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Proteome analysis of a human liver carcinoma cell line stably expressing hepatitis delta virus ribonucleoproteins

Proteome analysis of a human liver carcinoma cell line stably expressing hepatitis delta virus ribonucleoproteins
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.
2-DE, hepatitis D virus, liver, proteome
1874-3919
616-627
Mota, Sérgio
fe39404b-e413-4834-97c6-0f2204b500a9
Mendes, Marta
47256ab9-eae2-4f3d-85e4-d214556963e2
Freitas, Natália
98eaf743-ac20-49e1-8fed-7db1b7e78b8a
Penque, Deborah
c9d54f80-1ab8-4ed1-901c-87afe53d234e
Coelho, Ana V.
fb3a7d2c-4ff6-4de0-bd55-2e7e9398be94
Cunha, Celso
21d33b16-171b-4528-9ffa-0a4c87870426
Mota, Sérgio
fe39404b-e413-4834-97c6-0f2204b500a9
Mendes, Marta
47256ab9-eae2-4f3d-85e4-d214556963e2
Freitas, Natália
98eaf743-ac20-49e1-8fed-7db1b7e78b8a
Penque, Deborah
c9d54f80-1ab8-4ed1-901c-87afe53d234e
Coelho, Ana V.
fb3a7d2c-4ff6-4de0-bd55-2e7e9398be94
Cunha, Celso
21d33b16-171b-4528-9ffa-0a4c87870426

Mota, Sérgio, Mendes, Marta, Freitas, Natália, Penque, Deborah, Coelho, Ana V. and Cunha, Celso (2009) Proteome analysis of a human liver carcinoma cell line stably expressing hepatitis delta virus ribonucleoproteins. Journal of Proteomics, 72 (4), 616-627. (doi:10.1016/j.jprot.2008.12.003). (PMID:19136081)

Record type: Article

Abstract

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.

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More information

Accepted/In Press date: 4 December 2008
Published date: 2 May 2009
Keywords: 2-DE, hepatitis D virus, liver, proteome
Organisations: Faculty of Natural and Environmental Sciences

Identifiers

Local EPrints ID: 384633
URI: http://eprints.soton.ac.uk/id/eprint/384633
ISSN: 1874-3919
PURE UUID: df20cf23-66b9-46d3-9b66-2283e6a2affa
ORCID for Sérgio Mota: ORCID iD orcid.org/0000-0002-8127-5246

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Date deposited: 03 Dec 2015 15:19
Last modified: 14 Mar 2024 22:02

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Contributors

Author: Sérgio Mota ORCID iD
Author: Marta Mendes
Author: Natália Freitas
Author: Deborah Penque
Author: Ana V. Coelho
Author: Celso Cunha

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