蛋白芯片技术对肝细胞癌组织蛋白谱的建立及标志蛋白的筛选

目的:应用表面增强激光解吸附电离飞行时间质谱技术(SELDI-TOF-MS)对肝细胞癌组织的蛋白表达谱进行分析,从中筛选出标记蛋白.方法:采用CM10芯片及SELDI-TOF-MS技术对肝细胞癌组织26例和肝硬化组织18例进行蛋白指纹图谱检测分析;比较高、中、低分化肝细胞癌组织,不同TNM分期肿瘤以及AFP阴、阳性肝细胞癌组织的蛋白表达差异,所得结果均采用Biomarker Wizard软件进行分析;通过查询蛋白库,对特定分子质量所对应的标记蛋白进行初步确定.结果:肝细胞癌和肝硬化组织蛋白表达图谱之间存在16个稳定的标志蛋白,7个蛋白在肝细胞癌组织中表达上调.9个蛋白在肝细胞癌组织中表达下调,其中4.7 kDa,7.2 kDa和9.8 kDa蛋白峰差异性最明显;中分化和高分化肝细胞癌之间存在11个标志蛋白;通过查询蛋白库ExPasy并进行筛选,初步确定特定分子质量所对应的蛋白.结论:采用SELDI-TOF-MS技术,证实在肝细胞癌组织中存在多种高度特异性低分子蛋白.

Using protein chip technology to screen for tissue proteomic profiles and tumor markers in hepatocellular carcinoma

AIM:To use the surface-enhanced laser desorption and ionization time-of-flight mass spectrometry(SELDI-TOF-MS)technique to screen for tissue biomarkers in patients with hepatocellular carcinoma(HCC),but with different clinicopathological features. METHODS:Proteomic spectra were examined and analyzed by mass spectroscopy in 44 cases, including 26 specimens of HCC tissue that had been pathologically confirmed in patients aged 34-68 years,and 18 specimens of liver cirrhosis tissue in patients aged 38-70 years.The spectra obtained were analyzed using the biomarker wizard system,and the biomarkers were defined by searching www.ExPasy.org. RESULTS:A total of 16 distinguished pro- teomic biomarkers,7 up-regulated and 9 down- regulated,were detected from screening HCC tissue,in contrast with liver cirrhosis tissue. There were significant differences in the protein peaks of different molecular masses of 4.7,7.2 and 9.8 kDa between HCC and liver cirrhosis tissues.Eleven distinguished proteomic bio- markers were screened when comparing cases of moderately and highly differentiated HCC tissue.All proteins were confirmed by searches of www.ExPasy.org. CONCLUSION:The SELDI-TOF-MS technique offers a unique platform for proteomic detec- tion in HCC.It is also a non-invasive method for studying proteomic changes in the development and progression of HCC.