Cellular level of beta-galactoside alpha2,6-sialyltransferase in hepatocellular carcinoma and its role in the formation of tumor specific alpha-fetoprotein isoforms.

Abstract

Chiu Hoi Shan Clarissa.Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.Includes bibliographical references (leaves 110-126).Abstracts in English and Chinese.Abstract in English --- p.iAbstract in Chinese --- p.iiiAcknowledgement --- p.ivList of Abbreviations --- p.vList of Tables --- p.viList of Figures --- p.viiiIntroduction and ObjectivesHepatocellular CarcinomaEpidemiology --- p.1Sex and Age --- p.1Geographic distibution --- p.2Risk factors of HCC --- p.2Mortality from liver cancer --- p.4Treatment for HCC --- p.4Tumor markers --- p.5Alpha-fetoproteinStructure --- p.5Physiological Functions of AFP --- p.7Re-expression of AFP in Adult --- p.7Re-expression of AFP in HCC --- p.8Isoforms of AFP --- p.8Specific AFP isoform in HCC --- p.9Sialic Acid --- p.11Sialyltransferase"Galβ 1,4GlcnAc α2,6-sialyitransferase" --- p.12"Characterization of ST2,6Gal I" --- p.12"Expression of ST2,6Gal I" --- p.12"General features of ST2,6Gal I Activity" --- p.15Relationship Between AFP isoforms and ST2，6Gal in Fetal Mouse Model --- p.15"Change in ST2,6GaI I Activity in Transgenic Mouse Models of HCC" --- p.16"Study of Activity of ST2,6Gal I in Colon Carcinoma" --- p.16Objective of the Project --- p.18Chapter Chapter 1 --- Formation of Monosialyated AFP by Hepatoma CellsChapter 1.1 --- Introduction --- p.21Chapter 1.2 --- Materials and Methods --- p.23Chapter 1.3 --- ResultsChapter 1.3.1 --- AFP obtained from cell culture --- p.34Chapter 1.3.2 --- IEF for AFP in cell culture medium --- p.34Chapter 1.3.3 --- SDS-PAGE analysis of AFP --- p.34Chapter 1.3.4 --- Stability of AFP isoforms after secreted to cell culture medium --- p.39Chapter 1.3.5 --- Comparison of the AFP isoforms between liver tissues and serum --- p.38Chapter 1.4 --- DiscussionChapter 1.4.1 --- Origin of the extracellular msAFP - in vitro Model --- p.43Chapter 1.4.2 --- Origin of circulating msAFP - in vivo Model --- p.44Chapter 1.5 --- Conclusion --- p.46Chapter Chapter 2 --- "Presence of msAFP in the serum of HCC patient is a Consequence of Decrease in the Activity of ST2,Chapter 2.1 --- Introduction --- p.47Chapter 2.2 --- Materials and Methods --- p.49Chapter 2.3 --- ResultsChapter 2.3.1 --- Semi-quantitation of msAFP --- p.61Chapter 2.3.2 --- Evaluation of the ST2，6Gal I Assay --- p.65Chapter 2.3.3 --- "Measurements and comparisons of the activity of ST2,6Gal I in non-tumor and tumor tissue" --- p.65Chapter 2.3.4 --- "Evaluation of the RT-PCR ELISA for semi-quantitation and comparisons of ST2,6Gal I mRNA levels" --- p.75Chapter 2.3.5 --- "Semi-quantitation and comparisons of ST2,6Gal I mRNA levels in the non-tumor and tumor tissues" --- p.84Chapter 2.3.6 --- Correlations between the markers --- p.87Chapter 2.4 --- DiscussionChapter 2.4.1 --- "Overproduction of AFP, a possible cause for increased msAFP formation" --- p.99Chapter 2.4.2 --- "Decrease of ST2,Gal I activity, a possible cause for increased msAFP formation" --- p.100Chapter 2.4.3 --- "ST2,6Gal I activity in tumor is not regulated at transcriptional level" --- p.102General DiscussionOrigin of blood stream msAFP --- p.103Physiological Mechanism for the formation of msAFP in HCC --- p.104Regulation of ST2，6Gal I activity in HCC --- p.105"Comparison between the ST2,6GaI I activities in human HCC and Colon Cancer" --- p.107Conclusion and Future studiesConclusion --- p.108Future studies --- p.109References --- p.11