close

Вход

Забыли?

вход по аккаунту

?

478

код для вставкиСкачать
Publication of the International Union Against Cancer
Publication de l’Union Internationale Contre le Cancer
Int. J. Cancer: 70, 502–507 (1997)
r 1997 Wiley-Liss, Inc.
INTERPHASE KARYOTYPIC ANALYSIS OF CHROMOSOMES 11, 17 AND X
IN INVASIVE SQUAMOUS-CELL CARCINOMA OF THE CERVIX:
MORPHOLOGICAL CORRELATION WITH HPV INFECTION
Shirley A. SOUTHERN1 and C. Simon HERRINGTON1*
1Department of Pathology, University of Liverpool, Liverpool L69 3GA, UK
Human papillomavirus (HPV) infection has been widely
implicated in cervical carcinogenesis, but it appears to be an
early event, with other genetic abnormalities being required
for biological transformation. In this study, interphase cytogenetic analysis of numerical abnormalities of chromosomes 11,
17 and X was performed on paraffin-embedded tissue sections from 25 invasive squamous-cell carcinomas of the cervix
and compared with both histopathological features and the
morphological distribution of HPV sequences as determined
by in situ hybridisation. Numerical differences between chromosomes were identified in 76% of cases, with underrepresentation of chromosomes 11 and/or 17 relative to X in
64% of the total; 22 of 25 cases were HPV-positive, containing
either HPV 16, 18 or 31. There was no relationship between
the distribution of viral sequences and chromosomal pattern,
suggesting that HPV infection precedes karyotypic changes.
Our findings suggest that relative reduction in number of
chromosomes 11 and 17 is important in the development of
invasive cervical neoplasia and are consistent with the putative presence of relevant tumour-suppressor genes on these
chromosomes. Int. J. Cancer, 502–507, 1997.
r 1997 Wiley-Liss, Inc.
Human papillomavirus (HPV) infection has been widely implicated in cervical carcinogenesis (Herrington, 1994a). However,
current evidence suggests that HPV infection is an early event and
that other abnormalities, whether induced directly by HPV or
indirectly by other means, are required for biological transformation (Herrington, 1995). As the transformed phenotype is associated in vitro with genetic abnormalities and chromosomal abnormalities have been reported in cervical tumours (Teyssier, 1989), it is
logical to hypothesize that in vivo transformation may be associated with similar chromosomal changes.
Cytogenetic analysis of cervical tumours has shown that chromosomes 1, 3, 11 and 17 are commonly abnormal (Teyssier, 1989).
Induction of aneuploidy by HPV 16 sequences suggests that viral
DNA may play a part in this process and chromosomal abnormalities can be induced by HPV DNA in both cell (Matlashewski et al.,
1988) and raft (McCance et al., 1988) cultures.
Based on the study of somatic cell hybrids, it has been postulated
that cellular genes are capable of suppressing HPV gene expression
(zur Hausen, 1986). Although not directly identified, it has been
suggested that an element responsible for this suppression is
located on chromosome 11 (Bosch et al., 1990; Jesudasan et al.,
1995). Numerical abnormalities of chromosome 11 are demonstrable in cervical carcinoma-derived cell lines by interphase
cytogenetics (Herrington et al., 1995), and breakpoints in cervical
tumours tend to cluster at 11q23 (Sreekantaiah et al., 1991), a site at
which a high frequency of allele loss has been observed using
microdissection and minisatellite/PCR techniques (Bethwaite et
al., 1995). Abnormalities of chromosome 17 have also been
described in cytogenetic studies, and amplification of the c-erb-B2
gene has been demonstrated in a proportion of invasive cervical
tumours (Mitra et al., 1994). These observations support the
possibility that genes on chromosomes 11 and 17 are important in
cervical neoplasia.
One approach to the assessment of the genetic content of tissue
samples is the estimation of tumour ploidy by either flow or static
cytometry (Padberg et al., 1992; Takahashi et al., 1994). However,
these techniques are insensitive to minor changes in DNA content
and, in cervical neoplasia, are complicated by the presence of viral
DNA, which may significantly alter the overall DNA content of
infected cells, particularly in the presence of viral replication. An
alternative approach involves direct demonstration of karyotypic
abnormalities either by metaphase cytogenetics, in which chromosomes are dispersed in metaphase after limited culture, or interphase cytogenetics, in which intact cells are used (Herrington,
1994b). For the assessment of intact tissues, interphase cytogenetics is required, a further advantage being that this approach can be
applied to archival material, allowing retrospective study and the
correlation of molecular and morphological features.
At the time of writing, no published studies evaluating karyotypic abnormalities directly within intact cervical tissues were
identified. The aims of our study were, therefore, (i) to evaluate
numerical abnormalities of chromosomes 11, 17 and X in paraffin
sections from 25 invasive squamous cervical carcinomas, using the
X chromosome as a ‘‘control’’ in view of it being infrequently
abnormal in cytogenetic studies; (ii) to correlate the interphase
karyotype with morphological features; (iii) to investigate intratumoural heterogeneity of chromosome number; and (iv) to analyse
the relationship between karyotypic changes and the morphological
distribution of HPV infection as assessed by in situ hybridisation.
METHODS
Choice of cases
Twenty-five consecutive cases of invasive squamous cervical
carcinoma were selected from the diagnostic files of the Royal
Liverpool University Hospital. Slides were reviewed and tumours
classified according to the WHO classification of cervical tumours
(Scully et al., 1994). Grading was performed using the Broder
system (Kurman et al., 1992). Parallel 6 µm paraffin sections were
cut for interphase cytogenetic analysis and HPV typing by in situ
hybridisation.
Probes
The chromosome-specific probes used were biotinylated D11Z1,
D17Z1 and DXZ1 (Oncor, Gaithersburg, MD). These probes have
been localised previously to the peri-centromeric region of the
appropriate chromosome (Herrington et al., 1995). Individual HPV
plasmid clones for HPV 6, 11, 16, 18, 31 and 33 were labelled with
digoxigenin by nick translation as previously described (Herrington et al., 1989).
Interphase cytogenetics
The method used was based on that of Hopman et al. (1992).
Briefly, paraffin sections were dewaxed and pre-treated for 10 min
in 1 M NaSCN at 80°C. Sections were washed in water, then
digested with 0.4% (w/v) pepsin (Sigma P7012, Poole, UK) in 0.2
M HCl for 20–30 min at 37°C. Following washes in water, sections
were air-dried and the probe was applied at a concentration of 1
ng/µl in 2 3 SSC, 10% dextran sulphate, 60% formamide and 1
*Correspondence to: Department of Pathology, University of Liverpool,
Duncan Building, Royal Liverpool University Hospital, Liverpool L69
3GA, UK. Fax: 44-151-706-5859.
Received 21 August 1996; revised 5 November 1996.
CHROMOSOMAL ABNORMALITIES IN CERVICAL CARCINOMA
µg/ml sheared salmon sperm DNA. Denaturation was carried out at
80°C for 8 min and hybridisation at 37°C overnight. After
hybridisation, sections were washed with 60% formamide, 2 3
SSC, pre-equilibrated to pH 7.0 at 42°C for 20 min and then PBS
containing 3% (w/v) BSA and 0.05% Tween 20 (buffer A) for 20
min at room temperature. Detection was performed by sequential
incubation at 37°C in monoclonal anti-biotin (1:100), peroxidaseconjugated rabbit anti-mouse (1:80) and peroxidase-conjugated
swine anti-rabbit (1:100) for 30 min each. Washes were carried out
in buffer A. Signals were developed using diaminobenzidine
(DAB)/H2O2, and sections were counter-stained in haematoxylin
and mounted in DPX.
Three areas were marked on each slide, with care being taken
that the same tumour foci were analysed for each chromosome. In
each tumour, the foci analysed were chosen such that there was no
identifiable morphological difference between the areas, to minimise the possible effects of variation due to nuclear truncation. The
number of signals per nucleus was counted at a total magnification
of 3630, and the following rules were observed: (i) all signals were
counted; (ii) overlapping nuclei were not counted; (iii) split signals,
as defined previously (Herrington et al., 1995), were counted as
single signals. Signal number was recorded for each nucleus
individually in the order of counting. Tumour cell nuclei (n 5 200)
from each of the 3 separate areas were analysed from each tumour,
using each probe and frequency distribution generated. We have
shown previously that there is no significant intra- or inter-observer
variation using this approach (Southern and Herrington, 1996).
Signal distributions were generated from the same sections in the
same way from 100 stromal cells, using each probe in each case to
assess technical comparability.
503
FIGURE 2 – (a): Distributions obtained with chromosomes 11 and 17
from one area of case 18. ‘‘Trisomic’’ patterns are present with both
probes, but there is a significant difference between the distributions.
Plotting signal number as a function of counting order demonstrated a
trisomic population only for chromosome 17 but morphologically
discrete disomic and trisomic populations for chromosome 11 (b). This
case showed intra-tumoural heterogeneity with all 3 probes (see
Table I).
Distributions were compared using the 2-tailed Mann-Whitney
U test corrected for tied values (Altman, 1991). Heterogeneity was
assessed by statistical comparison of distributions obtained from
each area with the same probe and from the same area with each
different probe. Statistical analyses were corrected for multiple
comparisons using the Bonferroni method to avoid type I errors
(Altman, 1991): each distribution was compared with 4 other data
sets; therefore, the significance level used was p , 0.012.
HPV in situ hybridisation
Sections were dewaxed and digested using proteinase K at a
concentration of 0.5 mg/ml in 0.1 M Tris-HCl (pH 7.6), 0.1 M NaCl
(TBS) for 15 min at 37°C. Following washing in TBS, slides were
air-dried, the probe was applied and sections were denatured at
95°C for 6 min followed by hybridisation at 37°C overnight.
FIGURE 1 – Distributions generated from a single area of (a) case 15
showing monosomy 11 with no significant population of disomic cells
compared with disomy X and trisomy 17 and (b) a population
monosomic for chromosome 17 with disomy X and 11 in case 22. Note
in (b) that a significant disomic population is also present with
chromosome 17.
FIGURE 3 – Heterogeneity of chromosome number in case 16. There
was no significant difference between the areas analysed, but there is a
difference between the chromosomal distributions.
504
SOUTHERN AND HERRINGTON
FIGURE 4 – Interphase cytogenetic analysis of case 14 with probes for chromosomes 11 (a), 17 (b) and X (c). Frequency distributions generated
from these sections showed trisomic, disomic and tetrasomic populations, respectively (see Table I). HPV 18 sequences were widely distributed
throughout the tumour (d ). Note that the degree of nuclear positivity for HPV is not related to nuclear size. Scale bars 5 5 µm for a–c and 10 µm
for d.
CHROMOSOMAL ABNORMALITIES IN CERVICAL CARCINOMA
505
Cocktails of HPV 6, 11, 16, 18, 31 and 33 were used to screen for
HPV, and those cases positive were typed using individual HPV
probes at a concentration of 2 ng/µl in 50% formamide, 2 3 SSC,
10% dextran sulphate. Post-hybridisation washes were carried out
in 4 3 SSC followed by the following detection sequence:
monoclonal antidigoxin (1:5,000), biotinylated rabbit anti-mouse
antibody (1:200) and strepatividin alkaline phosphate conjugate
(1:100). The signal was developed with naphthol AS-MX phosphate/
Fast red. Control HPV-infected biopsies and cell lines (HeLa and
SiHa) were analysed in each experiment.
HPV positivity, type and signal distribution were determined
independently of the interphase cytogenetic analysis. Positivity
was classified as punctate or diffuse, as previously described
(Cooper et al., 1991). These patterns have been shown to correlate
with integrated and episomal viral sequences, respectively (Cooper
et al., 1991; Berumen et al., 1995).
RESULTS
Morphological analysis
Five of the 25 tumours were keratinising and 20 non-keratinising
squamous-cell carcinomas. No adenocarcinomas or small cell
carcinomas were included. Two tumours were grade 1, 18 grade 2
and 5 grade 3.
Interphase cytogenetics
Absolute chromosome number. The absolute chromosome number present in each tumour cannot be determined directly as the
effects of truncation of nuclei of variable sizes leads to distributions
which cannot be directly extrapolated to the karyotype of whole
cells. However, the significant population with the highest signal
number was determined previously using data derived from normal
epithelium (Southern and Herrington, 1996) to give an estimate of
chromosome number, in keeping with data reported by others
(Hopman et al., 1991).
The approach taken here was to ensure technical comparability
of results obtained with each probe by analysis of internal control
stromal cells. No significant difference was found between distributions obtained using each of the 3 probes analysed in any case,
allowing direct comparison of distributions obtained from the same
tumour area with different probes. Monosomic populations were
identified when there was a significant ‘‘left shift’’ of distributions
compared with other probes showing a disomic pattern in the same
tumour area (Fig. 1); this approach controls for the effects of
nuclear truncation and technical variation. Trisomic and tetrasomic
populations were identified by the presence of 3 or 4 signals,
respectively, in greater than 10% of nuclei; this cut-off is based on
sensitivity calculations derived previously using normal epithelium
(Southern and Herrington, 1996) and agrees with the cut-off used
by others (Wolman et al., 1992; Micale et al., 1994). Although each
tumour area is assigned a single chromosome number in this way,
this does not exclude the presence of other populations and does
not determine the proportion of the tumour that the population
constitutes. Thus, areas with the same assigned chromosome
number can have significantly different distributions due to the
presence of more than one population within that area (Fig. 2).
Nevertheless, this method gives a qualitative assessment of chromosome number.
Relative chromosome number and intra-tumoural heterogeneity.
As no significant difference was found between distributions
obtained from internal control stromal cells, distributions generated
from the same tumour area with different probes could be
compared statistically. Significant differences were recorded as
indicating relative under- or over-representation of the chromosomes (Figs. 3, 4). Intra-tumoural heterogeneity for individual
chromosomes was defined by the finding of a statistically significant difference between distributions obtained from different
tumour areas (Fig. 5).
FIGURE 5 – Distributions generated from case 8 using probes for
chromosomes 11 and X. Note the intra-tumoural heterogeneity, with a
mixture of disomic and tetrasomic populations. There is no significant
difference between the chromosomes.
The interphase cytogenetic results obtained with the 25 tumours
are shown in Table I. Briefly, 6 showed no difference between the
chromosomes: 4 were karyotypically normal with the 3 probes
used and 2 showed evidence of focal tetraploidisation. In 14 cases,
there was under-representation of chromosomes 11 and 17 relative
to the X chromosome (Figs. 3, 4): in 10 cases, there was
significantly greater loss of chromosome 17 than 11, in 2 there was
greater loss of chromosome 11 and in 2 there was equivalent loss of
chromosomes 11 and 17. In 2 cases, there was under-representation
of chromosome 17 relative to both chromosomes 11 and X. Three
cases showed under-representation of the X chromosome relative
to chromosomes 11 and 17.
Heterogeneity between tumour areas with the same chromosome
probe was present in 14 tumours. In only 4 cases was there
demonstrable heterogeneity with all 3 probes. Two of these cases
showed changes consistent with tetraploidisation, there being
tetrasomic populations present with all 3 probes, with no significant
difference between the distributions obtained with each probe (Fig.
5). In both of these cases, the tumour foci showing tetraploidisation
were morphologically localised but showed no morphological
differences.
HPV analysis
Of 25 tumours, 22 (88%) were HPV-positive by in situ hybridisation. All positive cases contained a punctate signal within tumour
cell nuclei, with 8 cases containing diffusely labelled nuclei in
addition (Fig. 3d); 19 of these cases were HPV 16-positive, 2 HPV
SOUTHERN AND HERRINGTON
506
TABLE I – MORPHOLOGICAL, INTERPHASE CYTOGENETIC AND HPV DATA1
Case
Morphology
11
17
X
Relative number
HPV type
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
NK G2
NK G2
K G1
NK G2
NK G2
NK G2
NK G3
NK G2
NK G2
NK G2
NK G2
K G2
NK G3
NK G2
NK G2
K G3
NK G3
NK G3
NK G2
NK G2
NK G2
K G1
K G2
NK G2
NK G2
2/3H
3
2
2
2
2
4
2/4H
4H
4
2
2
4H
3
1
3
3
3H
2
2/3H
2/3H
2
2
2/4H
2
2
2
2
2
2
1/2H
3
2/4H
2/4H
3H
2
2
4
2
3
2
2
3H
1/2
2/3H
2
1/2
2
2/4H
2
2/3H
3/4H
2
4
2
2/4H
4
2/4H
1/2/4H
4H
1/2H
2
4
4
2/4H
4
3
4H
3
3
2/3/4H
2
2
2/4H
1/2H
17 , 11 5 X
17 , 11 , X
11 5 17 5 X
11 5 17 , X
11 5 17 5 X
17 , 11 , X
17 , 11 , X
11 5 17 5 X
X , 17 , 11
17 , 11 , X
X , 11 5 17
11 5 17 5 X
17 , 11 , X
17 , 11 , X
11 , 17 , X
17 , 11 , X
17 , 11 , X
11 , 17 , X
17 , 11 , X
11 5 17 , X
17 , 11 , X
17 , 11 5 X
11 5 17 5 X
11 5 17 5 X
X , 11 5 17
16
16
16
16
162
—
16
162
162
—
16
16
312
182
16
18
16
16
162
16
16
162
16
—
162
1NK 5 non-keratinising; K 5 keratinising; G 5 tumour grade. Numbers refer to the karyotype identified
with probes for chromosomes 11, 17 and X. H denotes heterogeneity between separate areas. Relative
number refers to relative differences between signal distributions: , represent a significantly lower signal
number and 5 represents no significant difference between tumour distributions.–2Cases containing
diffusely positive cells by HPV in situ hybridisation.
18-positive and 1 HPV 31-positive. HPV 6, 11, and 33 were not
identified in any case. There was no relationship with chromosomal
pattern except in case 15, where single punctate signals were
present throughout the tumour but were markedly larger in the area
showing tetrasomy X. It is of note that this tumour had a unique
chromosomal pattern (Fig. 1a; Table I).
DISCUSSION
Our data demonstrate non-random numerical chromosome
changes in a series of invasive squamous-cell carcinomas of the
cervix, with relative under-representation of chromosome 11
and/or 17 compared with the X chromosome in 64% of the tumours
investigated. This finding is consistent with the hypothesis that
tumour-suppressor genes of importance in cervical carcinogenesis
are located on these chromosomes. More frequent and extensive
under-representation of chromosome 17 appears at first sight
discordant with the finding of gene amplification (e.g., of the
c-erb-B2 gene on chromosome 17) in some previous studies (Mitra
et al., 1994). However, an absolute numerical increase in chromosomes 11 and 17 was identified in 14 (56%) and 9 (36%) tumours,
respectively, in this series. This suggests that, in some cases, gene
amplification may in part be due to an increase in chromosome
number rather than linear amplification and that, although the
absolute number of a particular chromosome may be increased, its
relative number may be reduced. This analysis underscores the
necessity of using more than one chromosome probe to allow
assessment of relative chromosome number in interphase cytogenetic studies.
Overall, heterogeneity between chromosomes was detectable in
76% of tumours. Moreover, tetraploidisation was detectable when
morphologically localised. This process is thought to be associated
with tumour progression (Hopman et al., 1992) and may underlie
the induction of aneuploidy by HPV sequences. This is in keeping
with the presence of a significant population tetrasomic for
chromosome X in 10 of the 14 cases in which under-representation
of chromosomes 11 and 17 was identified. The 4 other cases
showed focal or widespread trisomy for one or more chromosomes,
suggesting that gain of individual chromosomes also occurs. A
further group of cases disomic for all 3 chromosomes or in which
there was focal loss of either chromosome 17 or, more commonly,
the X chromosome was identified. These findings suggest that
pathways other than tetraploidisation are important in squamous
carcinogenesis in some cases.
Three of the 5 keratinising tumours analysed were disomic for all
3 chromosomes compared with only one of the remaining 20
non-keratinising tumours. The fourth keratinising tumour showed
only focal monosomy 17, and the final case was HPV 18-infected
and of high nuclear grade. The lack of marked abnormality in most
of the keratinising tumours may partly reflect the low grade of these
lesions but also implies differences in the interaction between HPV
sequences and neoplastic squamous cells in the presence of
keratinisation.
Intra-tumoural heterogeneity with individual probes was identified in this study in 14 of 21 non-disomic tumours (67%). The
identification of heterogeneity is unlikely simply to reflect variation
in nuclear size between areas as only 2 of 5 grade 3 tumours, all of
which showed marked nuclear pleomorphism, demonstrated heterogeneity using this approach. It is of note that both of the HPV
18-positive tumours showed the same chromosomal pattern, with
no significant heterogeneity between tumour areas with any of the 3
probes analysed. This raises the possibility that HPV 18 infection is
associated with specific chromosomal changes; investigation of
this hypothesis requires analysis of a larger group of cases.
No relationship was identified between either the presence of
HPV sequences in these tumours, or the pattern of signal obtained,
and chromosome number, except in one case where focal increase
in signal size correlated with the presence of a tetrasomic X
chromosome population. This tumour had a chromosomal pattern
unique in this series. This observation is not likely simply to be due
to viral integration into the X chromosome as the variation
identified was in signal size, not number, suggesting that viral
amplification accompanied X chromosome duplication. Analysis of
the distribution of diffusely labelled cells in the 8 cases in which
CHROMOSOMAL ABNORMALITIES IN CERVICAL CARCINOMA
they were present showed no relationship with chromosomal
pattern. This was most clearly demonstrated by case 8, in which
separate islands of tumour showed tetraploidisation but no difference in either morphological appearance or HPV staining distribution. This suggests that tetraploidisation is not directly related to
viral DNA replication in these tumours. However, it does not
exclude the possibility that it is related to viral gene transcription as
all of the tumours contained a punctate viral signal, suggesting that
integrated viral sequences were also present (Cooper et al., 1991;
Berumen et al., 1995); these could equally lead to enhanced gene
transcription and, hence, induction of aneuploidy.
This study demonstrates that intra-tumoural karyotypic heterogeneity is frequent in invasive squamous carcinoma of the cervix. The
presence of HPV sequences, in a pattern which correlates with viral
integration, in the majority of cases suggests that HPV infection
507
precedes karyotypic changes and is consistent with the in vitro
demonstration that HPV infection can induce aneuploidy. The
relative chromosome number found in these cases suggests that
relative loss of chromosomes 11 and 17 is important in the
development of invasive cervical neoplasia. The identification of a
smaller group of tumours with disomy or isolated loss of the X
chromosome suggests that alternative pathways occur. Analysis of
intra-epithelial lesions should help to refine the point at which these
changes occur.
ACKNOWLEDGEMENTS
We thank the University of Liverpool and Wellbeing/RCOG for
funding.
REFERENCES
ALTMAN, D.G., Practical statistics for medical research, Chapman and
Hall, London (1991).
BERUMEN, J., UNGER, E.R., CASAS, L. and FIGUEROA, P., Amplification of
human papillomavirus type-16 and type-18 in invasive cervical cancer.
Hum. Pathol., 26, 676–681 (1995).
BETHWAITE, P., KORETH, J., HERRINGTON, C.S. and MCGEE, J.O.D., Loss of
heterozygosity occurs at the D11S29 locus on chromosome 11q23 in
invasive cervical cancer. Brit. J. Cancer, 71, 814–818 (1995).
BOSCH, F.X., SCHWARZ, E., BOUKAMP, P., FUSENIG, N.E., BARTSCH, D. and
ZUR HAUSEN, H., Suppression in vivo of human papillomavirus type 18
E6-E7 gene expression in nontumorigenic HeLa 3 fibroblast hybrid cells.
J. Virol., 64, 4743–4754 (1990).
COOPER, K., HERRINGTON, C.S., STICKLAND, J.E., EVANS, M.F. and MCGEE,
J.O.D., Episomal and integrated HPV in cervical neoplasia demonstrated by
nonisotopic in situ hybridization. J. clin. Pathol. 44, 990–996 (1991).
HERRINGTON, C.S., Human papillomaviruses and cervical neoplasia I:
virology, classification, pathology and epidemiology. J. clin. Pathol., 47,
1066–1072 (1994a).
HERRINGTON, C.S., Interphase cytogenetics: principles and applications. J.
Histotechnol., 17, 219–234 (1994b).
HERRINGTON, C.S., Human papillomaviruses and cervical neoplasia II:
interaction with other factors. J. clin. Pathol., 48, 1–6 (1995).
HERRINGTON, C.S., BURNS, J., GRAHAM, A.K., BHATT, B. and MCGEE,
J.O.D., Interphase cytogenetics using biotin and digoxigenin labelled
probes I: relative sensitivity of both reporter molecules for HPV16
detection in CaSki cells. J. clin. Pathol., 42, 592–600 (1989).
HERRINGTON, C.S., COOPER, K. and MCGEE, J.O.D., Interphase cytogenetics: analysis of numerical chromosome aberrations in isolated cells. J.
Pathol., 175, 283–295 (1995).
HOPMAN, A.H., VAN HOOREN, E., VAN DE KAA, C., VOOIJS, P.G. and
RAMAEKERS, F.C.S., Detection of numerical chromosome aberrations using
in situ hybridization in paraffin sections of routinely processed bladder
cancers. Mod. Pathol., 4, 503–513 (1991).
HOPMAN, A.H.M., PODDIGHE, P., MOESKER, O. and RAMAEKERS, F.C.S.,
Interphase cytogenetics: an approach to the detection of genetic aberrations
in tumours. In: C.S. Herrington and J.O’D. McGee (eds.), Diagnostic
molecular pathology: a practical approach, vol. 1, pp. 141–167, Oxford
University Press, Oxford (1992).
JESUDASAN, R.A., RAHMAN, R.A., CHANDRASHEKHARAPPA, S., EVANS, G.A.
and SRIVATSAN, E.S., Deletion and translocation of chromosome 11q23
sequences in cervical carcinoma cell lines. Amer. J. hum. Genet., 56,
705–715 (1995).
KURMAN, R.J., NORRIS, H.J. and WILKINSON, E., Tumors of the cervix,
vagina and vulva. Atlas of tumor pathology (3rd series, Fasc. 4), Armed
Forces Institute of Pathology (1992).
MATLASHEWSKI, G., OSBORN, K., BANKS, L., STANLEY, M. and CRAWFORD,
L., Transformation of primary human fibroblast cells with human papillomavirus type 16 DNA and EJ-ras. Int. J. Cancer, 42, 232–238 (1988).
MCCANCE, D.J., KOPAN, R., FUCHS, E. and LAIMINS, L.A., Human papillomavirus type 16 alters human epithelial cell differentiation in vitro. Proc.
nat. Acad. Sci. (Wash.), 85, 7168–7173 (1988).
MICALE, M.A., VISSCHER, W., GULINO, S.E. and WOLMAN, S.R., Chromosomal aneuploidy in proliferative breast disease. Hum. Pathol., 25, 29–35
(1994).
MITRA, A.B., MURTY, V.V.V.S., PRATAP, M., SODHANI, P. and CHAGANTI,
R.S.K., Erb-B2 (Her2/neu) oncogene is frequently amplified in squamous
cell carcinoma of the uterine cervix. Cancer Res., 54, 637–639 (1994).
PADBERG, B.C., ARPS, H., FRANKE, U., THIEDEMANN, C., REHPENNING, W.,
STEGNER, H.E., LIETZ, H., SCHRODER, S. and DIETEL, M., DNA cytophotometry and prognosis in ovarian tumours of borderline malignancy—a
clinicomorphological study of 80 cases. Cancer, 69, 2510–2514 (1992).
SCULLY, R.E., BONFIGLIO, T.A., KURMAN, R.J., SILVERBERG, S.G. and
WILKINSON, E.J., Histological typing of female genital tract tumours. World
Health Organisation International Histological Classification of Tumours,
Springer-Verlag, Berlin (1994).
SOUTHERN, S.A. and HERRINGTON, C.S., The assessment of intra-tumoural
karyotypic heterogeneity by interphase cytogenetics in paraffin wax sections. J. clin. Pathol. mol. Pathol., 49, M283–M289 (1996).
SREEKANTAIAH, C., DE BRAEKELEER, M. and HAAS, O., Cytogenetic findings
in cervical carcinoma: a statistical approach. Cancer Genet. Cytogenet., 53,
75–81 (1991).
TAKAHASHI, Y., TAKENAKA, A., ISHIGURO, T. and NODA, Y., Intratumoral
DNA heterogeneity correlated with lymph node involvement and surgical
staging in epithelial ovarian cancer by flow cytometry. Cancer, 73,
3011–3014 (1994).
TEYSSIER, J.R., The chromosomal analysis of human solid tumours: a triple
challenge. Cancer Genet. Cytogenet., 37, 103–125 (1989).
WOLMAN, S.R., MACOSKA, J.A., MICALE, M.A. and SAKR, W.A., An
approach to definition of genetic alterations in prostate cancer. Diagn. mol.
Pathol., 1, 192–199 (1992).
ZUR HAUSEN, H., Intracellular surveillance of persisting viral infections:
human genital cancer results from deficient cellular control of papillomavirus gene expression. Lancet, ii, 489–491 (1986).
Документ
Категория
Без категории
Просмотров
2
Размер файла
523 Кб
Теги
478
1/--страниц
Пожаловаться на содержимое документа