ANALYZING THE REGULATING GENES OF LIVER REGENERATION IN CHANGING DEGREE, PATTERN, TIMING AND VERIFYING THE ROLES OF SPECIFIC AND CLUSTER GENES BY cDNA MICROARRY
Date Issued
2005-07-31
Date
2005-07-31
Author(s)
賴鴻緒
DOI
922314B002276
Abstract
Although there are much controversy
on the initiation, regulation, metabolic
changes, and termination of liver
regeneration after partial hepatectomy that
well initiate proliferation of the remaining
hepatocytes, several factors, such as
hormones, growth factors, nutritional
components, and pharmacological agents,
have been demonstrated to directly or
indirectly affect liver regeneration.
However, the regenerative mechanism and
genetic control of liver after major tissue loss
is still not clear.
The regenerating liver is a system in
which the relationships between
proto-oncogene expression and cell
replication should be examined during a
physiologic growth response.
Proto-oncogene expression after partial
hepatectomy should be specific, sequential,
and highly regulated. As measured by levels
of mRNAs, the changes have been detected
in the expression of c-fos, c-myc, p53, p21,
gas-6 and the ras gene family (c-Ha-ras,
c-Ki-ras, and N-ras). In contrast, expression
of c-src and c-abl does not change after
partial hepatectomy while c-mos transcripts
cannot be detected in normal or regenerating
liver. Arora et al reported that c-Myc
antisense limits rat liver regeneration by
regulating cytochrome p-450 3A activity.
Ozeki and Tsukamoto found that retinoic
acid can repress c-fos and c-jun expression
and induce apoptosis in regenerating liver.
Our previous study monitored the variation
of regulating genes by 384 liver-related gene
cDNA microarray nylon membrane, and
found that there are 59 proto-oncogenes
expression increased markedly and 19
decreased significantly during liver
regeneration. However, the changing degree,
patterns, timing and gene grouping were very
sophisticated and not clear. Mass survey and
more detailed analysis by more cDNA
microarry method should be very important.
Male Wistar rats around 200g will be
used as subject. Partial hepatectomy around
70% were performed. They were sacrificed
before and 2, 4, 6, 12, 24, 72 hours and 5, 7,
10 days after hepatectomy. We have
measured: (1)weight of remnant liver;
(2)mitotic index; (3)genomic survey of the
gene expression by microarray of 6144
identified cDNA clones on nylon membrane
(Wittech Co., Taipei, Taiwan), labeling of
liver mRNA hybridization and image
analysis; and (4)Grouping of genes
expression into immune, nutrition, hormone,
growth factor, enzyme, oncologic and
embryonic subgroups, and compare the
expression degree, changing pattern and
specific timing.
The results were: (1) the remnant liver
weight increased to 90% in 72h after partial
hepatectomy; (2) the mitosis of hepatocytes
increased marked at 48h then decreased at 72
after partial hepatectomy; (3) analyzing the
gene expression of microarray chips, the
variation could be classified into 72 different
patterns in cluding the patterns with a single
peak at 2, 4, 6, 12, 24, 72h and 5, 7d after partial hepatectomy; (4) gene clusters of
immune, hormone, growth factor, enzyme
and angiogenesis have changed markedly; (5)
early stage changed genes including
fas-associating protein with death domain,
carnitine palmitoyltransferase 1, fas death
domain-associating protein, and steroid
O-acyltransferase 1 could be related to the
initiation of liver regeneration; (6)
intermediate stage changed genes including
transforming growth factor beta 2 and beta
receptor could be related to the
differentiation of liver regeneration; (7) late
stage changed genes including TGF-β
regulated gene 3 and small inducible
cytokine A2 could be related to the
termination of liver regeneration.
Subjects
liver regeneration
partial
hepatectomy
hepatectomy
proto-oncogene
microarray
genetic changing pattern
gene cluster
Publisher
臺北市：國立臺灣大學醫學院外科
Type
report
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