Proteome analysis of a human liver carcinoma cell line stably expressing hepatitis delta virus ribonucleoproteins.
Hepatitis delta virus (HDV) infects human hepatocytes already infected with the hepatitis B virus increasing about ten fold the risk of cirrhosis and fulminant hepatitis. The lack of an appropriate cell culture system capable of supporting virus replication has so far impaired the detailed investigation of the HDV biology including the identification of host factors involved in pathogenesis. Here, we made use of a HDV cDNA stably transfected cell line, Huh7-D12, in a proteomic approach to identify the changes in the protein expression profiles in human liver cells that arise as a consequence of HDV replication. Total protein extracts from Huh7-D12 cells and of the corresponding non transfected human liver carcinoma cell line, Huh7, were separated by 2-DE. Differentially expressed spots were identified by MALDI-TOF followed by database searching. We identified 23 differentially expressed proteins of which 15 were down regulated and 8 up regulated in Huh7-D12 cells. These proteins were found to be involved in different cellular pathways. The down regulation of the histone H1-binding protein and of triosephosphate isomerase was confirmed by Real time PCR, and the up regulation of the La protein and lamin A/C was validated by western blot.
Mota
Sérgio
S
Unidade de Biologia Molecular, Centro de Malária e outras Doenças Tropicais, Instituto de Higiene e Medicina Tropical, Rua da Junqueira 96, Lisbon, Portugal.
Mendes
Marta
M
Freitas
Natália
N
Penque
Deborah
D
Coelho
Ana V
AV
Cunha
Celso
C
D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000821:virology; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism* / Q000821:virology; D020543:Proteome / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
D020543:Proteome; D012261:Ribonucleoproteins; D014764:Viral Proteins