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Int. J. Cancer (Pred. Oncol.): 79, 116–120 (1998)
r 1998 Wiley-Liss, Inc.
Publication of the International Union Against Cancer
Publication de l’Union Internationale Contre le Cancer
DNA PLOIDY AND CELL-CYCLE ANALYSIS IN INTRACRANIAL MENINGIOMAS
AND HEMANGIOPERICYTOMAS: A STUDY WITH HIGH-RESOLUTION
DNA FLOW CYTOMETRY
Anton ZELLNER1, Jürgen MEIXENSBERGER2*, Wolfgang ROGGENDORF3, Michael JANKA2, Holger HOEHN4 and Klaus ROOSEN2
of Neurology, University of Würzburg, Würzburg, Germany
2Department of Neurosurgery, University of Würzburg, Würzburg, Germany
3Department of Pathology, University of Würzburg, Würzburg, Germany
4Department of Genetics, University of Würzburg, Würzburg, Germany
1Department
Although various DNA flow-cytometric studies have been
performed on meningiomas, the role of DNA ploidy and the
S-phase fraction (SPF) in predicting biological tumor behavior remains unresolved. Discrepant results in earlier studies
might be due to different preparing, staining and measuring
techniques; different quality standards; and lack of sophisticated computer software. In this study, high-resolution DNA
flow cytometry using the DNA-specific dye DAPI (48, 68diamidino-2-phenylindol) was performed on stored frozen
tissue from 128 microsurgically resected meningiomas and 7
hemangiopericytomas, including 17 recurrent meningiomas
and 4 recurrent hemangiopericytomas. The computer software Multicycle 2.5 was used to determine the ploidy level
and to perform cell-cycle analysis. DNA aneuploidy and SPF
were significantly higher in atypical, anaplastic and recurrent
meningiomas and correlated well with histopathological features such as focal necrosis, infiltration of dura mater and
mitotic activity. Among 128 meningiomas, 42 had additional
DNA aneuploid stem lines. No association between hypo- and
hyperploidy and either histological subtype or clinical outcome was found. In 7 hemangiopericytomas, SPF was significantly higher compared to the benign meningioma group,
while only 1 tumor was aneuploid. In all 42 DNA aneuploid
tumors, cell-cycle analysis was performed separately for the
euploid and aneuploid stem lines. The proliferation parameters (SPF, G2/M phase) were significantly higher in the DNA
aneuploid stem lines. DNA ploidy and SPF are thus useful
indicators of different biological behavior within identical
histological subgroups in meningiomas. Int. J. Cancer (Pred.
Oncol.) 79:116–120, 1998.
r 1998 Wiley-Liss, Inc.
Despite sophisticated histological grading systems based on
morphological and immunohistochemical parameters, it is not
always possible to predict the individual proliferative potential and
tendency of recurrence among tumors within the same histological
subtype; this is particularly true for low- and middle-grade tumors
(Coons et al., 1994; Ironside et al., 1987; May et al., 1989).
Thus, even among meningiomas that are histologically classified
as benign and completely resected by microsurgery, the number of
tumor relapses varies between 5% and 20%.
Flow-cytometric parameters, such as DNA ploidy and S-phase
fraction (SPF), were significantly associated with tumor behavior
in many tumor entities, such as breast cancer, non-small cell lung
cancer, ovarian carcinoma and colorectal carcinoma (Coon et al.,
1987; Giaretti and Santi, 1990; Merkel and McGuire, 1990;
Seckinger et al., 1989). Although many studies have been performed on meningiomas and neuro-epithelial tumors, the role of
DNA ploidy and SPF in these tumor groups remains unresolved
(Rabson, 1994).
Discrepant results might be due to technical reasons (e.g.,
sample preparation, staining) or variable definitions (e.g., definition
of DNA aneuploidy and DNA index). In addition, many studies do
not incorporate internal cell standards or achieve only poor
coefficients of variation (CV) in their measurements (Dressler,
1990; Hiddemann et al., 1984; McCarthy and Fetterhoff, 1989;
Shapiro, 1989). Finally, in earlier studies, sophisticated computer
software was not available to perform cell-cycle analysis simultaneously in different stem lines of the same tumor sample.
The present study aimed at avoiding these problems by using
high-resolution DNA histograms the sophisticated Multicycle 2.5
software. Our prospective study seeks to answer the following
questions:
1. Do flow-cytometric DNA parameters such as ploidy and SPF
have prognostic relevance in predicting an aggressive tumor
behavior?
2. Can these parameters identify different prognostic groups
within histopathologically graded and defined tumor subgroups?
3. Is there a significant difference in the proliferative activity
between DNA diploid and DNA aneuploid tumor stem lines?
MATERIAL AND METHODS
We studied 135 samples from complete microsurgically resected
tumors that were frozen in liquid nitrogen immediately after
surgery. Our protocol was approved by the local ethics committee.
Tissue samples were stored at 280°C for an average of 6 months
prior to flow-cytometric analysis. Table I summarizes essential data
and histological grading (WHO) of the tumor material.
Immediately after thawing, tumor samples were minced and
prepared as single-cell-nuclei suspensions using citric acid and
Tween 20 (Sigma, Deisenhofen, Germany) as detergent according
to the modified protocol of Otto (1990). Cell nuclei were stained
with the AT-specific dye DAPI (48, 68-diamidino-2-phenylindol)
(Sigma). Chicken erythrocytes and calf thymocytes were treated
like tumor tissue and were used as internal DNA standards. In about
40 tumors, we prepared and measured an average of 3 samples
within different areas of the same tumor in order to evaluate tissue
heterogeneity.
All flow-cytometric measurements were performed on a PAS II
flow cytometer (Partec, Reinach, Switzerland) using a mercury arc
lamp as light source. More than 400 measurements with an average
of 19,500 signals (cell nuclei) counted in each sample were
evaluated. Mean values for nuclear aggregation (clumping) were
6.14 6 3.54% and for debris 7.54 6 8.19%. The resulting DNA
histograms (Fig. 1) were evaluated with the computer software
Multicycle 2.5 (Phoenix Flow Systems, Seattle, WA; Rabinovitch).
Histological classification was performed according to the WHO
classification for meningiomas (Kleihues et al., 1991). Simultaneously, a qualitative and semi-quantitative morphological investigation concentrated on the number of mitoses, infiltrative growth,
tightness and pleomorphism of cells, as well as tumor necrosis.
*Correspondence to: Department of Neurosurgery, Universitätsklinik
Würzburg, Josef-Schneiderstrasse 11, D-97080 Würzburg, Germany. Fax:
011-49-931-2012635. E-mail: [email protected]
Received 8 August 1997; Revised 17 October 1997
DNA FLOW CYTOMETRY IN MENINGIOMAS AND HEMANGIOPERICYTOMAS
TABLE I – CLINICAL DATA AND HISTOLOGICAL GRADING (WHO)
ON THE TUMOR MATERIAL
Histology
Meningioma subtype
Meningothelial
Transitional
Fibroblastic
Psammomatous
Microcystic
Secretory
Myxomatous
Atypical
Anaplastic
Hemangioblastic
Hemangiopericytomas
Total case number
Cases
(n)
WHO
grade
Age
(mean,
years)
Sex
ratio
(m:f)
Recurrent
tumors
(n)
39
36
22
2
3
2
1
17
5
1
7
135
1
1
1
1
1
1
1
2
3
2
2
57
61
61
60
44
58
42
61
63
52
52
11:28
10:26
1:21
1:1
1:2
0:2
1:0
3:14
3:2
0:1
2:5
5
3
0
0
0
0
0
7
2
0
4
21
TABLE II – DISTRIBUTION OF ANEUPLOID TUMORS AND PERCENTAGES
OF THE ANEUPLOID POPULATION (% ANTOT) IN ANEUPLOID TUMORS
Histology
Cases Aneuploid
(n) tumors (n)
Meningioma subtype
Meningothelial
39
Transitional
36
Fibroblastic
22
Psammomatous
2
Microcystic
3
Secretory
2
Myxomatous
1
Atypical
17
Anaplastic
5
Hemangioblastic
1
Hemangiopericytomas
7
Total case number
135
11
9
0
0
3
0
1
12
5
1
1
43
% Antot
(mean 6 SD)
Recurrent
tumors (n)
53.55 (622.17)
65.04 (611.75)
—
—
70.45 (62.95)
—
51.2
67.93 (617.63)
77.42 (610.81)
54.3
Not evaluable
63.97 (618.43)
5
3
0
0
0
0
0
7
2
0
4
21
117
S phase
SPF indicates how many cells of the entire population are
replicating DNA at a given time point. SPF is computed by the
polynomial S-phase fitting method described by Dean and Jett
(1980).
DNA ploidy level
The relation of the tumor stem cell peak to the reference peaks
(chicken and calf thymocytes) is defined as the DNA index (DI)
and relates to the DNA-specific fluorescence emitted by a given
cell population. By definition, the DI for the diploid G0/G1 peak is
set at 1, the DI for the G2/M peak is 2 and additional aneuploid stem
lines have a DI ,1 (hypodiploid) or .1 (hyperdiploid). In our
series, the existence of additional DNA aneuploid stem lines was
accepted only when 2 clearly visible and distinct (G0/G1 ) peaks,
which could be reproduced in at least 2 independent measurements,
were demonstrated in the histograms.
Identification and calculation of tetraploid stem lines is more
difficult because the tetraploid G0/G1 peak occupies the same
position in the histogram as the G2/M peak of the diploid stem line.
In our study, tetraploidy was considered when the G2/M phase of
the diploid stem line exceeded 10% of all measured cell nuclei and
a corresponding G2/M phase of the presumptive tetraploid stem
line was evident.
Coeffıcient of variation
The CV is used to describe the width of the peaks in the DNA
histograms. It is a normalized standard deviation defined as CV 5
100 times SD divided by the mean of the peak.
Statistical analysis
Results are given as mean 6 SD for the different groups.
Statistical analysis was performed using the x2 test, t-test, Wilcoxon test and variance analysis, as appropriate.
RESULTS
FIGURE 1 – DNA histogram of a DNA euploid meningioma with
internal standard (chicken erythrocytes) evaluated by the computer
software Multicycle 2.5.
Tumor necrosis was assessed only in pre-operatively nonembolized tumor tissue.
DNA classification
The program Multicycle calculates 3 cell-cycle compartments
(G0/G1, S, G2/GM) separately for different stem lines within a
tumor sample.
As shown in Table I, 128 cases of intracranial meningiomas and
7 hemangiopericytomas were included in our study. Among these,
21 recurrent tumors could be evaluated (17 meningiomas and 4
hemangiopericytomas). The average CV for all diploid G0/G1
peaks was 2.01 6 0.58% and for all aneuploid peaks 1.86 6 0.46%.
A tumor sample was considered unusable when the CV in repeated
measurements exceeded 3.5%. This was the case in about 20 tumor
samples, which were not considered in our series; 43 of the 135
tumors had an additional DNA aneuploid stem line, which was
hypodiploid in 18 cases (41.9%) and hyperdiploid in 19 cases
(44.2%). Four tumors (9.3%) had a tetraploid stem line. In 2
tumors, either the cell-cycle parameters (case in Fig. 2b) or the DI
of the aneuploid stem line (the one aneuploid hemangiopericytoma) could not be accurately determined by the program Multicycle (one case in Fig. 2b) and, therefore, were not considered in
statistical analyses.
No association between hypo- and hyperdiploidy and either
histological subtype or tumor recurrence was noted. Concerning
the peak position of the aneuploid stem lines, there appeared to be a
tendency of the aneuploid G0/G1 peaks to be more distant from the
diploid stem line (DI 5 1) along with an increasing malignant
potential or WHO grade (Figs. 3, 4). Table II lists the distribution of
aneuploid tumors among the histological subgroups and the
relative percentages of the aneuploid tumor fraction (% Antot).
All of the 5 anaplastic meningiomas (WHO 3) and 12 of the 17
(70.6%) atypical meningiomas (WHO 2) exhibited DNA aneuploid
stem lines, while of the benign (WHO 1) meningiomas only 11 of
39 (28.9%) meningothelial, 9 of 36 (25%) transitional and 0 of 22
fibroblastic tumors were DNA aneuploid. This difference (WHO 1
vs. WHO 2/3) was highly significant ( p , 0.001).
In contrast with these results, only 1 of 7 (14.2%) hemangiopericytomas (WHO 2) showed a DNA aneuploid cell line. One
118
ZELLNER ET AL.
A
FIGURE 4 – Distribution of DNA aneuploid stem lines (according to
the DNA indices of the corresponding G0/G1 peaks) in 16 atypic and
anaplastic (WHO 2/3) meningiomas. A DNA index of 1 refers to the
position of the diploid G0/G1 peak.
B
FIGURE 5 – Among DNA aneuploid meningiomas (n 5 43, gray
and black columns), the SPF, G2/M phase and PI of the aneuploid stem
line were significantly higher compared to the diploid stem line,
***p , 0.0001. As a comparison, the mean values of DNA euploid
meningiomas (n 5 92) are indicated by the white columns.
FIGURE 2 – DNA histograms of 2 different tumor probes of the
same atypical recurrent meningioma depicting local tumor heterogeneity. While in probe 1 (a) only 1 additional hypoploid stem line could be
detected, probe 2 (b) showed 2 additional hypoploid stem lines. The
exact positions of the peaks were determined in histograms with
internal DNA standards (not shown). Cell-cycle parameters in this case
(b) were not calculated accurately by Multicycle.
FIGURE 3 – Distribution of DNA aneuploid stem lines (according to
the DNA indexes of the corresponding G0/G1 peaks) in 24 benign
(WHO 1) meningiomas. A DNA index of 1 refers to the position of the
diploid G0/G1 peak.
recurrent, atypical meningioma had 2 DNA hypoploid stem lines
(Fig. 2a,b).
Tumors with aneuploid stem lines showed a significantly higher
rate of histomorphological evidence for focal necrosis ( p , 0.01),
infiltration of dura mater ( p , 0.05) and a higher mitotic activity
( p , 0.001). No significant correlation between the percentages of
the aneuploid tumor fractions (% Antot) and parameters of
proliferation (SPF, proliferation index [PI]) was observed, although
there was an unequivocal tendency toward increased % Antot
fraction in tumors with higher rates of focal necrosis, infiltration of
dura mater and high mitotic activity.
Among the 21 recurrent tumors, 11 had additional aneuploid
stem lines (52.3%), while only 32 of the 114 primary tumors
(28.1%) were DNA aneuploid. This difference was significant
( p , 0.05).
Cell-cycle analysis
The SPF of all DNA diploid tumors (n 5 92) was compared
between the histological subtypes. The atypical meningiomas
(n 5 5, WHO 2) and the hemangiopericytomas (n 5 6, WHO 2)
showed a significantly higher ( p , 0.001) mean SPF of 2.1 6
1.4% and 1.6 6 1.6% compared to the meningothelial (0.7 6 0.6%,
n 5 28), transitional (0.6 6 0.6%, n 5 27) and fibroblastic
(0.7 6 0.6%, n 5 22) subgroups.
Among the 43 tumors with aneuploid stem lines the SPF,
G2M-phase and the PI (SPF 1 G2/M-phase) were calculated separately for the diploid stem line and the DNA aneuploid stem line
fractions. All parameters were significantly ( p , 0.001) higher in
the aneuploid stem lines (Fig. 5). Additionally, in contrast with
diploid tumors, no significant difference in the SPF between
histological subgroups was evident.
In accordance with the ploidy evaluation results, significantly
higher levels of SPF were noted in meningiomas with higher
mitotic rates ( p , 0.05), whereas tumors infiltrating the cortex or
exhibiting focal necrosis had no significantly different SPF levels.
The 21 recurrent tumors had significantly higher SPF levels than
the 114 primary tumors ( p , 0.05).
DNA FLOW CYTOMETRY IN MENINGIOMAS AND HEMANGIOPERICYTOMAS
DISCUSSION
Quality criteria
Although various previous DNA flow-cytometric studies have
been performed on meningiomas (Ironside et al., 1987; May
et al., 1989; Ahyai and Spaar, 1987; Appley et al., 1990; Assietti et
al., 1990; Butti et al., 1989; Crone et al., 1988; Cruz-Sanchez et al.,
1993; Kawamoto et al., 1989; Nishizaki et al., 1993; Spaar et al.,
1987), rather conflicting results have been reported concerning the
number of DNA aneuploid stem lines and the distribution of the
respective cell-cycle phases. The reasons for such discrepancies
might be different methods of cell preparation and differences in
instrumentation and flow cytometry, including histogram evaluation. Another explanation might be the smaller sample numbers or
differences in definition of DNA aneuploidy. Quality criteria for
flow-cytometric measurements set in earlier studies (Dressler,
1990; Hiddemann et al., 1984; McCarthy and Fetterhoff, 1989;
Shapiro, 1989) often have been ignored. In addition to parameters
such as the number of counted cells, amount of debris and cell
aggregation (clumping), CV is one of the most important criteria in
evaluating flow-cytometric DNA histograms (Dressler, 1990; McCarthy and Fetterhoff, 1989). Lower CVs imply higher precision
and a greater chance to identify DNA aneuploid stem lines within
tumor tissue. The CV of such measurements, therefore, should not
exceed 3% (Seckinger et al., 1989; Shapiro, 1989). In our study, the
average CV for all diploid G0/G1 peaks was 2.01 6 0.58% and for
all aneuploid peaks 1.86 6 0.46%. Moreover, compared to earlier
investigations, our series of 128 meningiomas comprises a very
large and homogeneous group.
The role of DNA aneuploidy as an indicator of an aggressive
biological tumor behavior is unresolved. Butti et al. (1989) and
Spaar et al. (1987) described a significantly higher rate of
aneuploid stem lines in malignant meningiomas. Crone et al.
(1988) and Ironside et al. (1987) have shown an increase in SPF
and PI in DNA aneuploid meningiomas. Cruz-Sanchez et al. (1993)
observed that DNA aneuploidy was significantly associated with
tumor recurrence. However, Mathiesen et al. (1989) and Akachi et
al. (1991) found no increased DNA aneuploidy in malignant
meningiomas and Nishizaki et al. (1990) showed that evaluating
tumor prognosis based on the presence of aneuploid stem lines
yields very uncertain estimates.
Concerning proliferation parameters like SPF and PI, many
authors agree on their significance in indicating an aggressive
biological behavior (Ironside et al., 1987; May et al., 1989; Akachi
et al., 1991; Butti et al., 1989; Crone et al., 1988; Cruz-Sanchez et
al., 1993; Finn et al., 1994) in malignant meningiomas. In our
series, only 24 of 105 (22.9%) benign (WHO 1) meningiomas were
DNA aneuploid, while 12 of 17 (64%) atypical tumors and all
malignant meningiomas (n 5 5) had additional aneuploid stem
lines. SPF was significantly higher ( p , 0.001) in atypical meningiomas compared to benign tumors. In atypical meningiomas
(WHO 2), an intermediate position between benign and malignant
tumors with respect to DNA ploidy was demonstrated here.
Both DNA aneuploidy and SPF were significantly increased in
recurrent tumors (n 5 21) compared to the primary tumor group
(n 5 114). In the fibroblastic meningioma group (n 5 22), no
aneuploid stem line and no tumor recurrence was found.
During a post-operative follow-up period ranging from 8 to 54
months (median 25 months), we observed 8 recurring tumors
among 6 different patients in our series. In every case, the primary
tumor had an aneuploid stem line, whereas none of the 92 primary
DNA euploid tumors showed evidence of recurrence.
Evaluation of tissue heterogeneity of 35 tumors with an average
of 3 different samples/tumor tested revealed no variation of DI for
both diploid and aneuploid stem lines. Except in 1 case (Fig. 2a,b),
no different aneuploid stem lines were detected in different areas of
the same tumor. However, the percentages of the aneuploid tumor
fraction (% Antot) varied up to 10% in the benign and up to 22% in
the atypical and anaplastic meningiomas. The variability of the
119
G0/G1 phase and SPF was below 5% in benign and below 10% in
atypical and anaplastic meningiomas.
In earlier flow-cytometric investigations, comparison of cellcycle data in different DNA stem lines within the same tumor was
performed rarely due to a lack of qualified computer software
(Assietti et al., 1990). In our study, among 43 DNA aneuploid
tumors, the diploid and aneuploid cell cycles were evaluated and
compared separately with the computer software Multicycle. The
SPF, G2/M phase and PI were strongly increased in the DNA
aneuploid stem lines compared with the diploid stem lines (Fig. 5).
Only 2 previous studies attempted to define hypo- and hyperploid cell lines. While Ironside et al. (1987) found 6 hypoploid and
10 hyperploid cell lines among 16 DNA aneuploid tumors,
Cruz-Sanchez et al. (1993) detected 10 hypoploid and 7 hyperploid
lines among 17 aneuploid tumors and found a slightly increased
tendency of recurrence in the hyperploid tumor group.
There is only sparse published evidence concerning the possible
relevance of either hypo- or hyperploidy for different biological
and therapeutic behaviors of intracranial tumors. Sandberg and
Turc-Carel (1987) associated chromosomal hypodiploidy with an
aggressive behavior in meningiomas. For cultivated glioma cell
lines, Shapiro and Shapiro (1984) demonstrated that hypo- or
near-diploid cell lines become more readily resistant to chemotherapy, while hyperploid cell lines are more sensitive. Similar results
are known for gliomas (Kobayashi and Hoshino, 1985), neuroblastomas (Look et al., 1984), lymphoblastic leukemia (Williams et al.,
1982) and bladder carcinomas (Quirke, 1986).
In contrast to our results, most cytogenetic studies analyzing
chromosomal patterns of intracranial tumors only occasionally
provide evidence for additional hyperploid stem lines (Heim and
Mitelman, 1987; Katsuyama et al., 1986; Maltby et al., 1988;
Westphal et al., 1989, Zang, 1981). Likewise, the hypothesis of
Poulsgard et al. (1989), that the general tendency of clonal
evolution in meningiomas goes toward hypodiploidy and that this
appears to be correlated with a more aggressive behavior of the
tumor (Sandberg and Turc-Carel, 1987), was not confirmed in our
study.
A possible explanation for these discrepant results might be
changes of the ploidy status of the tumor in cell culture (Kawamoto
et al., 1989; Coons and Johnson, 1993). Shapiro and Shapiro
(1984) described an increased instability for hyperploid glioma cell
lines compared to near- or hypodiploid cell lines. It is also true for
meningiomas that flow-cytometric data from cell culture tissue and
fresh tissue are not strictly comparable. This issue is currently
under investigation in our laboratory.
The biological behavior of hemangiopericytomas is difficult to
predict (Enzinger and Weiss, 1988). Although histomorphological
features have been used to differentiate between benign and
malignant hemangiopericytomas, it is generally assumed that many
of these tumors have an uncertain malignant potential (Finn et al.,
1994; Enzinger and Weiss, 1988). Finn et al. (1994) found no
evidence for a DNA aneuploid stem line among 22 hemangiopericytic tumors, while high SPF and PI indicated an aggressive
behavior in cases of metastasis, local recurrence following complete surgical excision or direct invasive disease. Likewise, in our
series of 7 hemangiopericytomas, only 1 tumor had a DNA
aneuploid stem line, while the SPF was significantly increased
( p , 0.001) compared to benign meningiomas. In agreement with
Finn et al. (1994), DNA ploidy does not appear to be a useful
indicator of biological behavior in hemangiopericytomas, while
cell-cycle-proliferative parameters such as SPF and PI should be
studied in larger series of hemangiopericytic tumors.
In conclusion, DNA ploidy and SPF determined by highresolution DNA flow cytometry are useful indicators of different
biological behaviors in meningiomas. In tumors with diploid and
aneuploid DNA stem lines, the aneuploid line exhibited a significantly higher proliferative activity (SPF, PI).
120
ZELLNER ET AL.
REFERENCES
AHYAI, A. and SPAAR, F.W., DNA and prognosis of meningiomas: a
comparative cytological and fluorescence cytophotometrical study of 71
tumors. Acta Neurochir., 87, 119–141 (1987).
AKACHI, K., MATSUMOTO, M., YASUE, M., NAKAMURA, N., KAMADA, M. and
OHNO, T., DNA analysis of meningiomas using paraffin-embedded surgical
specimens in connection with clinical recurrence. No. Shinkei. Geka., 19,
1129–1134 (1991).
APPLEY, A.J., FITZGIBBONS, P.L., CHANDRASOMA, P.T., HINTON, D.R. and
APUZZO, M.L., Multiparameter flow cytometric analysis of neoplasms of
the central nervous system: correlation of nuclear antigen p105 and DNA
content with clinical behaviour. Neurosurgery, 20, 688–694 (1990).
ASSIETTI, R., BUTTI, G., MAGRASSI, L., DANOVA, M., RICCARDI, A. and
GAETANI, P., Cell-kinetic characteristics of human brain tumors. Oncology,
47, 344–351 (1990).
BUTTI, G., GAETANI, P., DANOVA, M., ASSIETTI, R., GIRINO, M. and
RICCARDI, A., Cell kinetic studies of human intracranial tumors. J.
neurosurg. Sci., 33, 47–53 (1989).
COON, J.S., LANDAY, A.L. and WEINSTEIN, R.S., Advances in flow cytometry
for diagnostic pathology. Lab. Invest., 57, 453 (1987).
COONS, S.W. and JOHNSON, P.C., Regional heterogeneity in the DNA
content of human gliomas. Cancer, 72, 3052–3060 (1993).
COONS, S.W., JOHNSON, P.C. and PEARL, D.K., Prognostic significance of
flow cytometry DNA analysis of human oligodendrogliomas. Neurosurgery, 34, 680–687 (1994).
CRONE, K.R., CHALLA, V.R., KUTE, T.E., MOODY, D.M. and KELLY, D.L.,
JR., Relationship between flow cytometric features and clinical behaviour of
meningiomas. Neurosurgery, 23, 720–724 (1988).
CRUZ-SANCHEZ, F. F., MIQUEL, R., ROSSI, M.L., FIGOLS, J., PALACIN, A. and
CORDESA, A., Clinico-pathological correlations in meningiomas: a DNA
and immunohistochemical study. Histol. Histopathol., 8, 1–8 (1993).
DEAN, P. and JETT, J., Mathematical analysis of DNA distributions derived
from flow microfluorimetry. Cytometry, 1, 71–77 (1980).
DRESSLER, L.G., Controls, standards and histogram interpretation in DNA
flow cytometry. Methods Cell Biol., 33, 157–171 (1990).
ENZINGER, F.M. and WEISS, S.W., Hemangiopericytoma. In: F.M. Enzinger
and S.W. Weiss (eds.), Soft tissue tumors, 2nd ed., pp. 596–613, C. Mosby,
St. Louis (1988).
FINN, W.G., GOOLSBAY, C.L. and RAO, S., DNA flow cytometric analysis of
hemangiopericytoma. Amer. J. clin. Pathol., 101, 181–185 (1994).
GIARETTI, W. and SANTI, L., Tumor progression by DNA flow cytometry in
human colorectal cancer. Int. J. Cancer, 45, 597–603 (1990).
HEIM, S. and MITELMAN, F., Cancer cytogenetics, pp. 243–245, A.R. Liss,
New York (1987).
HIDDEMANN, W., SCHUMANN, J. and ANDREEF, M., Convention on nomenclature for DNA cytometry. Cancer Genet. Cytogenet., 13, 181–183 (1984).
IRONSIDE, J.W., BATTERSBY, R.D.E., LAWRY, J., LOOMES, R.S., DAY, C.A.
and TIMPERLEY, W.R., DNA in meningioma tissues and explant cell
cultures: a flow cytometric study with clinicopathological correlates. J.
Neurosurg., 66, 588–594. (1987).
KATSUYAMA, J., PAPENHAUSEN, P.R., HERZ, F., GAZIVODA, P., HIRANO, A.
and KOSS, L.G., Chromosome abnormalities in meningiomas. Cancer
Genet. Cytogenet., 22, 63–68 (1986).
KAWAMOTO, K., NUMA, Y., FUJIWARA, H., OHUCHI, M. and MATSUMURA, H.,
Flow cytometric study on cell kinetics of brain tumors and their cultured
cells. Acta Neurochir., 97, 150–157 (1989).
KLEIHUES, R., BURGER, P.C. and SCHEITHAUER, B.W., Histological typing of
tumors of the central nervous system, Springer-Verlag, Berlin (1991).
KOBAYASHI, S. and HOSHINO, T., Clonogenicity and BCNU response of
subpopulations of human glioma sorted according to DNA content. No. To.
Shinkei, 37, 707–713 (1985).
LOOK, A.T., HAYES, F.A., NITSCHKE, R., MCWILLIAMS, N.B. and GREEN,
A.A., Cellular DNA content as a predictor of response to chemotherapy in
infants with unresectable neuroblastoma. N. Engl. J. Med., 311, 231 (1984).
MALTBY, E.L., IRONSIDE, J.W. and BATTERSBY, R.D.E., Cytogenetic studies
in 50 meningiomas. Cancer Genet. Cytogenet., 31, 199–210 (1988).
MATHIESEN, T., VON HOLST, H., ASKENSTEN, U. and COLLINS, P.V., DNA
determination in the clinical management of patients with meningioma or
hemangioblastoma. Brit. J. Neurosurg., 3, 575–582 (1989).
MAY, P.L., BROOME, J.C., LAWRY, J., BUXTON, R.A. and BATTERSBY, R.D.,
The prediction of recurrence in meningiomas. A flow cytometric study of
paraffin embedded archival material. J. Neurosurg., 71, 347–351 (1989).
MCCARTHY, R.C. and FETTERHOFF, T.J., Issues for quality assurance in
clinical flow cytometry. Arch. Pathol. Lab. Med., 113, 658–666 (1989).
MERKEL, D.E. and MCGUIRE, W.L., Ploidy, proliferative activity and
prognosis DNA flow cytometry of solid tumors. Cancer, 65, 1294–1205
(1990).
NAGASHIMA, T., HOSHINO, T., CHO, K.G., SENEGOR, M., WALDMAN, F. and
NOMURA, K., Comparison of bromodeoxyuridine labeling indices obtained
from tissue sections and flow cytometry of brain tumors. J. Neurosurg., 86,
388–392 (1988).
NISHIZAKI, T., OHSHITA, N. and NAGATSUGU, Y., Clinical evaluation of DNA
index in human brain tumors. J. Neurooncol., 17, 9–13 (1993).
NISHIZAKI, T., ORITA, T., KAJIWARA, K., IKEDA, N., OHSHITA, N., NAKYAMA,
H., FURUTAMI, Y., AKIMURA, T. and KAMIRYO, T., Correlation of in vitro
bromodeoxyuridine labeling index and DNA aneuploidy with survival or
recurrence in brain-tumor patients. J. Neurosurg., 73, 396–400 (1990).
OTTO, F., DAPI-staining of fixed cells for high-resolution flow cytometry of
nuclear DNA. Methods Cell Biol., 33, 105–110 (1990).
POULSGARD, L., RONNE, M. and SCHRODER, H.D., Cytogenetic studies of 19
meningiomas and their clinical significance. Anticancer Res., 9, 109–112
(1989).
QUIRKE, P., Flow cytometry: methodology and applications in pathology.
Arch. Pathol. Lab. Med., 113, 591–597 (1986).
RABINOVITCH, P.S., Handbook ‘‘Multicycle Vers. 2.5,’’ Phoenix Flow
Systems, Seattle.
RABSON, A.R., Flow cytometry in the diagnosis of brain tumors. Neurosurg.
Clin. N. Amer. 5, 135–146 (1994).
SANDBERG, A.A. and TURC-CAREL, C., The cytogenetics of solid tumors.
Relations to diagnosis, classification and pathology. Cancer, 59, 387–395
(1987).
SECKINGER, D., SUGARBAKER, E. and FRANKFURT, O., DNA content in
human cancer. Arch. Pathol. Lab. Med., 113, 619–626 (1989).
SHAPIRO, H.M., Flow cytometry of DNA content and other indicators of
proliferative activity. Arch. Pathol. Lab. Med., 113, 591–597 (1989).
SHAPIRO, J.R. and SHAPIRO, W.R., Clonal tumor cell heterogeneity. Prog.
exp. Tumor Res., 27, 49–66 (1984).
SPAAR, F.W., AHYAI, A. and BLECH, M., DNA-fluorescence cytometry and
prognosis (grading) of meningioma—a study of 104 surgically removed
tumors. Neurosurg. Rev., 10, 35–39 (1987).
WESTPHAL, M., HD´ ANSEL, M., KUNZMANN, R., HOLZEL, F. and HERRMANN,
H.D., Spectrum of karyotypic aberrations in cultured human meningiomas.
Cytogenet. Cell Genet., 52, 45–49 (1989).
WILLIAMS, D.L., TSIATIS, A., BRODEUR, G.M., LOOK, A.T., MELVIN, S.L.,
BOWMANN, W.P., KALWINSKY, D.K., RIVERA, G. and DAHL, G.V., Prognostic
importance of chromosome number in 136 untreated children with acute
lymphoblastic leukemia. Blood, 60, 864 (1982).
ZANG, K.D., Cytological and cytogenetical studies on human meningioma.
Cancer Genet. Cytogenet., 6, 249–274 (1981).
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