经飞行质谱筛选门静脉癌栓血清蛋白标记物及纯化鉴定的研究

Purification and identification of serum protein biomarkers with relation to portal vein tumor thrombi screened by SELDI-TOF MS

目的筛选肝细胞癌门静脉癌栓相关的血清蛋白标记物。方法收集肝癌无癌栓患者和肝癌门静脉癌栓患者血清各12例,以弱阳离子交换蛋白质芯片为检测介质,利用表面加强激光解吸电离-飞行时间质谱测定出蛋白质谱,BioMarker Wizard软件筛选出差异蛋白峰,选择其中一个蛋白进行分离纯化和富集,利用基质辅助激光解吸飞行时间质谱对其进行鉴定。结果在m/z 1100～30000范围内,检测出的100个蛋白峰中7个差异有统计学意义(P<0.05),m/z为3397的蛋白峰在肝细胞癌伴门静脉癌栓组中上调,而m/z为7546、7896、8682、15 104、15 857和16 457的蛋白峰在肝细胞癌伴门静脉癌栓组中下调;m/z为8682的差异蛋白经鉴定为载脂蛋白A- I,与无癌栓组比较,门静脉癌栓组中该蛋白表达降低。结论筛选出的蛋白分子标记物可能与肝细胞癌门静脉癌栓形成有关,有可能为肝癌和门静脉癌栓早期预测及治疗监测提供参考。

Objective To screen serum protein biomarkers with relation to portal vein tumor thrombi （PVTT） in hepatocellular carcinoma （HCC） patients. Methods Sera of 2 groups were selected from 12 HCC patients without PVTT and 12 HCC patients with PVTT, respectively. Special serum protein or peptide pattern was determined by SELDI-TOF-MS measurement after treating the sample onto CM10 protein chip for each case. The obtained data were analyzed by BioMarker Wizard software to screen serum protein biomarkers with relation to PV＇IT. One of these protein biomarkers was purified and enriched, then it was identified by MALDI-TOF-MS. Results Ranging from 1100 to 30 000 at the m/z value, 100 protein features were detected in the serum protein pattern stably. Among them, 1 protein peak with the m/z value of 3397 was up-regulated, and 6 proteins peaks with the-m/z value of 7546,7896,9415,8682,15 104, 15 857 and 16 457 were down-regulated respectively in the group of HCC with PVTT. One of the 7 candidate protein peaks with the m/z value of 8682 was identified to be apolipoprote.in A-I. Compared with non- PVTT group, apolipoprotein A-I was decreased in PVTT group. Conclusion These candidate protein biomarkers may be related to PVTT in HCC patients and may have great clinical significance in prediction and surveillance of treatment for HCC and PVTT.