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Title: Comparative LC-MS analysis of proteomics profile in cells incubated with S- or R-Ibuprofen and its inhibitory effect on angiogenesis induced by HBV replication
Authors: Zhang, Jianhua
Keywords: DRNTU::Engineering::Chemical engineering::Biotechnology
Issue Date: 2011
Source: Zhang, J. H. (2011). Comparative LC-MS analysis of proteomics profile in cells incubated with S- or R-ibuprofen and its inhibitory effect on angiogenesis induced by HBV replication. Doctoral thesis, Nanyang Technological University, Singapore.
Abstract: Ibuprofen is a member of the proprionic acid group of nonsteroidal anti-inflammatory drugs (NSAIDs), with the S-enantiomer being more active compared with the R-enantiomer. It has been shown to have protective effect against neuroinflammation which is linked to the pathogenesis of several neurodegenerative disorders, including Alzheimer disease. While its prophylactic effect on Alzheimer disease has been suggested, a comprehensive understanding of its mechanism of action remains unclear. Using iTRAQ-coupled two-dimensional liquid chromatography-mass spectrometry (2D LC-MS/MS) analysis, we report here the first study of protein profile of neuroblastoma cells incubated separately with the two enantiomers of ibuprofen. Three types of cellular proteins including metabolic enzymes, signaling molecules and cytoskeletal proteins displayed changes. The changes in the level of a number of enzymes, involved in fatty acid synthesis and antioxidant activity in cells incubated with the S-enantiomer, was further supported by the real-time PCR analysis. Chronic Hepatitis B Virus (HBV) carriers may develop hepatocellular carcinoma (HCC) by a wide range of mechanisms including angiogenesis. HCC progresses from a small nodule with no blood vessels to a large hypervascular tumor by going through an angiogenic switch. HBV infection and in particular hepatitis B virus x protein (HBx) have been shown to modulate angiogenesis. However, a comprehensive and coordinated mechanism in the HBV-induced angiogenesis still remains to be established. In this study, transient transfection of replicative HBV genome was carried out in rat primary hepatocytes (RPHs) as well as HepG2 cells. A cell-based HBV replication was established in both RPHs and HepG2 cells. 2D LC-MS/MS analysis was used to detect differentially expressed proteins in cells supporting HBV replication compared with those transfected with the empty vector. HBV replication induced angiogenesis was indicated by tube formation of endothelial cells cultured in condition medium from RPHs or HepG2 cells supporting HBV replication. Enzymes associated with angiogenesis, namely fumarate hydratase and tryptophanyl-tRNA synthetase, were identified by 2D LC-MS/MS analysis in HBV replicating RPHs and HepG2 cells.
DOI: 10.32657/10356/48622
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SCBE Theses

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