoma and healthy donors. Peptide mass fingerprinting maps of 7 proteins were obtained and 5 proteins were identified eventually, including clusterin, haptoglobin 2-alpha, amyloid related serum protein, serum albumin chain A, ficolin-3. Conclusion The two-di- mensional polyacrylamide gel electrophoresis maps of serum from patients with lung adenocarcinoma and healthy do- nor were constructed, several proteins identified in this research may be the potential serum biomarkers for lung cancer.