Proteomic analysis of host responses in HepG2 cells during dengue virus infection.
Dengue virus infection remains a public health problem worldwide. However, its pathogenic mechanisms and pathophysiology are still poorly understood. We performed proteomic analysis to evaluate early host responses (as indicated by altered proteins) in human target cells during dengue virus infection. HepG2 cells were infected with dengue virus serotype 2 (DEN-2) at multiplicity of infection (MOI) of 0.1, 0.5, and 1.0. Quantitative analyses of DEN-2 infection and cell death at 12, 24, and 48 h postinfection showed that the MOI of 1.0 with 24 h postinfection duration was the optimal condition to evaluate early host responses, as this condition provided the high %Infection ( approximately 80%), while %Cell death ( approximately 20%) was comparable to that of the mock-control cells. Proteins derived from the mock-control and DEN-2-infected cells were resolved by 2-D PAGE ( n = 5 gels for each group) and visualized by SYPRO Ruby stain. Quantitative intensity analysis revealed 17 differentially expressed proteins, which were successfully identified by peptide mass fingerprinting. Most of these altered proteins were the key factors involved in transcription and translation processes. Further functional study on these altered proteins may lead to better understanding of the pathogenic mechanisms and host responses to dengue virus infection, and also to the identification of new therapeutic targets for dengue virus infection.
Pattanakitsakul
Sa-Nga
SN
Medical Molecular Biology Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Rungrojcharoenkit
Kamonthip
K
Kanlaya
Rattiyaporn
R
Sinchaikul
Supachok
S
Noisakran
Sansanee
S
Chen
Shui-Tein
ST
Malasit
Prida
P
Thongboonkerd
Visith
V
D045744:Cell Line, Tumor; D003715:Dengue / Q000821:virology; D003716:Dengue Virus / Q000472:pathogenicity*; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology*; D006801:Humans; D040901:Proteomics