应用RP-HPLC/TOF-MS技术识别肝癌血浆差异蛋白质

Identification of differential proteins in plamsa of hepatocellular carcinoma by RP-HPLC/TOF-MS

目的应用RP-HPLC／TOF-MS技术筛选肝癌血浆差异蛋白质。方法9例肝癌和8例正常组血浆采用Agilent高丰度蛋白质分离系统（multiple affinity removal system，MARS）洗脱白蛋白等高丰度蛋白质后，样品首先采用RP—HPLC法分离分析，然后采用基质辅助激光解吸飞行时间质谱鉴定差异蛋白质相对分子质量。结果肝癌血浆和健康志愿者血浆的RP-HPLC图谱中存在3组差异蛋白质，相对分子质量为（15～50）×10^3的蛋白质在健康志愿者中有较高表达，而相对分子质量为（50～90）×10^3的蛋白质在肝癌患者中有较高表达。结论RP—HPLC／TOF-MS技术可以发现肝癌发生发展过程中血浆蛋白表达谱质或量的变化。

Objective To identify the differential proteins in plasma of hepatocellular carcinoma （HCC） with RP-HPLC and matrix-assisted laser desorption ionzation time of flight mass spectrum （ MALDITOF MS）. Methods Plasma from 8 healthy volunteers and 9 HCC patients were collected. After depleting six plasma proteins of the highest abundance by the Multiple Affinity Removal System （MARS） Technologies from Agilent, plasma were subjected to RP-HPLC and the molecular weight of differentially expressed protein was identified by MALDITOF MS. Results The proteins whose molecular weight are between 15 ×10^3 and 50×10^3 in plasma from healthy subjects were more than in hepatocelluar carcinoma patients, while the proteins whose molecular weight are between 50 ×10^3 and 90 ×10^3 in plasma from healthy subjects are less than in hepatocelluar carcinoma patients. Conclusion RP-HPLC/TOF MS based plasma proteome analysis can be useful in detecting protein expression alteration resulting from the carcinogenesis and development of HCC and new discovered biomarkers might be an aid in the diagnosis of HCC.