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Proteomic Analysis of Different Phenotype of Hepatocellular Carcinoma and Textual Research of S100A9

Author: ChenBin
Tutor: PengMinHao
School: Guangxi Medical University
Course: Oncology
Keywords: human hepatocellular carcinoma tumor invasion tumor recurrence two-dimensional electrophoresis matrix assisted laser desorption/ionization time of flight mass Spectrometry S100 calcium binding protein A9 (calgranulin B)
CLC: R735.7
Type: PhD thesis
Year: 2006
Downloads: 185
Quote: 0
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Abstract


Objective Using a comparative proteomic approach, we screened key proteinic molecules correlated with invasion and recurrence by comparing the difference in protein expression of different phenotype of human hepatocellular carcinoma (HCC) tissues. Methods 5 cases of invasive HCC and 5 cases recurrent HCC tissues proved by clinical and pathological examination were divided into two groups(contrasted with 5 cases non-invasive and non-recurrent HCC tissues, respectively). Totalprotein extracted from the specimens of these groups were separated by two-dimensional gel electrophoresis (2-DE). IPG drystrip (13cm, pH3~10 non-linear) was used in the first dimensional IEF electrophoresis, and 12.5% SDS-PAGE for the second dimensional vertical electrophoresis. The preparative 2-DE gels stained with Coomassie brilliant blue were scanned using ImageScanner and analyzed with ImageMaster 2D Platinum 5.0 software. Selected differential protein spots (showing 2 or more than 2 fold difference and more than 50% emerging frequency) of comparative groups were digested into peptides by trypsin in-gels. Then these peptides were assayed by MALDI-TOF MS. Data were inputted the searching engine MASCOT (Matrix Science) against the NCBI non-redundant protein sequence database and homologous protein could be identified. Validation procedures for the screened protein S100A9 were then carried out by immuno-histochemistry, reverse transcription polymerase chain reaction analysis (RT-PCR) and western blot assay. Expressions of S100A9 in different type of liver tissue specimens were observed by immunohistochemistry. The

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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