|
Lun-Xiu
Qin, Zhao-You Tang, Liver Cancer Institute and Zhong shan
Hospital, Fudan University, Shanghai, China
Correspondence to: Zhao-You Tang, M.D., Professor of Surgery
& Chairman, Liver Cancer Institute & Zhongshan Hospital,
Fudan University, 136 Yi Xue Yuan Road, Shanghai 200032, China. zytang@srcap.stc.sh.cn
Telephone: +86-21-64037181 Fax: +86-21-64037181
Received 2002-03-19 Accepted 2002-05-08
Abstract
The prognosis of hepatocellular carcinoma (HCC) still remains
dismal, although many advances in its clinical study have been made.
It is important for tumor control to identify the factors that
predispose patients to death. With new discoveries in cancer
biology, the pathological and biological prognostic factors of HCC
have been studied quite extensively. Analyzing molecular markers
(biomarkers) with prognostic significance is a complementary method.
A large number of molecular factors have been shown to associate
with the invasiveness of HCC, and have potential prognostic
significance. One important aspect is the analysis of molecular
markers for the cellular malignancy phenotype. These include
alterations in DNA ploidy, cellular proliferation markers (PCNA,
Ki-67, Mcm2, MIB1, MIA, and CSE1L/CAS protein), nuclear morphology,
the p53 gene and its related molecule MDM2, other cell cycle
regulators (cyclin A, cyclin D, cyclin E, cdc2, p27, p73), oncogenes
and their receptors (such as ras, c-myc, c-fms, HGF, c-met, and erb-B
receptor family members), apoptosis related factors (Fas and FasL),
as well as telomerase activity. Another important aspect is the
analysis of molecular markers involved in the process of cancer
invasion and metastasis. Adhesion molecules (E-cadherin, catenins,
serum intercellular adhesion molecule-1, CD44 variants), proteinases
involved in the degradation of extracellular matrix (MMP-2, MMP-9,
uPA, uPAR, PAI), as well as other molecules have been regarded as
biomarkers for the malignant phenotype of HCC, and are related to
prognosis and therapeutic outcomes. Tumor angiogenesis is critical
to both the growth and metastasis of cancers including HCC, and has
drawn much attention in recent years. Many angiogenesis-related
markers, such as vascular endothelial growth factor (VEGF), basic
fibroblast growth factor (bFGF), platelet-derived endothelial cell
growth factor (PD-ECGF), thrombospondin (TSP), angiogenin,
pleiotrophin, and endostatin (ES) levels, as well as inratumor
microvessel density (MVD) have been evaluated and found to be of
prognostic significance. Body fluid (particularly blood and urinary)
testing for biomarkers is easily accessible and useful in clinical
patients. The prognostic significance of circulating DNA in plasma
or serum, and its genetic alterations in HCC are other important
trends. More attention should be paid to these two areas in future.
As the progress of the human genome project advances, so does a
clearer understanding of tumor biology, and mo re and more new
prognostic markers with high sensitivity and specificity will be
found and used in clinical assays. However, the combination of some
items, i.e., the pathological features and some biomarkers mentioned
above, seems to be mor e practical for now.
Qin LX, Tang ZY. The prognostic molecular markers in hepatocellular
carcinoma.World J Gastroenterol 2002;8(3):385-392
INTRODUCTION
Liver cancer is one of the common malignancies worldwide, and
has been ranked th e 2nd cancer killer in China since the 1990s.
Although many advances in the clin ical study of hepatocellular
carcinoma (HCC) have been made, and long-term surv ival of patients
has been obtained in some clinical centers, only a definitive s
ubset of cases is cured by surgery, and the overall dismal outcome
of patients w ith HCC has not been completely changed. Lack of
control of metastatic foci and recurrence are the most prevalent
causes of death in patients with HCC, and it is important for tumor
control to identify the factors that predispose patients to death.
Much effort has been made to predict HCC behavior, but specific
prognos tic indicators are still lacking[1].
Prognostic
factors in HCC conventionally consist of staging with the tumor node
metastasis system (TNM) and grading by tumor cellular
differentiation. There are also other factors useful in prognostic
predication but most of them are clinic al. With new discoveries in
cancer biology, pathological and biological factors of HCC in
relation to prognosis have been studied quite extensively.
Morphologic al features of the tumor, both gross and histological,
have been found to signif icantly associate with tumor recurrence
and patient survival[2-4]. A co mplementary way is to
analyze molecular markers for their prognostic significance with
reference to tumor recurrence and survival term in HCC. A large
number of molecular biological factors have been shown to associate
with the invasiveness of HCC, and have potential prognostic
significance. However, routine biomarkers for the prediction of HCC
prognosis are not yet available. In this review, we will focus on
the recent advances in this aspect.
Cellular
malignancy is a very important aspect for patient prognosis. In
recent years, with the development of cellular and molecular
biological techniques, man y molecular markers related to invasion,
metastasis, recurrence and survival hav e been explored. In HCC, DNA
ploidy, the proliferating activity of tumor cells, tumor suppressor
and promoter genes, cell cycle controllers, proteinases that d
egrade extracellular matrix, adhesion molecules, angiogenic factors,
and metabol ic genes, have been regarded as biomarkers for the
malignant phenotype of HCC, and are related to prognosis and
therapeutic outcomes[5].
CELLUAR MALIGNANCY-RELATED MARKERS
DNA-ploidy
Controversy still exists regarding the prognostic
significance of DNA ploidy in HCC patients. Many reports indicate
that DNA ploidy could be a predictive marker for HCC prognosis[6,7].
The overall survival rate of patients with aneuploid cells is much
lower than that of patients with diploid ones, and those wit h
multiple G0/G1 peaks have the worst prognosis. Patients with higher
cell proportions in proliferating stages have a higher early
recurrence rate[7]. However, other studies could not find
a relationship between DNA ploidy and prognosis.
Proliferating activity of HCC cells
Many antigens, such as proliferating cell nuclear antigen (PCNA),
Ki-67, Mcm2, MIB1, MIA, and CSE1L/CAS protein (CAS), have been used
as proliferation markers for cancer cells. The detection of PCNA
with immunohistochemical methods is a co mmon way to study the
proliferating activity of cancer cells[8]. Combine d with
histopathological characteristics, the PCNA labeling index (PCNA-LI)
is one useful marker for evaluating malignant grade, and for
predicting recurrence time and the patients' prognosis of HCC[9].
The
expression of the human Ki-67 protein is strictly associated with
cell prol iferation. The fact that the Ki-67 protein is present
during all active phases of the cell cycle, but is absent from
resting cells, makes it an excellent marke r for determining the
so-called growth fraction of a given cell population. Ki -67 protein
expression is an absolute requirement for progression through the c
ell-division cycle. The fraction of Ki-67-positive tumor cells (the
Ki-67 la beling index) is often correlated with the clinical course
of the disease[1 0,11]. Higher Ki-67 labeling index
(Ki-67-LI) has a very similar clinical significance to PCNA-LI,
reflecting the existence of biologically aggressive ph enotypes and
poor overall and disease-free survival rates in HCC. This could be a
useful factor for predicting the long-term survival of patients with
HCC fol lowing hepatic resection[12,13].
The
CSE1L/CAS protein (CAS) is a Ran-binding protein with a function as
a nuclear transport (export) factor. This protein plays a role in
the mitotic spindle checkpoint, which assures genomic stability
during cell division. This checkpoint is frequently disturbed in
neoplasias of various origins, including hepatic tum ors. The degree
of CAS expression correlates with the grade of tumor dedifferent
iation, and could be a prognostic marker for HCC[14].
Nuclear morphology
Nuclear profiles have been reported as useful prognostic
predictors in various c ancers, including HCC. The nuclear area of
HCC correlates with cell differentiation and cell proliferating
activity, and HCC with a large nuclear area has high potential for
blood vessel invasion and intrahepatic metastasis. Computerized nu
clear morphometry is more objective and quicker than conventional
microscopic an alysis[15]. Recently, quantitative nuclear
morphometry of cancer cells followed by computer-assisted image
analysis (termed Quantitative nuclear grade, QNG) has proven to have
potential use in cancer detection and predicting outcomes such as
tumor stage, recurrence, and progression[16].
p53 gene and its related molecule MDM2
P53 protein plays a central role in cellular responses,
including cell-cycle ar rest and cell death in response to DNA
damage. p53 dysfunction can induce abnorm al cell growth, increased
cell survival, genetic instability, and drug resistanc e. Mutations
in the p53 gene are the most frequently reported somatic gene alter
ation in human cancer. Associations of p53 mutation or positive
immunohistochemi stry staining with higher grade and more advanced
stage has been noted for cance rs of various origins. In addition,
p53 mutation is considered as a strong marke r predicting an
increased risk of local relapse, treatment failure, and overall and
disease-free survival in many kinds of human carcinomas, such as
breast[17-19], colorectal[20], esophageal[21],
head and neck[22], lung[23,24], and ovarian[25],
as well as sarcoma[ 26]. An increased intracellular
concentration of the P53 protein, although no t identical to, is
sometimes seen in tumors with p53 mutation, and has been corr elated
with poor prognosis in some tumor types. Several studies have shown
a rel ationship between the nuclear accumulation of p53 protein and
poor disease-free and overall survival of cancer patients[27,28].
The presence of serum a nti-p53 antibody has also been shown to
associate with survival of patients wit h breast, ovarian, and
colorectal cancer[29,30]. p53 mutations in plasma DNA
could also be detected in cancer patients, and may be used as a
prognostic factor and an early marker to indicate recurrence or
distant metastasis[31 ]. However, there is still a great
controversy as to whether alteration of th e p53 gene adversely
affects survival of cancer patients. Many reports failed to show the
independent prognostic value of p53 in the carcinomas of tongue[32],
breast[33,34], stomach[35], lung[36],
ovarian[37] , bladder[38], colorectal[39],
and non-Hodgkin's lympho ma[40].
In
a similar situation, there are many very controversial results with
the progn ostic value of p53 overexpression or p53 gene mutation in
HCC patients. Many stu dies showed that p53 mutation was involved in
determining the dedifferentiation, the proliferating activity, and
tumor progression[41], was strongly rel ated to the
invasiveness of HCC, and may also influence the postoperative course
(particularly the recurrence within 1 year)[42,43].
Mutations in the p5 3 gene or positive immunostaining for mutant P53
protein expression could be use d as a significant indicator of poor
prognosis. HCCs with p53 mutations have a h igh malignant potential,
and p53 mutation in the primary lesion is useful as an indicator for
the biological behavior of recurrent HCCs. It is also a useful ind
ependent prognostic factor affecting survival after recurrence[9,44,45].
In
a recent prospective study, we found the 3-year and 5-year overall
surviva l rates of HCC patients with positive P53 nuclear
accumulation were much lower than those of the HCC patients with
negative P53 expression. In univariate and mu ltivariate Cox
analysis, p53 overexpression was the most significant factor that
associated with the overall survival rates of HCC patients after
resection. Its significance was even greater than that of factors
such as tumor size, vascular invasion, and tumor capsule, though
they were also related to the overall survi val. p53 mutation or
nuclear accumulation of p53 expression could be a valuable marker
for predicting the prognosis of HCC patients after resection[46]
.
Serum
anti-p53 antibody also could be a useful prognostic factor for HCC
patien ts[47]. However, many different results showed
that neither the immunohi stochemical detection of p53 expression,
nor the serum anti-p53 antibodies had a significant prognostic value
for outcome of patients with HCC[48,49].
The
transcription of the mdm2 gene is activated by p53 and this limits
the growt h-suppressing activity of p53 by direct binding. It has
been reported that MDM2 protein is overexpressed in several types of
cancers. Endo found MDM2 overexpre ssion correlated positively with
p53 mutation, and is a useful predictor of poor prognosis in
patients with HCC following hepatic resection[50].
Cell cycle regulators
Disruption of the G1/S and G2/M check points leads to
uncontrolled cell growth, resulting in the development and
progression of cancers. Overexpression of cycli n A, cyclin D, and
cyclin E have been found to correlate with the tumor relapse of
human HCC, and are independent predictive markers for their
recurrence and pr ognosis[51,52]. The enhanced expression
of cyclin E correlates with hype rphosphorylation of pRb and a high
frequency of Ki-67-positive cells. HCCs wit h enhanced cyclin E
expression probably contain a relatively large number of pro
liferating cancer cells[52]. cdc2 overexpression seems to
play the most crucial role of the modulators in cell cycle
progression and cell proliferation of HCC, and significantly
predicts recurrence[53].
The
p27 protein binds and inhibits cyclin/cyclin-dependent kinase
complexes, is a negative regulator of cell-cycle progression. The
central role of p27 makes it important in a variety of disease
processes, particularly in neoplasia, that involve aberrations in
cellular proliferation and other cell fates. Loss of p27 cooperates
with mutations in several oncogenes and tumor suppressor genes to
fac ilitate tumor growth, indicating that p27 may be a “nodal
point” for tumor su ppression. In most tumor types, reduced p27
expression correlates with poor prog nosis, making p27 a novel and
powerful prognostic marker[54]. High p27 e xpression,
correlated with prolonged survival, is a favorable independent
progno stic parameter for HCC[55,56].
The
protein p73, the first identified homologue of p53 gene, has been
shown to i nduce apoptosis. P73 expression status is significantly
related to prognosis of HCC patients, and could serve as a useful
indicator of prognosis in HCC patients [57]. There is
still controversy with the prognostic value of the p16IN K4a and
p15INK4B genes[58].
Tumor promoter genes and their receptors
Aberrations of many tumor promoter genes, such as ras, c-myc,
c-fms have been indicated as indicators of malignant potential and
poor prognosis i n HCC[9,59-61]. c-myc amplification and
p53 alteration may be copa rticipating events in the progression of
HCC. Disease-free survival in patients showing c-myc amplification
is significantly shorter than in those without
amplification.Hepatocyte growth factor/scatter factor (HGF/SF) is
one of the most important hu moral mediators of liver regeneration.
It is potentially related to molecular me chanisms of
hepatocarcinogenesis via a paracrine system involving its cellular r
eceptor, c-met. Up-regulation of c-met plays an important role in
the development and progression of HCC, and may be a prognostic
marker. Its express ion level is inversely correlated with survival
coordinated with uPA expression [62,63]. However, there
is no significant correlation between the HGF le vel in tumor and
the survival rate of HCC patients[5].
Among of the erb-B receptor family members, c-erbB-2 (Her-2/neu)
represents a well-established prognostic marker and therapeutic
target in several human tu mor types, especially breast cancer.
However, c-erbB-2 is neither a prognostic marker nor a relevant
therapeutic target in human HCCs[9,64]. EGF-R an d
c-erbB-3 play important roles in the progression of HCC, affecting
disease-free survival of HCC patients[5,65].
ets-1
has also been shown to link to cancer invasion and metastasis. ets-1
exp ression was observed with high incidence. However, the average
labeling index (L I) in HCC is lower than in noncancerous lesions.
Even lower expression levels we re found in HCCs of high TNM stage,
poor differentiation, portal invasion, intra hepatic metastasis,
large tumor size, and high Ki-67-LI. HCC patients with hig h ets-1
expression showed better outcomes for disease-free survival than
those with low ets-1 expression[66].
Apoptosis related
The expression of Fas and Fas ligand (Fas L) play a role in
apoptosis of cancer cells including HCCs, and associates with the
prognosis of cancer patients. Fas expression level is significantly
decreased in poorly differentiated HCC and of large size, while Fas
L expression in carcinoma cells is observed exclusively in
moderately or poorly differentiated cases. Each of them has
prognostic signific ance for disease-free survival (DFS)[5,67,68].
Telomerase activity
The ribonucleoprotein telomerase extends telomeres in cancer
cells and has been proposed as a prognostic marker for cancer.
Telomerase activity can be identified as an independent predictor
for recurrence after resection of HCC[69]. The peripheral
blood telomerase activity can also be used as a molecular marker for
the detection of circulating hepatoma cells in blood of HCC
patients, which reflect haematogenous micrometastasis. This is
potentially a practical diagnostic/predictive marker of HCC[70].
Quantitative analysis of telomerase act ivity shows that the
patients with positive telomerase activity in noncancerous liver
tissue have a higher recurrence rate after HCC resection. The
relative tel omerase activity (RTA) of early recurrent patients is
significantly higher than those without recurrence. So, RTA could be
a predictive marker for early recurre nce
after HCC resection[71].
CELL ADHESION AND EXTRACELLULAR MATRIX RELATED
Adhesion molecules
The
expression level of E-cadherin inversely correlates with HCC
histological g rade and prognosis. E-cadherin underexpression might
have some contribution to the early recurrence of HCC[72,73].
In contrast, alpha-, beta-, and ga mma-catenin expression
significantly correlated positively with HCC grade, bein g the
highest in poorly differentiated HCC. Significant positive
associations we re found between gamma-catenin high expression and
capsular invasion or presenc e of satellite nodules, and between
beta-catenin high expression and vascular i nvasion. HCC patients
with underexpression of E-cadherin, alpha-catenin, and g
amma-catenin, and patients with overexpression of beta-catenin, had
poorer sur vival rates[73]. HCCs with a nonnuclear type
of beta-catenin overexpres sion were frequently larger than 5cm in
diameter and had poorer cellular diff erentiation, more
invasiveness, and the patients had significantly shorter disea
se-free survival lengths[74,75]. beta-catenin mutation
associat es with nuclear expression of the protein, and is a
favorable prognostic factor related to low stage[76].
Serum
concentration of intercellular adhesion molecule-1 (sICAM-1) in
patients with HCC is a marker for disease progression and prognosis.
Higher sICAM-1 lev els are more frequently observed in those
patients with multiple lesions and int rahepatic metastasis, and
their prognosis is also very poor. Detecting sICAM-1 is of important
value in predicting tumor recurrence after surgery[77-79]
. The CD44 proteins form a ubiquitously expressed family of cell
surface adhesion molecules involved in cell-cell and cell-matrix
interactions. The major physio logical role of CD44 is to maintain
organ and tissue structure via cell-cell an d cell-matrix adhesion,
but certain variant isoforms can also mediate lymphocyt e activation
and homing, and the presentation of chemical factors and hormones.
The expression of multiple CD44 isoforms is greatly upregulated in
neoplasia. CD 44, particularly its variants, may be useful as a
diagnostic or prognostic marke r of malignancy in at least some
human cancers[80]. Up-regulation of CD 44 isoforms such
as CD44s, CD44v5, CD44v6, CD44v7-8, and CD44v10, correlates wi th
high histological grade, being the highest in poorly differentiated
HCC. CD44 s positivity was an independent factor. Positivity for one
or more CD44 isoforms was the most useful independent factor for
overall survival[81].
Degradation of extracellular matrix
The matrix metalloproteinases (MMP) and the plasminogen
activation system (PA) p lay crucial roles in the process of cancer
invasion and metastasis. Their expres sion levels were found
correlated to recurrence and survival after HCC resection [82,83].
MMP-2, MMP-9, and tissue inhibitors of metalloproteinases -1, -2
(TIMP-1, T IMP-2) have been found to be of prognostic significance
in HCC. The content of MMP-2, MMP-9 in HCC being higher than that in
surrounding liver parenchyma cou ld be used as an important index to
judge the invasion and metastasis of HCC [84,85]. Plasma
MMP-9 levels can also be a candidate for a novel marker for HCC. The
levels appear to reflect the potential and ongoing activity of
vascula r invasion[5].
In
several tumor types, elevated levels of urokinase plasminogen
activator (uPA) , its receptor (uPAR) or its inhibitor plasminogen
activator inhibitor-1 (PAI-1) is associated with a poorer prognosis[85].
uPA activity may be the mo st sensitive factor affecting HCC
invasion in the plasminogen activation system and is a strong
predictor for the recurrence and prognosis of HCC[86,87]
. The PAI-1 proteinis a multifaceted proteolytic factor. It not only
functions as an inhibitor of the protease uPA, but also plays an
important role in signal transduction, cell adherence, and cell
migration. Thus,an apparent paradox cons idering its name-although
it inhibits uPA during blood coagulation, it actually promotes
invasion and metastasis. In many malignancies including HCC,
elevated PAI-1 is associated with tumor aggressiveness and poor
patient outcome[86 ].
ANGIOGENESIS RELATED
Tumor angiogenesis is critical to both the growth and metastasis
of cancer, and is regulated by angiogenic factors. Circulating
angiogenesis regulators have bee n evaluated not only as diagnostic
and/or prognostic factors but also as predict ive factors in cancer
patients. They could be used to determine the risk of deve loping
cancer, to screen for early detection, to distinguish benign from
malignant disease, and to distinguish between different types of
malignancies. In estab lished malignances, they can be uses to
determine prognosis, to predict the resp onse to therapy, and to
monitor the clinical course[5,83,88]. HCC is typically a
hypervascular tumor with a rich blood supply. In recent years, many
angiogenesis-related markers, such as vascular endothelial growth
factor (VEGF), b asic fibroblast growth factor (bFGF), platelet-deriwed
endothelial cell growth factor (PD-ECGF), thrombospondin (TSP),
angiogenin, pleiotrophin and endostatin (ES) levels, as well as
inratumor microvessel density (MVD) have been evaluated and found to
relate to HCC prognosis.
Intratumor microvessel density (MVD)
The intratumor MVD is a direct reflection of tumor
angiogenesis. It can be visua lized by immunohistochemical staining
with antibodies to anti-CD34, Factor VIII, and alpha smooth muscle
actin[89]. MVD levels have a close relationshi p with the
tumor capsule status, tumor size (HCC with 2-5cm in diameter has t
he highest MVD level), intrahepatic recurrence and disease-free
survival, and c an be a predictive marker for disease-free survival[90].
In the authors ' institute, three types of intratumor microvessels,
including capillary-like, sinusoid-like, and mixed-type, were found
in HCC. The MVD level was not relate d to tumor size, capsule
status, Edmondson's grade, or alpha-fetoprotein level; was an
independent factor of disease-free survival in small HCC patients;
and was a predictive marker for early recurrence[91].
Vascular endothelial growth factor (VEGF)
A substantial number of studies have demonstrated a strong
association between elevated tumor expression of VEGF and advanced
disease or poor prognosis in vario us cancers. Circulating VEGF
seems to be a reliable surrogate marker of angiogen ic activity and
tumor progression in cancer patients. It may be predictive of tu mor
status and prognosis in patients with different types of cancer, and
may be useful in predicting and monitoring tumor response to
anticancer therapies and i n follow-up!surveillance for tumor
relapse. It may provide new prognostic infor mation that is not
afforded by conventional clinicopathologic prognostic indicat ors[92].
In HCC, a high serum VEGF level significantly correlates with a
bsence of tumor capsule, presence of intrahepatic metastasis,
presence of micros copic venous invasion, and advanced stage, and it
may be useful as a biologic ma rker of tumor invasiveness and a
prognostic factor in HCC[93]. The data of the authors'
institute also shows serum VEGF is a predictor of invasion and m
etastasis of HCC and a potential biomarker of metastatic recurrence
after curati ve resection[94,95].
Platelet-derived endothelial cell growth factor (PD-ECGF)
PD-ECGF may not be a major regulator of angiogenesis of HCC,
but may play an im portant role in hepatocarcinogenesis, cooperating
with hepatitis C virus. PD-EC GF expression associates with the
venous invasion of HCC[96].
GENOMICS AND PROTEOMICS RELATED
Molecular genetic analyses have clarified that accumulation of
genomic changes p rovides important steps in carcinogenesis and have
identified a number of valuab le genetic markers for certain
cancers. The association of these genomic aberrat ions with the
progression and prognosis of cancer has drawn more and more attent
ion. To date, allelic loss of 1p, 1q21-23, 2p21-16.3, 3p24-p25,
8p22, 8p23, 9 p21, 9q, 10, 13q12, 17p13.3, and 22q13 have been
proposed to be related to the s urvival and prognosis of cancer
patients[97-101].
Many
chromosomal aberrations, including gain of 1q, 8q, and 20q, and loss
of 16q , 4q, 17p, 1p, and 8p have been identified in HCC[102].
However, the rel ationship between these recurrent alterations and
the clinical phenotypes and pr ognosis is still unknown. Towards the
end of 1999, we compared the differences o f chromosomal aberrations
between the primary HCC tumors and their matched metas tatic lesions
using a comparative genomic hybridization (CGH) technique, and fou
nd chromosome 8p deletions might contribute to HCC metastasis[103].
This result was further confirmed by comparison between nude mice
models of HCC with different metastatic potentials[104].
In addition, a more accurate loca tion was identified on 8p23.3,
8p11.2[105]. These findings provide new t argets for
exploring new predictive markers for the recurrence and prognosis of
HCC. Recently, Itano et al. used restriction landmark genomic
scanning (RLGS ), a new high-speed screening method for multiple
genomic changes, to detect un known genetic alterations in HCC. They
found the disease-free survival r ate for patients with > or =16
changed RLGS spots was significantly lower than t hat for patients
with fewer changed RLGS spots (<or =15 spots). In multivariate
analysis, the number of changed spots was proven to retain an
independent progno stic value. These results suggest that the number
of changed RLGS spots may be a useful biological marker for
recurrence of HCC[106].
One
important trend in this area that should be paid attention to is the
prognos tic value of circulating DNA in plasma or serum, and its
genetic alterations in cancer patients. Small amounts of DNA
circulate in both healthy and diseased hum an plasma/serum, and
increased concentrations of DNA are present in the plasma o f cancer
patients. Characteristics of tumor DNA have been found in genetic
mater ial extracted from the plasma of cancer patients. These
features include decreas ed strand stability, the presence of
specific oncogene or tumor suppressor gene mutations, microsatellite
alterations, Ig rearrangements and hypermethylation of several
genes. The results obtained in many different cancers have opened a
new research area indicating that plasma DNA might eventually be a
suitable target for the development of noninvasive diagnostic,
prognostic and follow-up tests f or cancer[107]. Blood
testing for circulating tumor genetic markers may provide valuable
prognostic information and guide future therapy[108].
However,
there is still controversy over the prognostic significance[109].
We found loss of heterozygosity (LOH) on chromosome 14q (D14S62 and
D14S51) could be detected in plasma DNA, and could be of prognostic
significance in HCC patients[1].
Proteomics,
regarded as a sister technology to genomics, is one of the
technologies rapidly changing our approach to understanding tumor
biology. By com paring the proteins present in diseased samples with
those present in normal sam ples, it is possible to identify changes
in expression of proteins that potentia lly may be related to tumor
progression, invasion and metastasis, and prognosis. This technique
has now made it possible to analyze proteins using high th
roughput,automated techniques.Proteomic profiling can be applied to
tissue sam ples as well as body fluids (e.g.serum,urine,etc.),and it
can provide surrogate markers of disease processes,potential
response to treatment,possibility of recu rrence and metastasis for
cancers including HCC[110].
OTHERS
In addition, higher levels of urinary TGF-beta 1[111],
heat shock prote in-27 (HSP-27)[112] and Glutamine
synthetase (GS) expression in the tu mor[113], increased
levels of cyclooxygenase-2 (COX-2) in nontumor liv er tissue[114],
preoperative serum IL-10[115], and HFE mutation s[116]
or down-regulation of DRH1[117] are also powerful progno
stic indicators for shorter disease-free survival and poor
prognosis, related t o tumor progression of HCC.
The
RECK (reversion-inducing-cysteine-rich protein with Kazal motifs)
gene suppresses the invasive and metastatic activities of cancers,
has negative effects on the invasiveness of HCC, and can be regarded
as a promising prognostic molec ular marker for HCC[118].
EXPERIENCES OF THE AUTHORS' INSTITUTE
At the authors' institute, many molecular factors have been
investigated and fou nd to be related to HCC invasiveness in recent
years. They could be divided into two groups: one is positive
invasiveness-related factors, including p16 and p53 mutations, H-ras,
c-erbB2, mdm2, TGF-a, epidermal growth factor receptor (E GFR),
MMP-2, uPA, uPA-R and PAI-1, ICAM-1, VEGF, PD-ECGF, bFGF, and
osteopo ntin (OPN), etc. The other group is negative
invasiveness-related factors, incl uding nm23-H1, Kai-1, TIMP-2,
integrin a5, E-cadherin, etc. These factors co uld be potential
predictive markers for the prognosis of HCC. Serum ICAM-1 and PAI-1
levels were higher in patients with metastasis than those without
metast asis, while serum Thrombomodulin concentration negatively
associated with the in trahepatic spreading and portal vein thrombi
of HCC. Deletions of chromosome 8p and 17p, overexpression of MMP-2,
TGF-a, and EGFR in HCC tissues, and LOH on c hromosome 14q (D14S62
and D14S51) in plasma DNA were also related to metastatic recurrence
and prognosis of HCC patients. p53 mutation or nuclear accumulation
o f p53 expression could be a valuable marker for predicting the
prognosis of HCC patients after resection. E-cadherin, nm23, TIMP-2
are promising prognostic ma rkers[1,
43,46,77,84,86,91,94-96,104-106].
To
search for metastasis-associated genes on a global genomic scale, we
recentl y used cDNA microarrays containing approximately 9984 human
transcripts to inves tigate the gene expression profiles of primary
tumors and their corresponding metastatic lesions (intrahepatic
metastasis or tumor thrombosis of portal vein). A total of 79
significantly upregulated and 69 downregulated genes were identifie
d. Some of them have proven to promote HCC metastasis[119].
These will p rovide new prognostic markers for predicting the
possibillity of metastatic recu rrence and survival after operation.
QUESTIONS AND PROSPECTS
In summary, pathologic factors indicative of tumor invasiveness
such as tumor size, number, capsule state, venous invasion, presence
of satellite nodules, and advanced pTNM stage, are the
best-established risk factors for recurrence and im portant aspects
affecting the prognosis of patients with HCC. Recent molecular r
esearch has identified many tumor biological factors as potential
prognostic mar kers (biomarkers). However, to date, none of them has
been proved to be specific enough, and most of the studies for
specific molecular parameters were correlat ive and retrospective.
Methodologies, sample sizes, and definitions differ. Cons ideration
should be given to the design of prospective clinical trials in
evalua ting the prognostic significance of these markers. These
biomarkers could be detected both in tissue and body fluids (serum,
urine, bile, etc.). Body fluid (particularly blood and urinary)
testing is easily acce ssible and useful in clinical patients, and
is more important in “predicting” the possibility or “early
diagnosis” of recurrence and metastasis. So, future work should be
focused on serum or urinary markers. The prognostic significance of
circulating DNA in plasma or sesum, and its genet ic alterations in
HCC, are important trends that deserve attention. Proteomics and
cDNA array provide other ways to explore new prognostic markers. So,
we can b elieve, with the continuing progress of human genome
project, the development of new molecular and cytogenetic
techniques, and a more complete understanding of tumor biology, more
and more new prognostic markers with high sensitivity and
specificity will soon be found and used in clinical assays. However,
the combination of some items, i.e., pathological features and some
biomarkers mention ed above, seems to be more practical now.
REFERENCES
1
Tang ZY.
Hepatocellular carcinoma-Cause, treatment and metastasis. World J
Gastroenterol 2001;7:445-454
2
Qin LX,
Tang ZY. The prognostic significance of clinical and pathological
features in hepatocellular carcinoma.
World
J Gastroenterol 2002;8:193-199
3
Zhao WH, Ma
ZM, Zhou XR, Feng YZ, Fang BS. Prediction of recurrence and
prognosis in patients with hepatocellular
carcinoma
after resection by use of CL IP score. World J Gastroenterol
2002;8:237-242
4
Zheng N, Ye
SL, Sun RX, Zhao Y, Tang ZY. Effects of cryopreservation and
phenylacetate on biological characters of
adherent
LAK cells from patients with hepatocellular carcinoma. World J
Gastroenterol 2002;8:233-236
5
Korn WM.
Moving toward an understanding of the metastatic process in hep
atocellular carcinoma.
World
J Gastroenterol 2001;7:777-778
6 Mise K,
Tashiro S, Yogita S, Wada D, Harada M, Fukuda Y, Miyake H, Isika wa
M, Izumi K, Sano N. Assessment of the
biological
malignancy of hepatocellular carcinoma: relationship to
clinicopathological factors and prognosis. Clin
Cancer
Res 1998;4:1475-1482
7 Nolte M,
Werner M, Nasarek A, Bektas H, von Wasielewski R, Klempnauer J,
Georgii A. Expression of proliferation
associated
antigens and detection of num erical chromosome aberrations in
primary human liver tumours: relevance to
tumou
r characteristics and prognosis. J Clin Pathol 1998;51:47-51
8 Weber JC,
Nakano H, Bachellier P, Oussoultzoglou E, Inoue K, Shimura H, Wolf
P, Chenard-Neu MP, Jaeck D. Is a
proliferation
index of cancer cells a rel iable prognostic factor after
hepatectomy in patients with colorectal liver meta
stases?
Am J Surg 2001;182:81-88
9 Lin GY,
Chen ZL, Lu CM, Li Y, Ping XJ, Huang R. Immunohistochemical stud y
on p53, H-rasp21, c-erbB-2 protein and PCNA
expression
in HCC tissues of Ha n and minority ethnic patients. World J
Gastroenterol 2000;6:234-238
10 Scholzen T, Gerdes
J. The Ki-67 protein: from the known and t he unknown. J Cell
Physiol 2000;182:311-322
11 Hernandez-Rodriguez
NA, Correa E, Sotelo R, Contreras-Paredes A, Gom ez-Ruiz C, Green L,
Mohar A. Ki-67: a
proliferative
marker that may predict pu lmonary metastases and mortality of
primary osteosarcoma. Cancer Detect
Prev
2001;25:210-215
12 Ito Y, Matsuura N,
Sakon M, Takeda T, Umeshita K, Nagano H, Nakamori S , Dono K,
Tsujimoto M, Nakahara M, Nakao K,
Monden
M. Both cell proliferation an d apoptosis significantly predict
shortened disease-free survival in hepatocell ular
carcinoma.
Br J Cancer 1999;81:747-751
13 Ouchi K, Sugawara T,
Ono H, Fujiya T, Kamiyama Y, Kakugawa Y, Mikuni J , Yamanami H,
Komatsu S, Horikoshi A.
Mitotic
index is the best predictive facto r for survival of patients with
resected hepatocellular carcinoma. Dig Surg
2000;17:42-48
14 Wellmann A, Flemming
P, Behrens P, Wuppermann K, Lang H, Oldhafer K, P astan I, Brinkmann
U. High expression of
the
proliferation and apoptosis associa ted CSE1L/CAS gene in hepatitis
and liver neoplasms: correlation with tumor
prog
ression. Int J Mol Med 2001;7:489-494
15 Ikeguchi M, Sato N,
Hirooka Y, Kaibara N. Computerized nuclear morphom etry of
hepatocellular carcinoma and its
relation
to proliferative activity. J Surg Oncol 1998;68:225-230
16 Veltri RW, Partin
AW, Miller MC. Quantitative nuclear grade (QNG): a n ew image
analysis-based biomarker of clinically
relevant
nuclear structure alte rations. J Cell Biochem 2000;Suppl 35:151-157
17 Overgaard J, Yilmaz
M, Guldberg P, Hansen LL, Alsner J. TP53 mutation is an independent
prognostic marker for poor
outcome
in both node-negative and node-positive breast cancer. Acta Oncol
2000;39:327-333
18 Takahashi M, Tonoki
H, Tada M, Kashiwazaki H, Furuuchi K, Hamada J, Fu jioka Y, Sato Y,
Takahashi H, Todo S,
Sakuragi
N, Moriuchi T. Distinct prognosti c values of p53 mutations and loss
of estrogen receptor and their cumulative
eff
ect in primary breast cancers. Int J Cancer 2000;89:92-99
19 Blaszyk H, Hartmann
A, Cunningham JM, Schaid D, Wold LE, Kovach JS, So mmer SS. A
prospective trial of midwest
breast
cancer patients: a p53 gene mutat ion is the most important
predictor of adverse outcome. Int J Cancer 2000;
89
:32-38
20 Kahlenberg MS,
Stoler DL, Rodriguez-Bigas MA, Weber TK, Driscoll DL, Anderson GR,
Petrelli NJ. p53 tumor suppressor
gene
mutations predict decreased survival of patients with sporadic
colorectal carcinoma. Cancer 2000;88:1814 -1819
21 Ireland AP, Shibata
DK, Chandrasoma P, Lord RV, Peters JH, DeMeester T R. Clinical
significance of p53 mutations in
adenocarcinoma
of the esophagus and cardia. Ann Surg 2000;231:179-187
22 Tamas L, Kraxner H,
Mechtler L, Repassy G, Ribari O, Hirschberg A, Szentkuti G, Jaray B,
Szentirmay Z. Prognostic
significance
of P53 histochem istry and DNA histogram parameters in head and neck
malignancies. Anticancer R es
2000;20:4031-4037
23 Murakami I, Hiyama
K, Ishioka S, Yamakido M, Kasagi F, Yokosaki Y. p53 gene mutations
are associated with shortened
survival
in patients with advanced non-small cell lung cancer: an analysis of
medically managed patients. Clin Cancer
Res
2000;6:526-530
24 Mitsudomi T,
Hamajima N, Ogawa M, Takahashi T. Prognostic significance of p53
alterations in patients with non-small
cell
lung cancer: a meta-analys is. Clin Cancer Res 2000;6:4055-4063
25 Shahin MS, Hughes JH,
Sood AK, Buller RE. The prognostic significance of p53 tumor
suppressor gene alterations in
ovarian
carcinoma. Cancer 2000;8 9:2006-2017
26 de Alava E,
Antonescu CR, Panizo A, Leung D, Meyers PA, Huvos A G, Pardo-Mindan
FJ, Healey JH, Ladanyi M.
Prognostic
impact of P53 status in Ew ing sarcoma. Cancer 2000;89:783-792
27 Leibovich BC, Cheng
L, Weaver AL, Myers RP, Bostwick DG. Outcome predi ction with p53
immunostaining after radical
prostatectomy
in patients with local ly advanced prostate cancer. J Urol
2000;163:1756-1760
28 Osaki T, Kimura T,
Tatemoto Y, Dapeng L, Yoneda K, Yamamoto T. Diffuse mode of tumor
cell invasion and expression of
mutant
p53 protein but not of p21 protein are correlated with treatment
failure in oral carcinomas and their meta static
foci. Oncology
2000;59:36-43
29 Suzuki M, Ohwada M,
Saga Y, Kohno T, Takei Y, Sato I. Micrometastatic p53-positive cells
in the lymph nodes of early
stage
epithelial ovarian cancer: prognostic significance. Oncology
2001;60:170-175
30 Shiota G, Ishida M,
Noguchi N, Oyama K, Takano Y, Okubo M, Katayama S, Tomie Y, Harada
K, Hori K, Ashida K,
Kishimoto
Y, Hosoda A, Suou T, Kanbe T, Ta naka K, Nosaka K, Tanida O, Kojo H,
Miura K, Ito H, Kaibara N, Kawasaki H.
Circulating
p53 antibody in patients with colorectal cancer: relation to
clinicopatho logic features and survival. Dig Dis
Sci
2000;45:122-128
31 Shao ZM, Wu J, Shen
ZZ, Nguyen M. p53 mutation in plasma DNA and its p rognostic value
in breast cancer patients.
Clin
Cancer Res 2001;7:2222-2227
32 Kantola S, Parikka
M, Jokinen K, Hyrynkangs K, Soini Y, Alho OP, Salo T. Prognostic
factors in tongue cancer - relsative
importance
of demographic, c linical and histopathological factors. Br J Cancer
2000;83:614-619
33 Ferrero JM, Ramaioli
A, Formento JL, Francoual M, Etienne MC, Peyrotte s I, Ettore F,
Leblanc-Talent P, Namer M,
Milano
G. P53 determination alongside classical prognostic factors in
node-negative breast cancer: an evaluation at
more
than 10-year follow-up. Ann Oncol 2000;11:393-397
34 Reed W, Hannisdal E,
Boehler PJ, Gundersen S, Host H, Marthin J . The prognostic value of
p53 and c-erb B-2
immunostaining
is overrated for pa tients with lymph node negative breast
carcinoma: a multivariate analysis of pro
gnostic
factors in 613 patients with a follow-up of 14-30 years. Cancer 20
00;88:804-813
35 Kaye PV, Radebold K,
Isaacs S, Dent DM. Expression of p53 and p21waf1/ cip1 in gastric
carcinoma: lack of
inter-relationship
or correlation with progn osis. Eur J Surg Oncol 2000;26:39-43
36 Schiller JH, Adak S,
Feins RH, Keller SM, Fry WA, Livingston RB, Hammo nd ME, Wolf B,
Sabatini L, Jett J, Kohman L,
Johnson
DH. Lack of prognostic sign ificance of p53 and K-ras mutations in
primary resected non-small-cell lung c ancer
on
E4592: a Laboratory Ancillary Study on an Eastern Cooperative
Oncology Group Prospective Randomized Trial of
Postoperative
Adjuvant Therapy. J Clin O ncol 2001;19:448-457
37 Gadducci A, Cianci
C, Cosio S, Carnino F, Fanucchi A, Buttitta F, Cont e PF, Genazzani
AR. p53 status is neither a
predictive
nor a prognostic variable in patients with advanced ovarian cancer
treated with a paclitaxel-based regim en.
Anticancer
Res 2000;20:4793-4799
38 Fleshner N, Kapusta
L, Ezer D, Herschorn S, Klotz L. p53 nuclear accum ulation is not
associated with decreased
disease-free
survival in patients with node positive transitional cell carcinoma
of the bladder. J Urol 2000;164:
1177-1182
39 Gallego MG, Acenero
MJ, Ortega S, Delgado AA, Cantero JL. Prognostic i nfluence of p53
nuclear overexpression in
colorectal
carcinoma. Colon Rectum 2000;43:971-975
40 Nieder C, Petersen
S, Petersen C, Thames HD. The challenge of p53 as p rognostic and
predictive factor in Hodgkin's or
non-Hodgkin's
lymphoma. Ann H ematol 2001;80:2-8
41 Itoh T, Shiro T,
Seki T, Nakagawa T, Wakabayashi M, Inoue K, Okamura A . Relationship
between p53 overexpression
and
the proliferative activity in hepa tocellular carcinoma. Int J Mol
Med 2000;6:137-142
42 Jeng KS, Sheen IS,
Chen BF, Wu JY. Is the p53 gene mutation of prognos tic value in
hepatocellular carcinoma after
resection?
Arch Surg 2000;135:13 29-1333
43 Tang ZY, Qin LX,
Wang XM, Zhou G, Liao Y, Weng Y, Jiang XP, Lin ZY, Li u KD, Ye SL.
Alterations of oncogenes, tumor
suppressor
genes and growth factors in hepatocellular carcinoma: with relation
to tumor size and invasiveness.
Chin
Med J 1998;111:313-318
44 Sugo H, Takamori S,
Kojima K, Beppu T, Futagawa S. The significance of p53 mutations as
an indicator of the biological
behavior
of recurrent hepatocel lular carcinomas. Surg Today 1999;29:849-855
45 Heinze T, Jonas S,
Karsten A, Neuhaus P. Determination of the oncogene s p53 and C-erb
B2 in the tumour cytosols of
advanced
hepatocellular carcinoma (HCC) and correlation to survival time.
Anticancer Res 1999;19:2501-2503
46 Qin LX, Tang ZY, Ma
ZC, Wu ZQ, Zhou XD, Ye QH, Ji Y, Huang LW, Jia HL, Sun HC, Wang L.
P53 immunohistochemistry
scoring
is an independent prognostic m arker of patient with hepatocellular
carcinoma resection: A prospective study of
256
formalin-fixed paraffin-embedded tumor samples. World J
Gastroenterol 2002(in press)
47 Shiota G, Kishimoto
Y, Suyama A, Okubo M, Katayama S, Harada K, Ishida M, Hori K, Suou
T, Kawasaki H. Prognostic
significance
of serum anti-p53 antib ody in patients with hepatocellular
carcinoma. J Hepatol 1997;27:661-668
48 Tangkijvanich P,
Janchai A, Charuruks N, Kullavanijaya P, Theamboonler s A, Hirsch P,
Poovorawan Y. Clinical
associations
and prognostic significance o f serum anti-p53 antibodies in Thai
patients with hepatocellular carcinoma.
A
sian Pac J Allergy Immunol 2000;18:237-243
49 Saffroy R, Lelong JC,
Azoulay D, Salvucci M, Reynes M, Bismuth H, Debu ire B, Lemoine A.
Clinical significance of
circulating anti-p53 antibodies in E uropean patients with
hepatocellular carcinoma. Br J Cancer 1999;79:604-610
50 Endo K, Ueda T, Ohta
T, Terada T. Protein expression of MDM2 and its c linicopathological
relationships in human
hepatocellular
carcinoma. Liver 20 00;20:209-215
51 Chao Y, Shih YL,
Chiu JH, Chau GY, Lui WY, Yang WK, Lee SD, Huang TS. Overexpression
of cyclin A but not Skp 2
correlates
with the tumor relapse of hu man hepatocellular carcinoma. Cancer
Res 1998;58:985-990
52 Ohashi R, Gao C,
Miyazaki M, Hamazaki K, Tsuji T, Inoue Y, Uemura T, H irai R,
Shimizu N, Namba M. Enhanced
expression
of cyclin E and cyclin A in huma n hepatocellular carcinomas.
Anticancer Res 2001;21:657-662
53 Ito Y, Takeda T,
Sakon M, Monden M, Tsujimoto M, Matsuura N. Expressio n and
prognostic role of cyclin-dependent
kinase
1 (cdc2) in hepatocellular car cinoma. Oncology 2000;59:68-74
54 Philipp-Staheli J,
Payne SR, Kemp CJ. p27(Kip1): regulation and funct ion of a
haploinsufficient tumor suppressor and its
misregulation
in cancer. E xp Cell Res 2001;264:148-168
55 Fiorentino M,
Altimari A, D'Errico A, Cukor B, Barozzi C, Loda M, Grig ioni WF.
Acquired expression of p27 is a favorable
prognostic
indicator in patie nts with hepatocellular carcinoma. Clin Cancer
Res 2000;6:3966-3972
56 Ito Y, Matsuura N,
Sakon M, Miyoshi E, Noda K, Takeda T, Umeshita K, N agano H,
Nakamori S, Dono K, Tsujimoto M,
Nakahara
M, Nakao K, Taniguchi N, Mond en M. Expression and prognostic roles
of the G1-S modulators in
hepatocellular
carcinoma: p27 independently predicts the recurrence. Hepatology
1999;30:90 -99
57 Tannapfel A, Wasner
M, Krause K, Geissler F, Katalinic A, Hauss J, Mossner J, Engeland
K, Wittekind C. Expression of
p73
and its relation to histopath ology and prognosis in
hepatocellular carcinoma. J Natl Cancer Inst 1999;91: 1154-1158
58
Qin Y, Li B, Tan YS,
Sun ZL, Zuo FQ, Sun ZF. Polymorphism of p16INK4a gene and rare
mutation of p15INK4b gene
exon2
in primary hepatocarcinoma. Worl d J Gastroenterol 2000;6:411-414
59
Wang Q, Lin ZY,
Feng XL. Alterations in metastatic properties of hepat ocellular
carcinoma cell following H-ras oncogene
transfection.
World J Gastr oenterol 2001;7:335-339
60 Cui J, Yang DH, Bi
XJ, Fan ZR. Methylation status of c-fms oncogenein HCC and its
relationship with clinical pathology.
World
J Gastroenterol 200 1;7:136-139
61 Kawate S, Fukusato
T, Ohwada S, Watanuki A, Morishita Y. Amplification of c-myc in
hepatocellular carcinoma:
correlation
with clinicopathologic f eatures, proliferative activity and p53
overexpression. Oncology 1999;57:157 -163
62 Tavian D, De Petro
G, Benetti A, Portolani N, Giulini SM, Barlati S. u -PA and c-MET
mRNA expression is co-ordinately
enhanced
while hepatocyte grow th factor mRNA is down-regulated in human
hepatocellular carcinoma. Int J Can cer
2000;87:644-649
63
Luo YQ, Wu MC, Cong WM.
Gene expression of hepatocyte growth factor an d its receptor in HCC
and nontumorous
liver
tissues. World J Gastroenterol 1 999;5:119-121
64 Prange W,
Schirmacher P. Absence of therapeutically relevant c-erbB- 2
expression in human hepatocellular
carcinomas.
Oncol Rep 2001;8:727-730
65 Ito Y, Takeda T,
Sakon M, Tsujimoto M, Higashiyama S, Noda K, Miyoshi E, Monden M,
Matsuura N. Expression and
clinical
significance of erb-B receptor family in hepatocellular carcinoma.
Br J Cancer 2001;84:1377-1383
66 Ito Y, Miyoshi E,
Takeda T, Sakon M, Noda K, Tsujimoto M, Monden M, Ta niguchi N,
Matsuura N. Expression and
possible
role of ets-1 in hepatocellular carcinoma. Am J Clin Pathol
2000;114:719-725
67 Ito Y, Monden M,
Takeda T, Eguchi H, Umeshita K, Nagano H, Nakamori S, Dono K, Sakon
M, Nakamura M, Tsujimoto
M,
Nakahara M, Nakao K, Yokosaki Y, Mats uura N. The status of Fas and
Fas ligand expression can predict recurrence
of
he patocellular carcinoma. Br J Cancer 2000;82:1211-1217
68 Wang XZ, Chen XC,
Yang YH, Chen ZX, Huang YH, Tao QM. Relationship bet ween HBxAg and
Fas/FasL in patients with
hepatocellular
carcinoma. World J Gas troenterol 2000;6:S17
69 Kobayashi T, Kubota
K, Takayama T, Makuuchi M. Telomerase activity as a predictive
marker for recurrence of
hepatocellular
carcinoma after hepatectomy . Am J Surg 2001;181:284-288
70 Tatsuma T, Goto S,
Kitano S, Lin YC, Lee CM, Chen CL. Telomerase activ ity in
peripheral blood for diagnosis of
hepatoma.
J Gastroenterol Hepatol 2 000;15:1064-1070
71 Suda T, Isokawa O,
Aoyagi Y, Nomoto M, Tsukada K, Shimizu T, Su zuki Y, Naito A,
Igarashi H, Yanagi M, Takahashi T,
Asakura
H. Quantitation of t elomerase activity in hepatocellular carcinoma:
a possible aid for a prediction of recurrent
diseases
in the remnant liver. Hepatology 1998;27:402-406
72 Huang GT, Lee HS,
Chen CH, Sheu JC, Chiou LL, Chen DS. Correlation of E-cadherin
expression and recurrence of
hepatocellular
carcinoma. Hepatogastr oenterology 1999;46:1923-1927
73 Endo K, Ueda T,
Ueyama J, Ohta T, Terada T. Immunoreactive E-cadherin , alpha-catenin,
beta-catenin, and
gamma-catenin
proteins in hepatocellular c arcinoma: relationships with tumor
grade, clinicopathologic parameters,
and
pati ents' survival. Hum Pathol 2000; 31:558-565
74 Wong CM, Fan ST, Ng
IO. beta-Catenin mutation and overexpression in h epatocellular
carcinoma: clinicopathologic
and
prognostic significance. Cancer 2001;92:136-145
75
Cui J, Zhou XD, Liu YK,
Tang ZY, Zile MH. Abnormal Cu-catenin gene ex pression with
invasiveness of primary
hepatocellular
carcinoma in China. World J Gastroenterol 2001;7:542-546
76 Hsu HC, Jeng YM, Mao
TL, Chu JS, Lai PL, Peng SY. Beta-catenin mutati ons are associated
with a subset of low-stage
hepatocellular
carcinoma negative for hepatitis B virus and with favorable
prognosis. Am J Pathol 2000;157:76 3-770
77 Sun JJ, Zhou XD, Liu
YK, Tang ZY, Feng JX, Zhou G. Xue Q, Chen J. Inva sion and
metastasis of liver cancer: expression
of
intercellular adhesion molecu le-1. J Cancer Res Clin Oncol
1999;125:28-34
78
Mei MH, Xu J, Shi QF,
Yang JH, Chen Q, Qin LL. Clinical significance o f serum
intercellular adhesion molecule-1
detection
in patients with hepatocell ular carcinoma. World J Gastroenterol
2000;6:408-410
79
Xu J, Mei MH, Zeng SE,
Shi QF, Liu YM, Qin LL. Expressions of ICAMa21 and its mRNA in sera
and tissues of patients
with
hepatocellular carcinoma. Wo rld J Gastroenterol 2001;7:120-125
80 Goodison S, Urquidi
V, Tarin D. CD44 cell adhesion molecules. Mol Pa thol
1999;52:189-196
81 Endo
K, Terada T. Protein expression of CD44 (standard and variant iso
forms) in hepatocellular carcinoma: relationships
with
tumor grade, clinicopatho logic parameters, p53 expression, and
patient survival. J Hepatol 2000;32:78 -84
82 Sakamoto Y, Mafune
K, Mori M, Shiraishi T, Imamura H, Mori M, Takayama T, Makuuchi M.
Overexpression of MMP-9
correlates
with growth of small hepatoc ellular carcinoma. Int J Oncol
2000;17:237-243
83
Jiang YF, Yang ZH, Hu
JQ. Recurrence or metastasis of HCC: predictors, early detection and
experimental antiangiogenic
therapy.
World J Gastroentero l 2000;6:61-65
84 Bu W, Tang ZY, Ye
SL, Liu KD, Huang XW, Gao DM. The association of type IV collagenase
with invasion and metastasis
of
hepatocellular carcinoma . Zhonghua Xiaohua Zazhi 1999;19:13-15
85 Fox SB, Taylor M,
Grondahl-Hansen J, Kakolyris S, Gatter KC, H arris AL. Plasminogen
activator inhibitor-1 as a measure
of
vascular remodelli ng in breast cancer. J Pathol 2001;195:236-243
86 Zheng Q, Tang ZY,
Xue Q, Shi DR, Song HY, Tang HB. Invasion and metast asis of
hepatocellular carcinoma in relation to
urokinase-type
plasminogen acti vator, its receptor and inhibitor. J Cancer Res
Clin Oncol 2000;126:641-646
87 Itoh T, Hayashi Y,
Kanamaru T, Morita Y, Suzuki S, Wang W, Zhou L, Rui JA, Yamamoto M,
Kuroda Y, Itoh H. Clinical
significance
of urokinase-ty pe plasminogen activator activity in hepatocellular
carcinoma. J Gastroenterol Hepatol
2000;15:422-430
88 Kuroi K, Toi M.
Circulating angiogenesis regulators in cancer patients . Int J Biol
Markers 2001;16:5-26
89 Morinaga S, Imada T,
Shimizu A, Akaike M, Sugimasa Y, Takemiya S, Taka nashi Y.
Angiogenesis in hepatocellular
carcinoma
as evaluated by alpha smooth m uscle actin immuno histochemistry.
Hepatogastroenterology 2001;48:
224-228
90 El-Assal ON, Yamanoi
A, Soda Y, Yamaguchi M, Igarashi M, Yamamoto A, Nabika T, Nagasue N.
Clinical significance of
microvessel
density and vascular e ndothelial growth factor expression in
hepatocellular carcinoma and surrounding
liver:
possible involvement of vascular endothelial growth factor in the
angioge nesis of cirrhotic liver. Hepatology
1998;27:1554-1562
91 Sun HC, Tang ZY, Li
XM, Zhou YN, Sun BR, Ma ZC. Microvessel density of hepatocellular
carcinoma: its relationship with
prognosis.
J Cancer Res Clin Oncol 1999;125:419-26
92 Poon RT, Fan ST,
Wong J. Clinical implications of circulating angiogen ic factors in
cancer patients. J Clin Oncol
2001;19:1207-1225
93 Poon RT, Ng IO, Lau
C, Zhu LX, Yu WC, Lo CM, Fan ST, Wong J. Serum vas cular endothelial
growth factor predicts
venous
invasion in hepatocellular carci noma: a prospective study. Ann Surg
2001;233:227-235
94 Li XM, Tang ZY, Qin
LX, Zhou J, Sun HC. Serum vascular endothelial gro wth factor is a
predictor of invasion and
metastasis
in hepatocellular carcinoma . J Exp Clin Cancer Res 1999,18:511-517
95
Niu Q, Tang ZY, Ma ZC,
Qin LX, Zhang LH. Serum vascular endothelial growth factor is a
potential biomarker of
metastatic
recurrence after curative res ection of hepatocellular carcinoma.
World J Gastroenterol 2000;6: 565-568
96 Zhou J, Tang ZY, Fan
J, Wu ZQ, Li XM, Liu YK, Liu F, Sun HC, Ye SL. Ex pression of
platelet-derived endothelial cell
growth
factor and vascular endoth elial growth factor in hepatocellular
carcinoma and portal vein tumor thrombus.
J
Cancer Res Clin Oncol 2000; 126:57-61
97 Tada K, Shiraishi S,
Kamiryo T, Nakamura H, Hirano H, Kuratsu J, Kochi M, Saya H, Ushio
Y. Analysis of loss of
heterozygosity
on chromosome 10 in pati ents with malignant astrocytic tumors:
correlation with patient age and
survival
. J Neurosurg 2001;95:651-659
98 Bisgaard ML, Jager
AC, Dalgaard P, Sondergaard JO, Rehfeld JF, Nielsen FC. Allelic loss
of chromosome 2p21-16.3 is
associated
with reduced survival i n sporadic colorectal cancer. Scand J
Gastroenterol 2001;36:405-409
99 Hirano A, Emi M,
Tsuneizumi M, Utada Y, Yoshimoto M, Kasumi F, Akiyama F, Sakamoto G,
Haga S, Kajiwara T,
Nakamura
Y. Allelic losses of loci at 3p25.1 , 8p22, 13q12, 17p13.3, and
22q13 correlate with postoperative recurrence in
bre
ast cancer. Clin Cancer Res 2001;7:876-882
100 Simoneau M, LaRue H, Aboulkassim TO, Meyer F, Moore L, Fradet Y.
Chro mosome 9 deletions and recurrence of
superficial
bladder cancer: identification of four regions of prognostic
interest. Oncogene 2000;19:6317-6323
101 Washburn JG, Wojno KJ, Dey J, Powell IJ, Macoska JA. 8pter- p23
dele tion is associated with racial differences in
prostate
cancer outcome. Clin Ca ncer Res 2000;6:4647-4652
102 Wong N, Lai P, Lee SW, Fan S, Pang E, Liew CT, Sheng Z, Lau JW,
Johns on PJ. Assessment of genetic changes in
hepatocellular
carcinoma by comparative genomic hybridization analysis:
relationship to disease stage, tumor size, and c
irrhosis.
Am J Pathol 1999;154: 37-43
103 Itano O, Ueda M, Kikuchi K, Shimazu M, Kitagawa Y, Aiura K,
Kitajima M. A new predictive factor for hepatocellular
carcinoma
based on two-dimensiona l electrophoresis of genomic DNA. Oncogene
2000;19:1676-1683
104 Qin LX, Tang ZY, Sham JST, Ma ZC, Ye SL, Zhou XD. The associat
ion of chromosome 8p deletion and tumor metastasis
in
human hepatocellular carci noma. Cancer Res 1999;59:5662-5665
105 Qin LX, Tang ZY, Ye SL, Liu YK, Ma ZC, Zhou XD, Wu ZQ, Lin ZY,
Sun FX, Tian J, Guan XY, Pack SD, Zhuang ZP.
Chromosome
8p deletion is associat ed with metastasis of human hepatocellular
carcinoma when high and low
metastati
c models are compared. J Cancer Res Clin Oncol 2001;127:482-488
106 Zhang LH, Qin LX, Ma ZC, Ye SL, Liu YK, Ye QH, Wu X, Huang W,
Tang ZY . Identification of allelic imbalances regions
related
to metastasis of hepatoce llular carcinoma: Comparison between
matched primary and metastatic lesions in 2
2
patients by genome-wide microsatellite Analysis. Int J Cancer 2002 (submi
tted)
107 Anker P, Stroun M. Circulating DNA in plasma or serum. Medicina
( B Aires) 2000;60:699-702
108 Taback B, Fujiwara Y, Wang HJ, Foshag LJ, Morton DL, Hoon DS.
Prognos tic significance of circulating microsatellite
markers
in the plasma of melanoma patients. Cancer Res 2001;61:5723-5726
109 Nunes DN, Kowalski LP, Simpson AJ. Circulating tumor-derived DNA
may permit the early diagnosis of head and neck
squamous
cell carcinomas. Int J C ancer 2001;92:214-219
110 Kennedy S. Proteomic profiling from human samples: the body
fluid alt ernative. Toxicol Lett 2001;120:379-384
111 Tsai JF, Chuang LY, Jeng JE, Yang ML, Chang WY, Hsieh MY, Lin ZY,
Tsa i JH. Clinical relevance of transforming growth
factor-
beta 1 in the urine of p atients with hepatocellular carcinoma.
Medicine (Baltimore) 1997;76:213-226
112 King KL, Li AF, Chau GY, Chi CW, Wu CW, Huang CL, Lui WY.
Prognostic significance of heat shock protein-27
expression
in hepatocellular carcinoma an d its relation to histologic grading
and survival. Cancer 2000;88:2464-2470
113 Osada T, Nagashima I, Tsuno NH, Kitayama J, Nagawa H. Prognostic
sign ificance of glutamine synthetase expression
in
unifocal advanced hepatocellular carcinoma. J Hepatol
2000;33:247-253
114 Kondo M, Yamamoto H, Nagano H, Okami J, Ito Y, Shimizu J, Eguchi
H, M iyamoto A, Dono K, Umeshita K, Matsuura N,
Wakasa
K, Nakamori S, Sakon M, Monden M. Increased expression of COX-2 in
nontumor liver tissue is associated withs
horter
disease-free survival in patients with hepatocellular carcinoma.
Clin Cancer Res 1999;5:4005-4012
115 Chau GY, Wu CW, Lui WY, Chang TJ, Kao HL, Wu LH, King KL, Loon g
CC, Hsia CY, Chi CW. Serum interleukin-10 but not
interleukin-6
is related t o clinical outcome in patients with resectable
hepatocellular carcinoma. Ann S urg
2000;231:552-558
116 Pirisi M, Toniutto P, Uzzau A, Fabris C, Avellini C, Scott C,
Apollon io L, Beltrami CA, Bresadola F. Carriage of HFE
mutations
and
outcome of surgica l resection for hepatocellular carcinoma
in cirrhotic patients. Cancer 2000; 89:297-302
117 Yamamoto Y, Sakamoto M, Fujii G, Kanetaka K, Asaka M, Hirohashi
S. Cl oning and characterization of a novel gene,
DRH1,
down-regulated in advanced hu man hepatocellular carcinoma. Clin
Cancer Res 2001;7:297-303
118 Furumoto K, Arii S, Mori A, Furuyama H, Gorrin Rivas MJ, Nakao
T, Iso be N, Murata T, Takahashi C, Noda M, Imamura
M.
RECK gene expression in hepatoce llular carcinoma: correlation with
invasion-related clinicopathological factors and
its
clinical significance. Reverse-inducing-cysteine-rich protein with
Kazal motifs. Hepatology 2001;33:189-195
119 Ye QH, Qin LX, Forgues M, He P, Kim JW, Peng AC, Simon R, Robles
A, C hen YD, Ma ZC, Wu ZQ, Ye SL, Liu YK, Tang
ZY,
Wang XW. Gene expression profiling and supervised machine learning
to define metastasis-related genes in human
he
patocellular carcinoma. Nature Med 2002 (submitted)
Edited
by Pagliarini
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