The proteomics of the differential protein expressions in human glioma cell line U251 cells infected with human cytomegalovirus(HCMV)was investigated at the protein level by using the surface enhanced laser desorption...The proteomics of the differential protein expressions in human glioma cell line U251 cells infected with human cytomegalovirus(HCMV)was investigated at the protein level by using the surface enhanced laser desorption/ionization(SELDI)protein chip system in order to develop a method of study for the pathogenesis of HCMV infection.In this study,the cultured U251 cells were infected with HC- MV in good condition and the supernatants of lysates and the extracellular fluids of the cultivated infect- ed cells were quantitatively defined for the expressed proteins.The proteomics of the differential protein expression in cells before and after infection was analyzed by WCX2 arrays on the protein chip reader. It was demonstrated that the cytopathic effects of infected cells appeared on the 5th day after infection, however,the differential protein expression was evident at 6 h after infection as revealed by RT-PCR and mass spectrometry.The protein peaks captured from different batches of samples,from the same sample detected with different arrays or for the different limes were all equivalent.With the molecular weight range from 2000 Da to 3000 Da,chip captured 82 peaks from the intracellular fluids and 11 protein peak from the cellular fluid in which compared with the control group,the protein peaks with molecular weight of 13 536.3 Da,10 046.1 Da and 17 106.2 Da were close to those ofβ-amyloid pro- tein,caspase-1 precursor and LPS-induced TNF-αfactor respectively,which showed brief up-regulation 4 h after infection,and continued to raise 48 h later.These results infer that these proteins may be re- lated to the apoptosis induced by HCMV infection,thus suggesting that the apeptosis induced by HC- MV infection may play a role in the pathogenesis of HCMV infection.展开更多
目的:采用SELDI技术筛选肺癌患者的血清标志物。方法:用WCX2芯片SELDI技术分别检测31例肺癌患者、32例健康人血清、29例肺部良性病变。应用Proteinchipsoft ware 3.2软件采集蛋白峰,找出差异蛋白;应用Biomarker Pattern软件建立诊断模...目的:采用SELDI技术筛选肺癌患者的血清标志物。方法:用WCX2芯片SELDI技术分别检测31例肺癌患者、32例健康人血清、29例肺部良性病变。应用Proteinchipsoft ware 3.2软件采集蛋白峰,找出差异蛋白;应用Biomarker Pattern软件建立诊断模型。结果:肺癌患者和健康人血液标本比较,两组间有23个差异蛋白(P<0.05),用这些差异蛋白构建的诊断模型诊断肺癌敏感度为100%,特异度为96.9%,ROC曲线下面积为100%。肺癌和肺部良性病变患者的血清进行差异蛋白质组学研究发现,两组间有8个差异蛋白(P<0.05),用这些差异蛋白构建的诊断模型诊断肺癌敏感度为71.0%,特异度为62.1%,ROC曲线下面积为92.68%。结论:SELDI技术可以将肺癌患者、肺部良性病变患者和健康人正确区分,这对肺癌的筛查和早期诊断有重要意义。展开更多
基金This work was supported by National Natural Science Foundation of China(No.30471527 and No.30540075)partly supported by Mr.Tai Scholar Construction Engineering Foundation.
文摘The proteomics of the differential protein expressions in human glioma cell line U251 cells infected with human cytomegalovirus(HCMV)was investigated at the protein level by using the surface enhanced laser desorption/ionization(SELDI)protein chip system in order to develop a method of study for the pathogenesis of HCMV infection.In this study,the cultured U251 cells were infected with HC- MV in good condition and the supernatants of lysates and the extracellular fluids of the cultivated infect- ed cells were quantitatively defined for the expressed proteins.The proteomics of the differential protein expression in cells before and after infection was analyzed by WCX2 arrays on the protein chip reader. It was demonstrated that the cytopathic effects of infected cells appeared on the 5th day after infection, however,the differential protein expression was evident at 6 h after infection as revealed by RT-PCR and mass spectrometry.The protein peaks captured from different batches of samples,from the same sample detected with different arrays or for the different limes were all equivalent.With the molecular weight range from 2000 Da to 3000 Da,chip captured 82 peaks from the intracellular fluids and 11 protein peak from the cellular fluid in which compared with the control group,the protein peaks with molecular weight of 13 536.3 Da,10 046.1 Da and 17 106.2 Da were close to those ofβ-amyloid pro- tein,caspase-1 precursor and LPS-induced TNF-αfactor respectively,which showed brief up-regulation 4 h after infection,and continued to raise 48 h later.These results infer that these proteins may be re- lated to the apoptosis induced by HCMV infection,thus suggesting that the apeptosis induced by HC- MV infection may play a role in the pathogenesis of HCMV infection.
文摘目的:采用SELDI技术筛选肺癌患者的血清标志物。方法:用WCX2芯片SELDI技术分别检测31例肺癌患者、32例健康人血清、29例肺部良性病变。应用Proteinchipsoft ware 3.2软件采集蛋白峰,找出差异蛋白;应用Biomarker Pattern软件建立诊断模型。结果:肺癌患者和健康人血液标本比较,两组间有23个差异蛋白(P<0.05),用这些差异蛋白构建的诊断模型诊断肺癌敏感度为100%,特异度为96.9%,ROC曲线下面积为100%。肺癌和肺部良性病变患者的血清进行差异蛋白质组学研究发现,两组间有8个差异蛋白(P<0.05),用这些差异蛋白构建的诊断模型诊断肺癌敏感度为71.0%,特异度为62.1%,ROC曲线下面积为92.68%。结论:SELDI技术可以将肺癌患者、肺部良性病变患者和健康人正确区分,这对肺癌的筛查和早期诊断有重要意义。
