RETINOIC ACID RECEPTOR ALPHA-FETOPROTEIN AND HEPATOMA

Project: Research project

Description

The broad, long term objective of this proposal is to understand the
potential role of retinoic acid (RA) in the treatment of hepatoma. The
specific aims of the proposed experiments are to: 1) study how RA
regulates the phenotype of hepatoma cells, 2) determine the manner in
which alpha-fetoprotein (AFP) and albumin (A1b) genes are regulated during
the phenotypical change, 3) assess the regulatory mechanisms, and 4)
elucidate the roles of retinoic acid receptor (RAR) in the regulatory
process, and during hepatocarcinogenesis. Our preliminary data indicated that in Morris hepatoma McA-RH 8994 cell
line, RA enhanced the expression of AFP and A1b genes which are
characteristic of hepatocyte differentiation. To understand the involved
regulatory mechanism, the AFP mRNA half life and transcriptional rate of
AFP gene will be examined by pulse chase analysis of 3H uridine
incorporation and nuclear run off assay in RA treated or untreated McA-RH
8994 cells. The functional elements of AFP gene will be examined by
transient transfection assay using various kinds of AFP-chloramphenicol
acetyltransferase (AFP-CAT) constructs. The 5'-flanking rat AFP genomic
elements implicating RA responsiveness will be determined. Gel shift assay
will be employed to examine the in vitro binding property of the element
and the presence of binding proteins. RARbeta gene was inactivated and
resistant to RA in Morris hepatoma McA-RH 8994 and 7777 cell lines. This
aberrant expression of a normal gene may relate to carcinogenesis and
provides a model system for studying the role of RARbeta in
transformation. The cis-acting element within the promoter region of
RARbeta gene will be isolated from genomic libraries of normal rat liver
and hepatoma cells. The RARbeta gene sequence will be examined and the
presence of mutation will be identified. The trans-acting factors which
are putatively involved in control of RARbeta gene expression will also be
studied by gel shift assay. The results of this study will help us in
understanding the roles of RAR in regulating liver specific gene expression
and in hepatocarcinogenesis.
StatusFinished
Effective start/end date8/15/911/31/16

Funding

  • National Institutes of Health
  • National Institutes of Health: $274,092.00
  • National Institutes of Health: $300,235.00
  • National Institutes of Health: $291,518.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $329,615.00
  • National Institutes of Health
  • National Institutes of Health: $319,727.00
  • National Institutes of Health: $308,558.00
  • National Institutes of Health: $318,405.00
  • National Institutes of Health: $313,611.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $328,253.00
  • National Institutes of Health: $313,611.00
  • National Institutes of Health: $330,750.00
  • National Institutes of Health: $68,325.00
  • National Institutes of Health
  • National Institutes of Health: $322,978.00
  • National Institutes of Health: $309,186.00
  • National Institutes of Health
  • National Institutes of Health: $330,750.00

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Retinoids
alpha-Fetoproteins
Tretinoin
Hepatocellular Carcinoma
Retinoic Acid Receptors
Cytoplasmic and Nuclear Receptors
Genes
Hepatocytes
Xenobiotics
Liver
Liver Regeneration
Neoplasms
Down-Regulation
Gels
Retinoic Acid Receptor alpha
Transfection
Experimental Liver Neoplasms
Trans-Activators
Apoptosis
Ligands

ASJC

  • Medicine(all)