Differential Proteomic Analysis of Acute and Chronic Hepatitis C Virus Infections Using iTRAQ-based Technology

• Main Authors: Yu-Wei Hsu, 徐羽薇
• Other Authors: Sung-Fang Chen
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/48191560925145476259

碩士 === 國立臺灣師範大學 === 化學系 === 102 === Hepatocellular carcinoma (HCC) is one of the most prevalent and mortal cancer in the world, and 20% to 30% of patients with chronically hepatitis C virus lead to liver cirrhosis and liver cancer. Due to genetic variability of hepatitis C virus, the development of antiviral drugs and vaccines becomes a real challenge. In this study, naïve Huh7.5-SEAP cells were established and infected by hepatitis C virus, then isobaric tags for relative and absolute quantitation (iTRAQ) was applied to investigate protein profiles in both acute and chronic infections. The iTRAQ labeled peptides were fractionated by solution isoelectric focusing (sIEF) or hydrophilic interaction liquid chromatography (HILIC), followed by reversed phase nano-LC tandem mass spectrometry analysis. A total of 2615 proteins were identified, and 1816 of them were also quantified. Two-dimensional liquid chromatography technique that employed on iTRAQ labeled peptides provided results with excellent complementarity and orthogonality. Moreover, 78 and 140 differentially expressed proteins were selected in acute and chronic infection cases for GeneGo analysis. As a result, these proteins were found to be associated with cytoskeleton remodeling and cell adhesion. Besides, proteins correlated with insulin resistance, vesicular traffic, actin remodeling and secretory pathway, will be further validated by Western blot, and could serve as novel diagnostic biomarkers for hepatitis C virus infection.