Proteomics of the differential protein expressions in human glioma cell line U251 cells infected with human cytomegalovirus,as demonstrated by the surface enhanced laser desorption/ionization(SELDI)protein chip system

摘要

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.