摘要
目的胆管癌发病率不高,但恶性程度高,早期诊断困难,迄今缺乏临床诊断可用的分子标志物。实验利用SELDI技术寻找胆管癌新的血清标志物。方法对60例胆管癌、146例肺癌、65例喉癌、58例喉咽癌、49例胆管良性疾病和53例正常人血清样品进行蛋白芯片检测。对候选蛋白进行质谱鉴定,并结合免疫共沉淀和EuSA技术筛选出胆管癌新的候选标志物。结果通过Bio—markerWizard”软件分析显示13.71×10^3,13.83×10^3和13.99×10^3的蛋白峰在胆管癌患者血清样品中明显低于对照组。通过1-D胶分离、质谱鉴定和免疫沉淀分析,显示这3个差异蛋白峰为野生型甲状腺运载蛋白(nativeTTR)和它的两个变体(cysTTR和glutTTR)。ELISA和SELDI技术分析上述血清样品均发现TTRs在胆管癌血清中的表达下调。结论采用SELDI技术首次筛选并鉴定出TTRs,表明其可能作为胆管癌诊断的候选血清标志物。
Objective As SELDI-TOF-MS (Surface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) has been broadly used to screen biomarkers for a variety of diseases, the identification and validation of the revealed biomarkers requires more focused attention. Method In this paper, the serum samples from 60 cholangiocarcinoma, 146 lung cancer, 65 LGC and 58 LPC, 49 benign diseases of hepatobiliary and 53 normal individuals were analyzed by SELDI-TOF- MS. Results Among a set of proteins automatically selected as specific biomarkers by Biomarker Wizard software, three protein peaks, with molecular weights of 13.71 × 10^3, 13.83×10^3 and 13.99 × 10^3 , were found significantly decreased in cholangiocarcinoma samples. The candidate biomarkers obtained from Tricine-SDS-PAGE gel bands by matching the molecular weight with peaks on CM10 chips were identified by Mass spectrometry as the native transthyretin(native TTR), cysTTR and glutTTR. These preliminary results were further proven by immunoprecipitation using commercial TTR antibod- ies. This allowed us to re-measure the TTR levels in all the groups more simply by ELISA assay. It showed a firm consistency between ELISA and SELDI analysis. In addition, while TTR levels in cholangiocarcinoma were found to be lower than those in normal healthy controls, TTR levels in be- nign diseases of the hepatobiliary system were found to be higher than those in healthy controls. Conclusion TTR could be a biomarker that better discriminates cholangiocarcinoma patients from the benign diseases compared to other biomarkers presently available.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2011年第2期127-131,共5页
Chinese Journal of Hepatobiliary Surgery