基质辅助激光解吸电离飞行时间质谱技术筛选乙型肝炎肝硬化至肝癌过程中血清标志蛋白的初步研究

Proteomic Markers of the Patients with Liver Cirrhosis to Liver Cancer by MALDI-TOF-MS

目的应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术分析乙型肝炎肝硬化至肝癌过程中血清标志蛋白的变化,以期发现早期肝癌甚至癌前病变时的血清标志物。方法选取2010—2012年首都医科大学附属北京世纪坛医院收治的乙型肝炎肝硬化患者240例,对其进行随访,为期3年,每6个月抽血1次,保存血清。其中12例患者在随访过程中确诊为原发性肝癌,2例患者留取血样量不足,仅对10例患者的血样进行分析。对完成随访未进展为肝癌的209例乙型肝炎肝硬化患者按照性别、年龄匹配选取40例。并选取同期本院体检中心健康者的血样40例。应用MALDI-TOF-MS技术分析患者血清蛋白质组学的变化。结果应用CM10芯片共检测出109个蛋白峰,其中具有差异的蛋白质分子量分别为:4 350.8 Da、6 858.5 Da、7 571 Da、15 125 Da、15 893 Da。其中4 350.8 Da随着肝癌的发生逐渐呈上调趋势,6 858.5 Da、7 571 Da、15 125 Da、15 893 Da随着肝癌的发生呈下调趋势。结论随着肝癌的发生,分子量为4 350.8 Da的蛋白质呈逐渐上调趋势,其余6 858.5 Da、7 571 Da、15 125 Da、15 893 Da蛋白质呈逐渐下调趋势。

Objective To analyze the changes of serum marker proteins in hepatitis B patients from liver cirrhosis to hepatocellular carcinoma by using matrix assisted laser desorption ionization time-of-flight mass spectrometry（ MALDI-TOFMS）,in order to detect serum markers of early liver cancer or even precancerous lesions. Methods From 2010 to 2012,a total of 240 patients with hepatitis B cirrhosis admitted to Beijing Shijitan Hospital,Capital Medical University were enrolled. The patients were followed up for 3 years. The blood was extracted once every 6 months and the serum was preserved. Among them,12 patients were diagnosed as primary liver cancer during follow-up. The blood samples of 2 patients were insufficient and only the blood samples of 10 patients were analyzed. 40 cases were selected from 209 patients with hepatitis B cirrhosis who completed follow-up without progression to liver cancer according to sex and age. The blood samples of 40 healthy people in the physical examination center during the same period were selected. The serum proteomic changes of patients were analyzed by using MALDITOF-MS technique. Results A total of 109 protein peaks were detected by CM10 chip,Among them,the molecular weights of the proteins with significant differences were 4 350. 8 Da, 6 858. 5 Da, 7 571 Da, 15 125 Da, and 15 893 Da,respectively. And 4 350. 8 Da showed a gradually increased trend with the occurrence of liver cancer,and 6 858. 5 Da,7 571 Da,15 125 Da and 15 893 Da showed a downward trend with the occurrence of liver cancer. Conclusion With the occurrence of liver cancer,the protein with molecular weight of 4 350. 8 Da shows a gradual upward trend,while the proteins with molecular weight of 6 858. 5 Da,7 571 Da,15 125 Da,15 893 Da show a gradual downward trend.