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Proteomic Study of HepG2 Cell Extract During Dengue Virus Infection
Rosa, M.S.1; Lery, L.M.S.3; Villas-Vôas C.S.A.2; Sa, L.A.3; Bisch, P.M.3; Da Poian, A.T.2; Lomeli, M.M.2 and Mohana-Borges, R.1
1 Laboratório de Genômica Estrutural – IBCCF – UFRJ; 2 Instituto de Bioquímica Médica – IBqM - UFRJ; 3 Unidade Multidisciplinar de Genômica – IBCCF – UFRJ
Dengue virus is one of the most important human virus pathogens transmitted by mosquitoes and causes one hundred million or more cases of infection worldwide each year, resulting in around 24000 deaths. More than 2.5 billion people are at risk of infection and more than 100 countries have endemic dengue transmission. Dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) are progressively more severe clinical manifestations of dengue infection. One of the difficulties encountered in combating the spread of DF is the existence of four different viral subtypes that do not confer cross-immunity. More alarmingly, it has been postulated that successive infections with the DF virus increase the risk of developing more severe forms of this disease, such as DHF and DSS.
Although several approaches have been tried, no commercial vaccine or treatment against DF is available. A detailed understanding of the molecular mechanisms of the interactions between the DF virus and its target cells during the infection process is essential to the development of anti-viral treatments and new therapies. An important goal is to comprehensive survey cellular proteomes and profile protein changes on cell types affected: monocytes, endothelial cells, and hepatocytes.
We employed a proteomic strategy to study the cellular proteins differentially expressed during dengue virus infection, with the aim of identifying proteins involved in each stage of the viral replication cycle. The cell model for this study was the HepG2 cell line (human, primary liver cancer) and the technologies used include protein separation of the complex mixture by two dimensional electrophoresis (2-DE) and protein identification by mass spectrometry (MS) after a trypsin digestion. The cell was infected with dengue virus subtype 2 and the protein extract was used for the analysis. We have tried to identify first cellular several proteins that appeared on the gel and some that were differentially expressed between the virus-infected and control cells. Our ultimate goal is to find out protein(s) that could be molecular markers of dengue virus infection as well as targets for an antiviral drug developed by rational drug design.
Funded by: CNPq, WHO/TDR, FAPERJ, PRONEX-RIO, IMBEB2.
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