close

Вход

Забыли?

вход по аккаунту

?

620

код для вставкиСкачать
1195
Flow Cytometric DNA Analysis of Cirrhotic Liver Cells
in Patients with Hepatocellular Carcinoma Can
Provide a New Prognostic Factor
Savino Rua, M.D.'
Alberto Comino, M . D . ~
Adriana Fruttero, Ph.0.'
Pierfederico Torchio, M.D?
Hedayat Bouzari, M . D . ~
Stefan0 Taraglio, M . D . ~
Bruno Torchio, M.D?
Lorenzo Capussotti, M.D."
' Department of Pathological Anatomy and His-.
tology, City Hospital, Cuneo, Italy.
* Department of Pathological Anatomy and His-.
tology, City Hospital, Mondovi, Italy.
Institute of Statistics, University of Milan, Mi..
lam, Italy.
Department of Surgery, Hospital, Mauriziano
"Umberto I st.", Torino, Italy.
Department of Pathological Anatomy and Histology, Hospital, Mauriziano "Umberto I st.",
Torino, Italy.
BACKGROUND. DNA flow cytometry of hepatocellular carcinoma (HCC) cells has
been investigated in many studies, but, to the best of our knowledge, there are no
data on DNA analysis of cirrhotic parenchyma around the HCC. In this study, cell
kinetics and ploidy of parenchymal cells around HCC were performed to ascertain
if this would predict the possibility of recurrence in the cirrhotic areas.
METHODS. The DNA content of 93 cases of HCC and of cirrhotic liver around the
tumor nodules was analyzed by flow cytometry. Ploidy and proliferative index of
HCC and cirrhotic liver were compared with macroscopic, histologic, and clinical
features of each case and linked with the behavior of these tumors. Survival curves
were assessed according to the Kaplan-Meier method. A multivariate analysis
based on Cox proportional hazards regression model was performed on cases of
diploid cirrhosis cells in which the S-phase fraction was evaluable.
RESULTS. The univariate analysis of survival suggested significant roles for age,
number of intrahepatic nodules, Edmondson-Steiner's classification, portal invasion, vascular invasion, presence of necrosis, hepatitis B surface antigen, u-fetoprotein, Child's score, ploidy, and S-phase fraction of HCC cells. The DNA analysis
of the cirrhotic cells showed that polyploidy was dramatically reduced in patients
with HCC, compared with normal hepatocytes, and aneuploid clones were present
among diploid cells. High S-phase fraction of cirrhotic cells and Child-Pugh classification were the strongest independent parameters affecting the tumor behavior
in this study.
CONCLUSIONS. The results of this study suggest that S-phase fraction of cirrhotic
liver parenchyma may be employed as a new parameter in the prognostic evaluation of HCC patients. Cancer 1996; 78:1195-1202.0 1996Arnericun Cancer Society.
KEYWORDS: DNA flow cytometry, S-phase fraction, hepatocellularcarcinoma, cinhosis.
M
Addiress for reprints: Savino Rua, M.D., Department of Pathological Anatomy and Histology,
City Hospital, Via Coppino 26, 12100 Cuneo,
Italy.
Received November 13, 1995; revisions received February 20, 1996 and May 31, 1996;
accepted May 31, 1996.
0 'I996 American Cancer Society
ost hepatocellular carcinomas (HCC)arise in cirrhotic patients. Hepatocyte proliferation is significantlycorrelated to the development
of HCC in these patients.' In fact, those with a high cell proliferation
run an increased risk of developing HCC and may require a differentiated
follow-up.2Moreover, precancerous lesions, such as hyperplastic or adenomatous nodules, which may be present in cirrhotic livers, could give
rise to tumor recurrence in surgically treated HCC ~atients.3.~
The prognosis of patients with HCC depends both on morphologic
factors, such as tumor size and differentiation,presence of satellite nodules, and vascular invasion, and on clinical factors, such as patient's age,
general clinical condition, liver function, Child-Pugh cirrhosis class, and/
or kind of surgical treatment?'*
Although there have been many studies carried out on prognostic
1196
CANCER September 15,1996 / Volume 78 / Number 6
TABLE 1
Clinical Features of 93 Cirrhotic Patients with HCC
Variable
Sex
Male
Female
Alcohol abuse
Present
Absent
HbsAg
Positive
Negative
Child-pugh class
A
B
C
Esophageal varices
Present
Absent
Pattern of the cirrhosis
Macronodular
Microimacronodular
Micronodular
Diffuse fibrosis
TABLE 2
Comparison of Different Pathologic Features of HCC
and DNA Content
No.
Variable
a2
11
88.2%
11.8%
59
34
63.4%
36.6%
23
70
24.7%
75.3%
59
30
4
63.4%
32.2%
4.4%
42
51
45.2%
54.9%
1
1.1%
79.6%
6.4%
12.9
74
6
12
HCC:hepatocellular carcinoma: HbsAg: hepatitis B surface antigen.
No. of nodules
Single
Multiple
Maximal tumor size”
5 3 cm
>3 cm
Distance from the cut level
4 0 mm
210 mm
Macroscopy
Expanding
Spreading
Diffuse
Mixed
Microscopy
Trabecular
Compact
Polymorphous
Endmondson classification
I
I1
111
N
significance of DNA flow cytometry of HCC,9,’3to the
best of our knowledge there are none on the DNA
analysis of cirrhotic parenchyma around the HCC.
Only Anti et al.I4 looked at flow cytometry in needle
biopsies in a small group of patients with hepatic lesions (some with HCC),although they did not correlate
their findings with survival.
Therefore, our first objective in this study was to
evaluate whether DNA flow cytometric analysis and
the S-phase fraction of the cirrhotic liver around HCC
nodules may predict survival in surgically treated patients.
Portal invasion
Present
Absent
Vascular invasion
Present
Absent
Necrosis
Present
Absent
u-feroprotein
SMedian
>Median
No.
Diploid
Ploidy
aneuploid
Tetraploid
34
59
11 33.3%
22 66.7%
17 35.4%
31 64.6%
6
6
50.
50.
29
64
14 42.4%
19 57.6%
8
16.7%
40 83.3%
7
5
58.
41.
47
46
16 48.5%
17 51.5%
25 52.1%
23 47.9%
6
6
50.
50.
61
19
2
7
23
4
1
1
32
10
1
2
6
5
0
1
50.
41.
91
1
1
31 96.9%
22
52
18
1
8
16
8
1
22
71
79.3%
13.7%
3.5%
3.5%
71.2%
22.2%
2.2%
4.4%
8.
48 98.0%
1 2.0%
0 -
12 100
0
0
12
30
6
0
25.0%
62.5%
12.5%
-
2
6
4
0
16.
50.
33.
8 24.2%
25 75.7%
10 79.2%
38 20.8%
4
8
33.
66.
42
51
19 57.6%
14 42.4%
27 56.2%
21 43.8%
5
7
41.
58.
38
55
16 48.5%
17 51.5%
29 60.4%
19 39.6%
10 8.
2 16.
47
46
19 57.6%
14 42.2%
21 43.7%
2 i 56.2%
7
5
0
-
1 3.1%
24.2%
48.5%
24.2%
3.1%
58.
41.
‘2‘: 10.78
P < 0.01.
MATERIALS AND METHODS
Patients
Ninety-three cirrhotic patients were studied. All patients had undergone curative hepatectomy for HCC
between August 1983 and July 1993 at the “Umberto
I st.” Hospital Turin, Italy. Curative hepatectomy was
defined as no visible tumor and/or tumor demonstrable during an intraoperative examination in the residual liver. The surgical procedures applied were anatomical resections in 77 patients (82.8%)and nonanatomical resections in 16 (17.2%).Two tumors (2.1%)
had been treated with chemoembolization before surgery. In five patients the histograms were unsuitable
for analysis and were, therefore, excluded from this
study. Histologic examination of the tumor and liver
parenchyma with cirrhosis was performed on surgical
specimens embedded in paraffin after fixation in formalin for all patients. All cases were evaluated macroand microscopically according to the main criteria described by Japanese a~thors’~-’’
and graded according
to Edmondson and Steiner.”
The following variables were analyzed: age of patient at the time of diagnosis, alcohol abuse, hepatitis
B surface antigen (HBsAg),Child-Pugh’s class, esophageal varices, cirrhosis pattern, number of nodules,
maximal tumor size, tumor distance from the margin
of resection, macroscopic pattern, Edmondson-Stein-
DNA Content and Prognosis in HCC1Rua et al.
TABLE 3
Ploidy of Cirrhotic Parenchymal Cells around HCC
Dip1oid
Aneuploid
Tetraploid
87.7%
11.2%
1.1%
81
11
1
HCC: liepaiacellular carcinoma
TABLE 4
Comparison between S-Phase Fraction and Other Prognostic
Variables in 82 Cases of Diploid Cirrhotic Liver
S-phase fraction
Variable
Modality
No. (%)
>4%
No. (%)
Age
4 2
>62
4
B t C
15 (36)
28 (70)
33 (63)
10 (331
27 (64)
12 (30)
19 (37)
20 1671
54%
Child
x2 = 9.70
P < 0.01
x2 = 6.90
P < 0.01
er’s grading, portal and/or vascular invasion, necrosis,
a-fetoprotein (AFP),nuclear DNA content of both tumor cells and of cirrhotic liver cells, and survival. HCV
testing performed after 1990 was performed on 18 patients.
Flow Cytornetry
Flow cytometry was performed on tumor and cirrhotic
liver speciniens taken from the same paraffin block,
including at least 5 to 10% of leukocytes andlor fibrocytes which were used as internal diploid reference
cells. DNA content analysis was carried out on paraffin
embedded specimens, from which 100 p thick sections
were cut immediately adjacent to those used for histology.” Sections were dewaxed, rehydrated and, after
treatment in 0.5% pepsin solution and washing,
stained with propidium iodide according to a previously described method.20-zzTo avoid cell clumping,
measurement samples were first checked at light microscopy. The measurements were performed using a
FACScan flow cytometer (Becton Dickinson, San Jose,
CAJ with a 488-nm argon ion laser.
Histograms were divided into two groups, diploid,
i.e., one GOlGl peak with a second which represented
the G2lM-phase relative to the first, and nondiploid,
i.e., those presenting different peaks, according to
Joensuu and Klemi.23
A DNA index (DI) was calculated by dividing the
modal channel number of the peak with higher DNA
content by that of the peak with lower DNA content.
1197
Peaks determining a DI value between 1.85 and 2.15
were considered to be in the G2/M region. Cases with
a peak in the G2/M region and a number of cells
greater than 15%24were considered to be tetraploid.
In diploid cases the S-phase fraction, the G2/Mphase fraction, and the proliferative index (sum of S
and G2lM fractions) were calculated from DNA histograms by DNA cell-cycle analysis software (Becton
Dickinson).
Data Analysis
The following parameters were considered for analytical purposes: sex, age, ploidy (hepatoma and cirrhosis), S-phase fraction (hepatoma and cirrhosis), maximum tumor size, number of intrahepatic nodules, portal invasion, vascular invasion, tumor distance from
the margin of resection, HBsAg, AFP, Child-Pugh’s
classification. Survival time was the dependent variable.
Survival curves were assessed according to the
Kaplan-Meier methodz5using the PROC LIFETE5T of
SAS.26The curves were plotted at monthly intervals.
Differences among curves were assessed according to
the log-rank test.27
To control the effect of potential confounders, a
multivariate analysis based on Cox’s proportional hazards regression model2*was performed on cases of
diploid cirrhosis where the S-phase fraction was evaluable. The variables analyzed included age ( 5 62 and
> 62 yrs), sex, S-phase fraction ( 5 1%,> 4%), tumor
distance from the margin of resection (> 10 mm, <
= 10 mm), number of intrahepatic nodules (single,
multiple), portal invasion, vascular invasion, ChildPugh’s classification (2 levels: A and B + C), AFP ( 5
25, > 25 ng/mL), and necrosis. The exponentiation of
the coefficient estimated from the regression model
can be expressed as relative risk (RR) of dying during
the follow-up period of subjects in the exposed category of each variable, compared with the reference
category, after allowing for the other factors entered
in the model. The PROC PHREG of SAS was used for
Cox r e g r e s ~ i o nNinety-five
.~~
percent confidence intervals (95%CI) of RRs were calculated by exponentiating
the estimated coefficient plus or minus 1.96 times its
standard error.
RESULTS
The principal clinical characteristics of the patients
were studied (Table 1). All patients were Italian, 11
women (11.8%)and 82 men (88.2%),age 26 to 78 years
(median: 62 yrs).
1198
CANCER September 15,1996 / Volume 78 / Number 6
TABLE 5
Kaplan-Meier's Estimates of the Cumulative Proportion of Survival (and Corresponding Standard Errors) according to the Considered
Prognostic Factors
Mos from surgery
Variable
Modality
No.
12
36
96
F
Sex
Male
Female
562 yrs
72
11
39
44
26
57
31
52
0.71 (0.05)
1
0.62 (0.08)
0.86 (0.05)
0.88 (0.05)
0.68 (0.06)
0.90 (0.05)
0.65 (0.07)
0.42 (0.06)
0.51 (0.16)
0.23 (0.07)
0.62 (0.08)
0.52 (0.10)
0.36 (0.07)
0.68 (0.09)
0.29 (0.06)
0.27 (0.05)
0.51 10.16)
0.16 (0.07)
0.47 (0.12)
0.42 (0.10)
0.24 (0.11)
0.51 (0.16)
0.21 10.06)
0.20
21
46
16
65
18
51
32
0.90 (0.06)
0.74 (0.06)
0.56 (0.12)
0.77 (0.05)
0.67 (0.11)
0.82 (0.05)
0.63 (0.09)
0.56 (0.11)
0.44 (0.08)
0.25 (0.11)
0.49 (0.07)
0.17 (0.09)
0.57 (0.07)
0.22 (0.08)
0.45 10.14)
0.32 (0.08)b
43
40
47
36
61
22
23
41
58
25
39
44
18
21
72
11
39
33
0.77 (0.06)
0.73 (0.07)
0.85 (0.05)
0.61 (0.08)
0.77 (0.05)
0.63 (0.10)
0.79 (0.06)
0.71 (0.07)
0.86 (0.05)
0.48 (0.10)
0.85 (0.07)
0.66 (0.06)
0.88 (0.08)
0.81 (0.09)
0.75 (0.05)
0.73 (0.13)
0.90 (0.05)
0.58 (0.09)
0.56 (0.08)
0.33 (0.08)
0.60 (0.08)
0.22 (0.07)
0.52 (0.07)
0.18 (0.09)
0.60 (0.08)
0.26 (0.08)
0.57 (0.07)
0.12 (0.07)
0.49 (0.10)
0.39 (0.07)
0.59 (0.13)
0.44 (0.12)
0.44 (0.06)
0.40 (0.16)
0.54 (0.08)
0.33 (0.08)
Age
>62 yrs
Tumor size
5 3 cm
>3 cm
Number of nodules
Edmonson-Steiner
classification
Portal invasion
Necrosis
Distance from the margin
of resection
Vascular invasion
HBsAg
a-fetoprotein
Single
Multiple
I
I1
111-IV
Absent
Present
Absent
Present
210 mm
<10 mm
Absent
Present
Negative
Positive
525 ng/mL
>25 n g h L
Child's score
A
Hepatoma's ploidy
B-C
Diploids t Tetra
Aneuploids
Hepatoma's S-phase'
55%
>5%
Cirrhosis ploidy
Diploids t Tetra
Aneuploids
Cirrhosis' S-phase'
54%
>4%
0.01
0.24
0.01
0.02
0.01
0.35 (0.10)
0.17 (0.09)
0.45 (O.ll)h
0.01
0.47 (O.llih
0.01
0.43 (0.12)
0.19 10.07)
0.40 (0.10)
0.06 (0.06)
0.42 (0.12)
0.23 (0.08)
0.41 (O.lO)b
0.01
0.39 (0.10)
0.28 (0.08)
0.53 (0.13)b
0.19
0.32 (0.08)h
0.88
0.47 (0.08)
0.13 (0.10)
0.01
0.01
0.01
0.01
0.16
Sunjival analysis performed on 83 of93 patients (10 died in the postoperative period.
"Log rank test.
" N o patient reached 96 months of follos-up.
' Only diploids.
HBsAg: heoatitis B surface antieen.
Flow Cytometry
The examined HCC were 35.5% diploid tumors, 46.2%
aneuploid, and 12.9% tetraploid; 5.4% HCC showed a
mosaic DNA distribution of aneuploidy. In the statistical analysis, 12 cases of HCC, which showed a DNA
index of 2 (tetraploid tumor) were included in the diploid group.
The DNA content of HCC was compared with conventional prognostic variables (Table 2). Tumor size
was significantly related to aneuploidy. In fact, 40 of
64 lesions larger than 3 cm were aneuploid (62.5%),
whereas only 8 of 29 tumors measuring < 3 cm were
aneuploid (27.5%) ( P < 0.01). There was no significant
relationship between ploidy, patient's age, sex, HBsAg,
AFP, Child-Pugh class, number of intrahepatic nodules, portal invasion, vascular invasion, or necrosis.
Moreover, there was not any apparent correlation between ploidy and HCV testing in the group of 18 patients evaluated.
The majority of analyzed cases of peritumoral cir-
DNA Content and Prognosis in HCC/Rua et al.
1
1199
S-phase fraction of cirrhotic cells and patient's age,
sex, HBsAg, AFP, tumor size, Kanai and Edmondson
classification, number of intrahepatic nodules, portal
invasion, vascular invasion, and necrosis.
I
-;
0
12
24
36
48
60
72
84
96
108
120
Months h r n surgery
FIGURE 1. Kaplan-Meier's estimates of the cumulative curve of survival
after hepatectomy of all the 83 patients considered in the univariate analysis.
0
12
24
36
48
60
72
84
96
108
I
120
Months h
o
r
n surgery
FIGURE 2. Cumulative proportion of survival after hepatectomy in relation
to S-phase fraction of cirrhotic parenchymal cells. A significant difference in
survival between the curves was found ( P < 0.01 by the log rank test).
rho tic tissue were diploid (87.7%),11 (11%)were aneuploid, and just 1 was tetraploid (Table 3). The diploid
cases were selected to compare their S-phase fraction
with two other important prognostic factors such as
age and Child-Pugh classification. When S-phase fraction was compared with age, 64% of the patients
younger than 62 years (64%) showed high S-phase
value (>4%), whereas only 30% of the older patients
showed the same values ( P < 0.01). Child-Pugh classes
were significantly related to S-phase fraction of cirrhotic liver, in fact, 36.5% of patients with Child-Pugh
Class A and 66.7% of patients with Child-Pugh Class
B or C showed a high S-phase fraction ( P < 0.01) (Table 4). There was no significant relationship between
Survival analysis
Analysis was made on 83 cases because 10 patients
(10.7%)who died in the postoperative period (within
2 mos) were excluded from the analysis of survival.
Follow-up was continued until December 1994 or until
a patient's death and ranged from 2 to 120 months
(median, 30). Forty-nine patients died of neoplastic
disease.
The univariate analysis of survival showed that
sex, tumor size, hepatoma S-phase fraction, hepatoma, and cirrhosis ploidy were not significantly correlated with survival (Table 5). The cumulative proportion of survival was 0.31 with 5 patients still alive 96
months after diagnosis (Fig. 1).Cumulative proportion
of survival in relation to S-phase fraction of diploid
cirrhotic liver was significantly higher in low (54%)
versus high (> 4%) S-phase fraction cases ( P < 0.01
by the log-rank test) (Fig. 2).
Cox Proportional Hazard Model
Multivariate analysis demonstrated that the most predictive variables in decreasing order were Child-Pugh
score, S-phase fraction of cirrhotic liver, and presence
of necrosis. Conversely, significant prognostic indicators by univariate analysis, such as age, number of
intrahepatic nodules, portal invasion, vascular invasion, HbsAg and AFP were unsignificant by multivariate analysis (Table 6).
DISCUSSION
In this study we demonstrated that the S-phase fraction of cirrhotic liver with diploid DNA content might
predict the survival of patients with HCC. In our series
the cirrhotic liver around HCC nodules was diploid in
91.1% of the cases, aneuploid in 7.8%, and tetraploid
in 1.1%.The DNA analysis of the cirrhotic parenchyma
showed that polyploidization was dramatically reduced in patients with HCC and aneuploid clones
were present among diploid cells. It is known that
normal human liver contains polyploid hepatocytes,
and that the majority of them have two diploid nuclei.
Polyploidization is thought to reflect increasing degrees of irreversible hepatocellular differentiati~n.~'
In
contrast, significantly elevated fractions of diploid hepatocytes and a reduction in the polyploid populations
have been reported in humans and experimentally induced hepatocellular carcinoma^.^' Moreover in liver
affected by chronic hepatitis and cirrhosis the diploid/
1200
CANCER September 15,1996 / Volume 78 / Number 6
TABLE 6
Association between Prognostic Factors and Probability of Death
Variable
Modality
RR
Sex
Male
Female
562 yrs
>62 yrs
Negative
Positive
A
B-C
Single
Multiple
1.00
1.50
1.00
1.22
1.00
2.39
1.oo
3.08
1.00
0.77
210 mm
<10 mm
1.00
1.82
1.00
2.94
1.00
1.93
1.00
1.59
1.00
2.00
1.oo
3.61
Age
HBsAg
Child
Number of nodules
Distance from the margin
of resection
Necrosis
cu-fetoprotein
Vascular invasion
Portal invasion
S-phase of cirrhosis
Absent
Present
525 n g h L
>25 ng/mL
Absent
Present
Absent
Present
54%
>4%
CI
Chi-square
-
0.47-4.8
0.47
-
0.46-3.2
0.16
-
0.98-5.80
3.69
-
1.44-6.62
8.32
0.23-2.52
0.19
-
0.79-4.15
1.98
-
1.32-6.57
6.94
-
0.95-3.93
3.28
-
0.61-3.93
1.60
-
0.68-5.82
0.90
-
1.51-8.63
8.28
RR: relative risk; CI: 95% confidence interval; HBsAg: hepatitis B surface antigen.
The degree of association is expressed as RR and corresponding CI estimated fitting Cox multivariate proportional hazard model that includes as covariates gender
and factors significantly associated with survival by univariate analysis (Table 51,
polyploid ratios increased, whereas the hepatocyte binuclearity decreased this trend was even more
marked in euploid nodules (premalignant and malignant), in which the S-phase fraction was significantly
greater than that of the normal 1 i ~ e r . l ~
On the grounds of the hypothesis of a relationship between the development of carcinoma and an
increased cell proliferation, presumably by an increased rate of random mutations,’ we analyzed the
follow-up of patients with HCC associated to diploid
cirrhotic liver in relation to high and low S-phase.
In effect, the survival of patients with HCC which
developed in a cirrhotic liver with a diploid DNA
content and a high S-phase fraction was significantly
reduced.
In agreement to our findings, Tarao et al.’ demonstrated that HCC developed in a 3-year period in
64.3% of the cirrhotic patients with high-DNA synthesis activity, and only in 14.3% of the cirrhotic
patients with low-DNA synthesis activity. Ballardini
et
found higher reactivity for proliferating cell
nuclear antigen in the cirrhotic liver of patients who
eventually developed HCC (median: 0.7 vs. 2.61,
therefore they suggest a differentiated follow-up of
these patients.
To our knowledge no data are reported about
survival of patients with HCC and proliferation of
hepatic cells in the peritumoral cirrhotic liver. Jwo
et al.” showed that patients with HCC > 5 cm, or
with multipleldaughter HCC nodules had a shortened disease free survival. In the same work patients
with diploid or aneuploid pattern but with single
GOlGl peak had a reduced overall survival. Other
authors showed that patients with aneuploid HCC
had a significantly worse prognosis than did those
with diploid HCC, also in groups subdivided according to tumor size. Among DNA aneuploid patients, the survival times were significantly less for
patients with a low DNA index (<1.5) than for those
with a high DNA index ( ~ 1 . 5 ) . ’
By univariate analysis we could not demonstrate
a significant correlation of HCC ploidy with survival,
whereas we found significant roles for age, number
of intrahepatic nodules, Edmondson and Seiner’s
classification, portal invasion, vascular invasion, tumor distance from margin of resection, presence of
DNA Content and Prognosis in HCC/Rua et al.
necrosis, HbsAg, AFP, Child-Pugh’s score, and Sphase fraction of cirrhotic cells. In analogy to our
results, a significant association between survival
rates and the presence of vascular invasion or intrahepatic metastasis was found in the study of Hamazaki et al.32 and significant association was found
between DNA ploidy and HCC prognosis.
However, because univariate analysis does not always reflect the actual significance of a prognostic factor, multivariate analysis and calculated cumulative
proportion of survival, in relation to clinicopathologic
factors, were performed. The Cox’s multivariate survival analysis showed that only Child-Pugh’s score, Sphase fraction of diploid cirrhotic cells, and necrosis
were statistically significant independent prognostic
factors. Using the same multivariate analysis with
Cox’s proportional hazards model, Okada” showed
that DNA diploidy, absence of both alcohol abuse and
vascular invasion, were independent favorable predictive factors, whereas Fujimoto et al.,’ showed that
nuclear DNA content provided significant prognostic
value, as did vascular invasion and intrahepatic metastasis.
In our study, together with high S-phase fraction
of cirrhotic liver parenchyma, Child-Pugh classification was another independent parameter influencing
tumor behavior. This can be justified by the impaired
function of the liver, after resection of the tumor.
In conclusion, our data suggest that S-phase fraction of cirrhotic liver parenchyma may be employed
as a new additional parameter in the prognostic evaluation of HCC patients.
REFERENCES
1. Tarao K, Ohkawa S, Shimizu A, Harada M, Nakamura Y, Ito
Y, et al. Significance of hepatocellular proliferation in the
development of hepatocellular carcinoma from anti-hepatitis C virus-positive cirrhotic patients. Cancer 1994;73:114954.
2. Ballardini G, Groff P, Zoli M, Hianchi G, Groff P, Zoli M,
Bianchi G, Giostra F, Francesconi €1, et al. Increased risk
of hepatocellular carcinoma development in patients with
cirrhosis and with high hepatocellular proliferation. J Hepato1 1994;20:218-22.
3. Arakawa M, Kage M, Sugihara S, Nakashima T, Suenaga M,
Okuda K. Emergence of malignant lesions within an adenomatous nodule in cirrhotic liver: observation in five cases.
Gastroenterology 1988;95:1664-6.
4. Sakamoto M, Hirohashi S, Shimosato Y. Early stages of
multistep hepatocarcinogenesis adenomatous hyperplasia
and early hepatocellular carcinoma. Hum Puthol 1991;
22: 172-8.
5. Lai CI,, Wu PC, Lam KC, Todd D. Histologic prognostic indicators in hepatocellular carcinoma. Cancer 1979;44:167785.
6. Franco D, Capussotti L, Smadja C, Bouzari H, Meakins J ,
Kemeny F , et al. Resection of hepatocellular carcinomas.
1201
Results in 72 european patients with cirrhosis. Gastroenterology 1990;98:733-8.
7. Yamanaka N, Okamoto E, Toyosaka A, Mitunobu M, Fujihara S, Kato T, et al. Prognostic factors after hepatectomy
for hepatocellular carcinomas. A univariate and multivariate
analysis. Cancer 1990;65:1104- 10.
8. Shirabe K, Kanematsu T, Matsumata T, Adachi E, Akazawa
K, Sugimachi K. Factors linked to early recurrence of small
hepatocellular carcinoma after epatectomy: univariate and
multivariate analyses. Hepatology 1991;14:802-5.
9. Fujimoto J, Okamoto E, Yamanaka N, Toyosaka A, Mitsunobu M. Flow cytometric DNA analysis of hepatocellular
carcinoma. Cancer 1991;67:939-44.
10. Arii S, Tanaka J, Yamazoe Y, Minematsu S, Morino T, Fujita
K, et al. Predictive factors for intrahepatic recurrence of hepatocellular carcinoma after partial hepatectomy. Cancer
1992;69:913-9.
11. Jwo SC, Chiu JH, Chau GY, Loong CC, Lui WY. Risk factors
linked to tumor recurrence of human hepatocellular carcinoma after hepatic resection. Hepatology 1992; 16:1367(1.
12. Okada S, Shimada K, Yamamoto J, Takayama T, Kosuge T,
Yamasaki S, et al. Predictive factors for postoperative recurrence of hepatocellular carcinoma. Gastroenterology 1994;
106:1618-24.
13. Cong WM, Wu MC. The biopathologic characteristics of
DNA content of hepatocellular carcinomas. Cancer 1990;
66~498-501.
14. Anti M, Marra G, Rapaccini GL, Rumi C, Bussa S, Fadda
G, et al. DNA ploidy pattern in human chronic liver diseases and hepatic nodular lesions. Flow cytometric analysis on echo-guided needle liver biopsy. Cancer 1994;73:
281-8.
15. Okuda K, Ohtsuki T, Obata H, Tomimatsu M, Okazaki N,
Hasegawa H, et al. Natural history of hepatocellular carcinoma and prognosis in relation to treatment. Study of 850
patients. Cancer 1985;56:918-28.
16. Kanai T, Hirohashi S, Upton MP, Noguchi M, Kishi K, Makuuchi M, et al. Pathology of small hepatocellular carcinoma.
A proposal for a new gross classification. Cancer 1987;
6018 10- 9.
17. Nakashima S, Okuda K, Kojiro M, Jimi A, Yamaguchi R, Sakamot0 K, et al. Pathology of hepatocellular carcinoma in Japan. 232 consecutive cases autopsied in ten years. Cancer
1983;51:863-77.
18. Edmonson HA, Steiner PE. Primary carcinoma of the liver.
A study of 100 cases among 48,900 necropsies. Cancer
1954;71462-503.
19. Stephenson RA, Gay H, Fair WR. Melamed MR. Effect of
section thickness on quality of flow cytometric DNA content
determinations in paraffin-embedded tissue. Cytometry
1986;7:4 1-4.
20. Coon JS, Landay AL, Weinstein RS. Flow cytometric analysis
of paraffin-embedded tumors: implications for diagnostic
pathology. Hurn Pnthol 1986;117:435-7.
21. Hedley DW, Friedlander ML, Taylor JW, Taylor IW, Rugg CA,
Musgrove EA. Method for analysis of cellular DNA content
of paraffin-embedded pathological material using flow cytometry. J Histocliem Cytochein 1983;31:1333-5.
22. Hedley DW, Friedlander ML, Taylor IW. Application of DNA
flow cytometry to paraffin-embedded archival material for
the study of aneuploidy and its clinical significace. Cytonietry 1985;6:327-33.
1202
CANCER September 15,1996 I Volume 78 I Number 6
23. Joensuu H, Klemi PJ. Comparison of nuclear DNA content
in primary and metastatic differentiated thyroid carcinoma.
Am J Clin Puthol 1988;89:35-40.
24. Kallioniemi OP, Blanco G, Alavaikko M, Hietanen T, Mattila
J, Lauslahti K, et al. Improving the prognostic value of DNA
flow cytometry in breast cancer by combining DNA index
and S-phase fraction: a proposed classification of DNA histograms in breast cancer. Cancer 1988;62:2183-90.
25. Kaplan EL, Meier P. Nonparametric estimation from incomplete observations. JASA 1958;53:457-81.
26. SAS Institute Inc. SAS/STAT User’s Guide. Version 6 4 t h ed.,
Cary (NC): SAS Institute Inc., 1989.
27. Mantel N. Evaluation of survival data and two new rank
order statistics arising in its consideration. Cancer Chemother Rep 1966;50:163-70.
28. Cox DR. Regression models and life tables. J R Stat SOC
1972;B33: 187-220.
29. SAS Technical Report P-229 SAS/STAT software. Changes
and enhancements. Release 6.07. 1992:433-79.
30. Saeter G, Lee CZ, Schwarze E, Ous S, Chen DS, Sung JL, et
al. Changes in ploidy distributions in human liver carcinogenesis. JNCI 1988;80:1480-5.
31. Scott RJ, Chakraborty S, Sell S, Hunt JM, Dunsford HA.
Change in the ploidy state of rat liver cells during chemical
hepatocarcinogenesis and its relationship to the increased
expression of a-fetoprotein. Cuncer Res 1989;49:608590.
32. Hamazaki K, Kato T, Yunoki Y, Mori M, Gochi A, Mimura
H, et al. An analysis of DNA ploidy pattern of hepatocellular
carcinoma. Acta Med Okayumu 1993;47:413-6.
Документ
Категория
Без категории
Просмотров
4
Размер файла
602 Кб
Теги
620
1/--страниц
Пожаловаться на содержимое документа