The long term goal of our research project is to elucidate the mechanism(s) that regulate alpha-fetoprotein (AFP) gene expression in liver during development and hepatocarcinogenesis. We will continue and extend our previous work in this application with an emphasis on DNA sequence-specific regulation of AFP gene expression. The project includes studies to localize and characterize the regulatory sequences of the AFP gene (including enhancer and silencer sequences) by transient expression assays with various AFP-CAT fusion gene constructs. After the cis-elements are clearly defined, the next goal will be to identify the regulatory sequence-binding proteins (trans-acting factors) in various AFP-producing and non-producing cells by gel mobility shift and DNase I- footprinting assays. The purification of trans-acting factors will be done by conventional column chromatography and DNA-affinity chromatography. The project will also include studies to characterize the purified trans-acting factors and to examine chemically and physically the interaction between DNA and proteins. An in vitro transcription system will be developed from hepatoma cells that will permit further characterization of transcription factors. The main objective of this application is not only to localize and characterize the regulatory sequences and transcription factors necessary for efficient (basal) transcription of the AFP gene, but also, importantly, to determine the cis- and trans-acting elements mediating the different actions of glucocorticoid hormones on AFP gene expression in hepatoma 7777 and McA-RH8994 cells.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
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Pathology B Study Section (PTHB)
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University of Vermont & St Agric College
Schools of Medicine
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Chen, H; Dong, J M; Liu, Y et al. (1999) Identification of a cis-acting element in the rat alpha-fetoprotein gene and its specific binding proteins in F9 cells during retinoic acid-induced differentiation. J Cell Biochem 72:25-34
Liu, Y; Chen, H; Chiu, J F (1994) Identification of a retinoic acid response element upstream of the rat alpha-fetoprotein gene. Mol Cell Endocrinol 103:149-56
Fukuda, K; Kojiro, M; Chiu, J F (1994) Differential regulation of tissue transglutaminase in rat hepatoma cell lines McA-RH7777 and McA-RH8994: relation to growth rate and cell death. J Cell Biochem 54:67-77
Liu, Y; Chiu, J F (1994) Transactivation and repression of the alpha-fetoprotein gene promoter by retinoid X receptor and chicken ovalbumin upstream promoter transcription factor. Nucleic Acids Res 22:1079-86
Liu, Y; Chen, H; Dong, J M et al. (1994) cis-acting elements in 5'-flanking region of rat alpha-fetoprotein mediating retinoic acid responsiveness. Biochem Biophys Res Commun 205:700-5
Fukuda, K; Kojiro, M; Chiu, J F (1993) Cross-linked cytokeratin polypeptides in liver and hepatoma cells: possible association with the process of cell degeneration and death. Hepatology 17:118-24
Fukuda, K; Kojiro, M; Chiu, J F (1993) Demonstration of extensive chromatin cleavage in transplanted Morris hepatoma 7777 tissue: apoptosis or necrosis? Am J Pathol 142:935-46
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Zhang, X K; Zucker, M L; Huang, D P et al. (1991) Alteration of cellular oncogene expression in L1210 cells by a nitrosourea analog of thymidine. Cancer Commun 3:119-26

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