Comparative proteome analysis of a human liver cell line stably transfected with hepatitis D virus full-length cDNA.
Hepatitis D virus (HDV) is the causative agent of one of the most severe forms of virus hepatitis. HDV is a satellite virus of Hepatitis B virus (HBV) and coinfects or superinfects liver cells already infected with HBV. Investigation of HDV biology and pathogenesis has been so far impaired by the lack of an appropriate cell culture system capable of replicating and propagating the virus. This is usually partially overcome using transiently transfection systems and stably transfected cell lines. Here, we used a well-characterized human liver hepatoma cell line stably transfected with HDV cDNA (Huh7-D12) to study the changes in the host proteome induced by the expression of the virus. A 2-DE and MS approach was performed allowing the identification of 23 differentially expressed proteins when compared with the parental non-transfected Huh7 cell line. The proteomic results were validated by western blot and real-time qPCR.
Unidade de Biologia Molecular, Centro de Malária e outras Doenças Tropicais, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal. ccunha@ihmt.unl.pt
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D015153:Blotting, Western; D045744:Cell Line, Tumor; D002966:Clathrin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D003698:Hepatitis Delta Virus / Q000235:genetics*; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D034904:Lamin Type A / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008099:Liver / Q000473:pathology*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010449:Peptide Mapping; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014162:Transfection; D014357:Trypsin / Q000737:chemistry
D002966:Clathrin; D018076:DNA, Complementary; D004279:DNA, Viral; D034904:Lamin Type A; D010446:Peptide Fragments; D020543:Proteome; D012333:RNA, Messenger; D014357:Trypsin