Study On Differential Expression Proteins Of Stable Transfected Chang Liver Cell Line Expressing Hepatitis B Virus HBsAg And EGFP Fusion Protein

Hepatitis B virus (HBV) infection causes acute and chronic hepatitis. Acute infections may produce serious illness, and approximately 0.5% terminate with fatal, fulminant hepatitis. Chronic infections may also have serious consequences, HBV is considered to be a major etiological factor in the development of human hepatocellular carcinoma (HCC), one of the most frequent fatal malignancies worldwide, and worldwide deaths from HCC caused by HBV infection probably exceed one million per year. So, understanding of the precise mechanism of HBV carcinogenesis is a vital prerequisite for the development of antiviral concepts and treatment for HCC. HBV DNA integration in human genome is one of main mechanisms.HBV is a small, enveloped DNA virus of the hepadnavirus family, its genome is a relaxed-circular, partially duplex DNA of 3.2 kb, characterized by four overlapping ORFs (S,C,P,X). The S-ORF consists of a single open reading frame divided into three coding regions: pre-S1, pre-S2 and S, each starting with an in-frame ATG codon. Through alternate translational initiation at each of the three AUG codons, a large (LHBs; pre-S1 +pre-S2+S), a middle (MHBs; pre-S2+S) and a small (SHBs;S) envelope glycoprotein can be synthesized.To study on differential expression proteins of stable transfected Chang Liver cell line expressing hepatitis B virus HBsAg and EGFP fusion protein, we construct a eukaryotic expression vector for expressing hepatitis B virus (HBV)recombinant HBsAg-EGFP fusion protein and obtain a stable transfected Chang Liver cell line. Then, the total proteins of Chang-Liver-HBsAg-EGFP cell line and controll cell line were extracted and separated by 2-DE, and the images analysis were performed with ImageMaster 2D Paltinum 5.1 software. The differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-Q-TOF MS) , and a total of 9 differential expression proteins were successfully identified. There were 6 up-regulated proteins such as Protein disulfide-isomerase precursor, HSP70, stratifm, peroxiredoxin 1, Peroxiredoxin 5, guanylate-binding protein. Meanwhile , Transgelin-2, capping protein, Porin 31HM were down-regulated. Most of the differentially expressed proteins have been reported to participate in cell growth, cell damage repair and stress-reaction. The data will be helpful to illuminate the mechanisms.