  , .

183: 188–194 (1997)

‘REVERTANT’ DCIS IN HUMAN AXILLARY BREAST

CARCINOMA METASTASES
 . *,  . ,  . ,  .    . 
Department of Pathology and Revlon/UCLA Breast Center, UCLA School of Medicine, Los Angeles, CA 90024, U.S.A.

SUMMARY
Recent experimental evidence obtained in Scid mice has suggested that the metastatic process is in large part epigenetically regulated
and undergoes partial reversion once the metastatic process is completed: the metastatic colonies become more engaged in the process
of growing in situ than actively metastasizing. Based on this experimental evidence, examples were sought of metastatic human cancers
where similar reversion to an in situ growth state was occurring. Review of 200 cases of metastatic human breast cancer revealed a 21
per cent incidence of reversion to a ductal carcinoma in situ (DCIS) growth pattern within axillary nodal metastases. The revertant
DCIS areas were characterized by an intact and circumferential basement membrane, as demonstrated by extracellular laminin and type
IV collagen immunoreactivity. These revertant DCIS areas could be distinguished from primary DCIS, however, by the absence of
surrounding myoepithelial cells in the former, identified in the latter by their positive maspin, S-100, and smooth muscle actin
immunoreactivity. The pattern of revertant DCIS, poorly differentiated (comedo) (13 per cent), intermediate (non-comedo) (6 per cent),
or well-differentiated (non-comedo) (2%), exhibited complete 100 per cent concordance with the primary DCIS pattern. The
concordance of histological patterns held true for even the subtypes of DCIS determined by architectural pattern, such as the
micropapillary or cribriform subtypes. Nuclear size by digital image analysis and Her-2/neu, p53, and Ki-67 status in the revertant DCIS
also exhibited complete concordance with the primary DCIS counterparts. Cases exhibiting a revertant DCIS pattern tended to be
ER-negative/EGFR-positive and exhibited significant nodal involvement (mean number, 9; mean area, 90 per cent) compared with cases
lacking a revertant pattern (mean number, 4; mean area, 15 per cent) (P<0·01) These findings suggest that reversion of the metastatic
phenotype may also be occurring within autochthonous human metastasis. ? 1997 John Wiley & Sons, Ltd.

J. Pathol. 183: 188–194, 1997.

No. of Figures 2. No. of Tables 0. No. of References 30.
KEY WORDS—DCIS; breast carcinoma; basement membrane; myoepithelial cells; autochthonous metastasis

INTRODUCTION our search, we observed a pattern of ductal carcinoma

Recent studies of our laboratory and others1–3 have
indicated that the metastatic process is in large part
epigenetically regulated and hence reversible. Our find-
ings with experimental metastasis in Scid mice indicated
that the metastatic colonies did not subsequently engage
in appreciable metastasis, but rather engaged in growing
in situ. Other studies with several spontaneously
metastasizing human cell lines in nude and Scid mice
demonstrated metastasis only when the cells were
injected orthotopically.2 Weiss first introduced the con-
cept of a ‘transient metastatic compartment’ to explain
how apparently weakly metastatic tumour cells could be
influenced by certain environmental factors which
increased their metastatic potential.3 Conversely, the
absence of these positive factors, or the presence of other
negative factors, could decrease or reverse metastatic
potential. These prior observations and experimental
studies prompted us to search for examples in human
pathological material which supported this concept. In
*Correspondence to: Sanford H. Barsky, MD, Department of
Pathology, UCLA School of Medicine, Los Angeles, CA 90024,
U.S.A. E-mail: sbarsky@ucla.edu
Contract grant sponsor: USPHS; contract grant numbers:
CA71195, CA40225, CA01351.
Contract grant sponsors: California Institute for Cancer Research
(CICR); Cancer Research Coordinating Committee (CRCC); Jonsson
Comprehensive Cancer Center (JCCC).
CCC 0022–3417/97/100188–07 $17.50
? 1997 John Wiley & Sons, Ltd.
in situ (DCIS) growth in axillary nodal metastases of
human breast cancer which suggested to us that the
metastatic process in human autochthonous metastasis
may also be reversible.
MATERIALS AND METHODS
We decided to conduct in-depth studies of the fre-
quency and histopathology of this pattern of DCIS
growth reversion and to investigate further its features
with digital image analysis and immunocytochemistry.
Computer retrieval
Two hundred cases of axillary node-positive breast
carcinoma seen during the past 5 years at the Revlon/
UCLA Breast Center and Center for the Health
Sciences, University of California at Los Angeles, Los
Angeles, CA, were retrieved and reviewed.
Anatomical pathology studies
Anatomical pathology studies included histopatho-
logical, digital image analysis, and immunocytochemical
studies of both the primary breast carcinoma and its
corresponding axillary metastases.
Received 7 October 1996
Revised 7 January 1997
Accepted 19 March 1997
 REVERTANT DCIS IN AUTOCHTHONOUS METASTASIS
Diagnostic criteria—Modern diagnostic criteria for
the diagnosis and classification of DCIS types and
subtypes as established by recent studies4,5 were
applied. Specifically, DCIS was classified into poorly
differentiated, intermediately differentiated, and well-
differentiated on the basis of cytonuclear differentiation.
The presence or absence of central necrosis was not used
as a basis of this classification but the term comedo was
applied to poorly differentiated cases and the term
non-comedo was applied to both intermediately and
well-differentiated cases. DCIS was also subclassified
on the basis of architectural differentiation into solid,
cribriform, and micropapillary subtypes.
Digital image analysis studies—Digital image analysis
studies were designed to compare quantitatively the
nuclear size of primary DCIS with the revertant DCIS
noted within the axillary metastases. This analysis was
conducted with a digital imaging system, which utilized
a Leitz Dialux microscope linked to a Vidicon camera,
an IBM PC with PCVision digitizer, and Microscience
software.6 Ten fields were counted of both primary
DCIS and revertant DCIS from each case. Only fully
intact DCIS nuclei were counted. The cross-sectional
nuclear area exhibited by the DCIS patterns was
expressed as mean&standard deviation and compared.
Digital image analysis was also used to determine both
the percentage of nodal area involved by metastasis
and the percentage of nodal metastasis manifesting a
revertant pattern.
Immunocytochemical studies—Immunocytochemical
studies were designed to embellish and expand upon the
previous histopathological studies. Laminin, type IV
collagen, S-100 protein, smooth muscle actin, maspin,
Her-2/neu, p53, Ki-67. Oestrogen receptor (ER), and
epidermal growth factor receptor (EGFR) were detected
by established methods of immunocytochemical
analysis.7–15 Monoclonal/polyclonal antibodies and
other immunohistochemical reagents were purchased
from the following sources: Ki-67 MIB 1 (AMAC);
p53 Ab2 and Her-2/neu Ab3 (Oncogene Science); ER
Ab (Abbot); EGFR Ab (CIBA-Corning); rabbit anti-
human maspin (Pharmingen), anti-cow S-100, and
anti-smooth muscle actin (DAKO); biotinylated goat
anti-mouse (Zymed); horseradish peroxidase-conjugated
streptavidin (Vector); and aminoethylcarbazole—AEC
(Zymed). For Ki-67, Her-2/neu, and p53 assays, slides
were processed for antigen retrieval by boiling in 10 m
citric acid solution for 30 min. For Ki-67 antigen,
formalin-fixed paraffin sections also underwent
microwave-processing.14 Sections were incubated with
primary antibody (1/100 Ki-67, 1/300 Her-2/neu, 1/20
ER, 1/100 p53, 1/10 EGFR, 1/50 laminin, 1/50 type IV
collagen, 1/1000 S-100, 1/2000 smooth muscle actin, and
1/500 maspin) as used in previous studies.7,15–17 Sheep
anti-mouse IgG and goat anti-rabbit IgG were used as
secondary antibodies at 1/200 and 1/25 dilutions,
respectively. Antigen binding sites were then revealed by
incubating with the peroxidase substrate AEC [or
peroxidase polymerizing diaminobenzidine (DAB)],
producing an insoluble brilliant red precipitate (or
? 1997 John Wiley & Sons, Ltd.
189
brownish-black staining) at sites of antigen presence.
Nuclei of cells containing ER protein, Ki-67, and p53
stained; surface membranes of cells containing either
increased Her-2/neu antigen or increased EGFR antigen
stained. For ER, tissues with greater than 10 per cent
nuclear staining (weak to strong staining intensity) were
considered positive. For p53, EGFR, and Her-2/neu
staining, either the cells were uniformly negative or
25–100 per cent staining occurred, the latter pattern of
which was considered positive. For Ki-67, the number of
positive nuclei was expressed as a percentage of the total
nuclei. For all of these antigens, cytoplasmic granular
staining was considered non-specific.
Statistical analysis
Standard tests of statistical significance were con-
ducted. These included chi-square, the Cochran–
Mantel–Haenszel modified chi-square, and a two-tailed
t-test.
RESULTS
Review of 200 cases of axillary node-positive breast
carcinoma revealed an overall 21 per cent incidence of a
reversion to a DCIS growth pattern within the axillary
metastases (Fig. 1A). Reversions of all the major types
of DCIS were observed in this study. For example,
patterns of revertant DCIS included poorly differenti-
ated (comedo) DCIS (13 per cent) (Fig. 1B), well-
differentiated (non-comedo) DCIS (6 per cent) (Figs 1C
and 1D), and intermediately differentiated (non-
comedo) DCIS (2 per cent) (Fig. 1E). There was 100 per
cent concordance between the types of primary and
revertant DCIS present within individual cases. This
strict fidelity of histopathological pattern even held for
the subtypes of DCIS based on architectural differentia-
tion, such as solid (Fig. 1C), cribriform (Fig. 1D), and
micropapillary (Fig. 1E). In some cases, the revertant
DCIS even displayed dystrophic calcifications (Fig. 1F)
identical to those present within the primary DCIS.
Areas of revertant DCIS were surrounded by an intact
and circumferential basement membrane demonstrated
by extracellular laminin and type IV collagen immuno-
reactivity (Fig. 1G). Basement membrane immunoreac-
tivity was completely absent in the adjacent non-DCIS
areas present within the lymph nodes. Areas of revertant
DCIS could be distinguished from primary DCIS, how-
ever, by the complete absence of a layer of myoepithelial
cells in the former (Fig. 1H) but the presence of either a
continuous or a focally discontinuous layer of myo-
epithelial cells in the latter (Fig. 1I). Myoepithelial cells
were identified by strong S-100, smooth muscle actin,
and maspin immunoreactivity. Maspin, a member of the
serpin family of proteinase inhibitors,17 was also
strongly immunodetectable in the myoepithelial cell
layer surrounding normal breast ducts and acini; maspin
immunoreactivity, however, was absent in normal breast
ductal and acinar cells and in both the primary and the
revertant DCIS epithelial cells, suggesting that maspin
may be a myoepithelial-specific marker.
  , . 183: 188–194 (1997)
190 S. H. BARSKY ET AL.
Not only did the revertant DCIS growth pattern
closely recapitulate the primary DCIS pattern by histo-
pathological criteria, but by digital image analysis, the
nuclear sizes exhibited by the two DCIS areas were
essentially identical (P>0·25) (Fig. 2A). Strong concord-
ance of Ki-67, Her-2/neu, and p53 status between the
revertant DCIS and the primary DCIS within individual
cases was also observed. When the individual patients
were grouped into primary, revertant, poorly differenti-
ated (comedo), intermediately, and well-differentiated
(non-comedo), no differences were observed in these
markers between the revertant and primary DCIS
(P>0·25), but significant differences were observed
between the comedo and non-comedo groups (P<0·05)
(Figs 2B and 2C). ER and EGFR status in the revertant
DCIS exhibited strong concordance with their primary
DCIS pattern (Fig. 2C). The groups exhibiting revertant
primary comedo DCIS showed a higher Ki-67 status
(Fig. 2B) and a greater presence of p53 and Her-2/neu
alterations (Fig. 2C) compared with the groups exhibit-
ing revertant and primary non-comedo DCIS (P<0·05).
Cases exhibiting revertant DCIS were largely
ER-negative and EGFR-positive (Fig. 2C), irrespective
of whether their pattern was revertant comedo or
non-comedo DCIS.
Cases exhibiting a revertant DCIS pattern exhibited
significant nodal involvement (mean number, 9; mean
area, 90 per cent), irrespective of whether the revertant
DCIS pattern was comedo or non-comedo, compared
with cases not exhibiting revertant DCIS (mean number,
4; mean area 15 per cent) (P<0·01) (Fig. 2D). In these
cases of revertant DCIS, the majority of the node was
replaced by metastatic carcinoma and the revertant
DCIS pattern was observed in 15–25 per cent of the
node. In many cases of revertant DCIS, the overall
appearance of the nodal metastases bore a strong resem-
blance to that of the primary tumour, which usually
showed extensive intraductal carcinoma (EIC) admixed
with invasive carcinoma. In all of the 200 cases, most of
which had readily apparent primary DCIS or EIC, in
the primary tumour either a continuous or a focally
discontinuous layer of myoepithelial cells was always
present around the islands of tumour cells showing
extracellular basement membrane staining. We never
observed extracellular basement membrane staining
around any tumour island exhibiting a complete absence
of myoepithelial cells in the primary tumour.
DISCUSSION
Breast cancer development in vivo includes not only a
derangement of growth, but also a progression to an
invasive and highly metastatic phenotype.18,19 The
molecular and biochemical mechanisms behind this
progression are not well defined, but for decades path-
ologists have felt that ductal carcinoma in situ (DCIS), a
preinvasive proliferative growth state present within the
primary ductal lobular units of the breast, progresses to
invasive and subsequently metastatic breast cancer.
Recently, this belief has been given credence by loss of
heterozygosity (LOH) studies observing that the same
? 1997 John Wiley & Sons, Ltd.
genetic locus at chromosome 11q13 is altered in both
microdissected in situ and invasive breast cancer from
the same patient.20 In retrospective studies, pathologists
have noted an approximately 25 per cent incidence of
progression of DCIS to invasive cancer over a 6 to
10-year period,21,22 but whether additional genetic
events in the DCIS cells or paracrine events governed
by the myoepithelial or other host cells influence this
progression is not known.23
The cardinal histopathological distinction between
DCIS of all types and invasive breast cancer is the
presence of a circumferential or near-circumferential
basement membrane and myoepithelial cell layer around
the ducts involved by DCIS.7,24 The myoepithelial cells
are thought to contribute largely to the synthesis of the
basement membrane and to exert other important para-
crine influences on both the overlying normal ductal
epithelium and the DCIS. Even when the DCIS involves
the adjacent lobules (so-called ‘cancerization’ of the
lobules), the surrounding basement membrane and
myoepithelial layer largely remain intact. In invasive
breast cancer, on the other hand, the invading breast
cancer cells penetrate this basement membrane and
myoepithelial layer. Although some carcinomas, such as
squamous cell carcinomas, can retain a surrounding
basement membrane during active invasion,25 invasive
breast carcinomas of all types, including even the most
differentiated tubular carcinomas, consistently lack a
surrounding basement membrane. The observation in
this study of a recapitulation of basement membrane
around the islands of metastatic breast carcinoma cells,
cells which lack surrounding myoepithelial cells, is com-
pelling evidence that the carcinoma cells have acquired
an ‘in situ’ phenotype. The strong correlation of his-
topathological features, digital image analysis nuclear
parameters, and molecular markers between the rever-
tant and primary DCIS areas of individual cases indi-
cates that this phenomenon is not stochastic but highly
directed.
These findings have value in our understanding of the
metastatic process in human breast cancer, because the
recognition of the revertant DCIS phenomenon sup-
ports the hypothesis that metastatic carcinomas can
recreate their primary microenvironment at a distant site
and engage in in situ growth rather than subsequent
metastasis. This hypothesis receives additional support
from many other observations that have been made in
diverse tumours, such as the presence of sustentacular
cells at the periphery of the ‘Zellballen’ of invasive and
metastatic paraganglioma, the occurrence of follicular
dendritic cells in the nodules of follicular lymphoma that
has spread outside the lymph node, and the presence of
non-neoplastic T-lymphocytes in between the tumour
cells of metastatic thymoma. In the case of metastatic
breast carcinoma, the attempt to recreate the micro-
environment of the primary tumour at a distant site
manifests itself as a reversion to a DCIS growth state.
Interestingly, the phenomenon that we are describing
is well recognized anecdotally among pathologists.
When we showed our cases to our pathology colleagues
at our institution, many responded that they had
‘seen these patterns before’. We recently learned that
  , . 183: 188–194 (1997)
 REVERTANT DCIS IN AUTOCHTHONOUS METASTASIS 191

Fig. 1—A–F
Fig. 1—(A) Revertant DCIS pattern (left) is seen juxtaposed to invasive areas (right) in lymph node metastases. Revertant DCIS patterns
included (B) poorly differentiated (comedo); (C) well-differentiated solid, (D) well-differentiated cribriform; (E) intermediately differentiated
micropapillary. There was strict fidelity between the type and subtype of primary DCIS and revertant DCIS. Some cases of nodal revertant
DCIS showed dystrophic luminal calcifications (F). All of the patterns of nodel revertant DCIS were surrounded by an intact basement
membrane, as evidenced by strong immunoreactivity to both type IV collagen and laminin (depicted) (G). Adjacent non-DCIS areas were
completely devoid of this extracellular immunoreactivity. The patterns of nodal revertant DCIS were, however, devoid of surrounding
myoepithelial cells (H), which were consistently present around primary DCIS of all types (I). Myoepithelial cells could be identified by
maspin (depicted), S-100 or smooth muscle actin positivity. (A, B, C, D, E, F) H & E, #150–#400; (G, H, I) Anti-laminin/anti-maspin,
immunoperoxidase, #250–#400

Professor N. F. Gowing, while working at the Royal
Marsden Hospital in the 1970s, made similar observa-
tions of a DCIS growth pattern in axillary lymph nodes
using PAS staining for basement membranes (personal
communication). Recently we published an experimental
study1 where we examined the issues of the paracrine
regulation of the metastatic process. In that study we
? 1997 John Wiley & Sons, Ltd.
observed that our experimental metastases did not
engage in subsequent metastasis, but rather engaged in
growing in situ. Based on these experimental studies, we
reasoned that a large component of the metastatic
process was epigenetically regulated and hence revers-
ible. Based on this earlier experimental study which we
conducted in Scid mice, we began looking for examples
  , . 183: 188–194 (1997)
192 S. H. BARSKY ET AL.
Fig. 1—G–I
in human pathological material that supported our
overall hypothesis. The present study is an observational
account of a DCIS pattern of growth in metastases of
human breast cancer which suggests that reversion to a
non-invasive in situ growth state may also be occurring
in human autochthonous metastasis. We cannot be
sure, however, that this growth pattern represents true
biological reversion.
Despite this, we believe that the areas of ‘revertant’
DCIS represent more than duct-like differentiation in an
invasive tumour. The proof of this argument lies in the
consistent presence of surrounding basement mem-
branes around these islands of revertant DCIS. Even the
most differentiated invasive ductal carcinomas, such as
tubular carcinomas, lack surrounding basement mem-
brane structures. Hence duct-like differentiation per se is
not sufficient to recapitulate basement membrane immu-
noreactivity. Furthermore, in the cases demonstrating
nodal revertant DCIS, analysis of the primary lesions
revealed no evidence of similar structures containing
basement membranes but lacking myoepithelial cells.
We are aware that some cases of primary breast cancer
might exhibit areas of a DCIS-like pattern which are
nevertheless invasive and lack surrounding myoepithe-
lial cells and basement membrane immunoreactivity.
This could explain the finite risk of nodal metastasis in
apparently ‘pure’ DCIS. This possibility was first sug-
gested in the 1960s by Professor Carlo Sirtori, who
argued in the Italian literature that intraductal carci-
noma of the breast could never be regarded with 100 per
? 1997 John Wiley & Sons, Ltd.
cent certainty as being a true carcinoma ‘in situ’.26
We did not observe in any of our 200 cases examples
of islands of invasive breast carcinoma lacking myo-
epithelial cells but exhibiting basement membrane
immunoreactivity in the primary tumour.
Another potential concern was whether the revertant
DCIS pattern that we were observing actually repre-
sented tumour cells lodged in basement membrane-
containing lymph node sinusoids. We considered this
alternative explanation for our observations, but
decided that it was unlikely for the following reasons:
firstly, we did not see membrane-bound structures
present in metastatic deposits outside nodes. Secondly,
in virtually all of the cases, the areas of revertant DCIS
were present in medullary and cortical-medullary areas
of the lymph node, sparing the subcortical lymphatic
spaces which were visible but not involved by metastatic
tumour. We felt it unlikely for metastatic deposits within
nodal lymphatics to involve selectively only medullary
nodal areas, sparing subcortical lymphatics. Thirdly, in
a substantial fraction of cases the nodes contained a
desmoplastic reaction and the pattern of revertant DCIS
was contained within these desmoplastic areas. When
metastases occupy nodal lymphatics or when there is a
sinus histiocytosis involving nodal lymphatics, there
usually is not a desmoplastic reaction.
Our studies indicated that many of the known
markers of breast carcinoma progression, Her-2/neu,
p53, loss of ER, increased EGFR expression, and
increased Ki-67, exhibited a strong correlation between
  , . 183: 188–194 (1997)
 REVERTANT DCIS IN AUTOCHTHONOUS METASTASIS 193

Fig. 2—(A) Nuclear size (mean&standard deviation) strongly correlated between the revertant and primary DCIS areas; poorly differentiated DCIS
(comedo) exhibited a larger nuclear size than intermediately and well-differentiated DCIS (non-comedo). (B ) Correlation of Ki-67 immunoreactivity
(mean percentage of positivity&standard deviation) was strong between revertant and primary DCIS for both poorly differentiated (comedo) and
intermediately and well-differentiated (non-comedo) DCIS, which, differed significantly from each other. (C) Correlation between revertant DCIS
(R-DCIS) and primary DCIS (P-DCIS) in the expression (percentage of cases where marker was positive) of different molecular markers implicated
in human breast carcinoma progression. (D) Nodal involvement: the number of nodes and the percentage of nodal area (mean&standard deviation)
were greater in cases in which a pattern of revertant DCIS was observed, suggesting that a metastatic size threshold is required to initiate the
revertant phenotype

the revertant and primary DCIS areas and hence altera-
tions in these markers were not the cause of the DCIS
revertant phenotype. As anticipated, both revertant and
primary poorly differentiated (comedo) DCIS exhibited
an increase in nuclear size, the Ki-67 proliferating
marker, and frequency of p53 and Her-2/neu alterations
compared with their intermediately and well-
differentiated (non-comedo) DCIS counterparts.27,28
Cases of revertant DCIS tended to be EGFR--positive
and ER-negative, irrespective of whether their pattern
was comedo or non-comedo; furthermore, cases of
revertant DCIS, irrespective of whether their pattern
was comedo or non-comedo, exhibited significant nodal
involvement in terms of both the number of nodes and
the percentage of nodal area involved. These observa-
tions indicated to us that the revertant phenomenon is a
function of the metastatic tumour volume/mass and is
? 1997 John Wiley & Sons, Ltd.
not usually observed when the metastatic tumour
volume/mass falls below a certain threshold. Cases
which are EGFR-positive/ER-negative represent a sub-
set of breast carcinoma cases which exhibit more
aggressive biological behaviour and more lymph node
metastases than cases which are EGFR-negative/
ER-positive.
Since no internal cellular (nuclear, cytoplasmic, or
membrane) markers exist for breast carcinoma which
reliably distinguish DCIS cells from invasive carcinoma
cells, we could only rely upon the extracellular basement
membrane markers, laminin and type IV collagen, to
distinguish invasive from DCIS (primary or revertant)
phenotypes. Recent in situ hybridization studies of
the calcium-binding protein psoriasin have observed
increased expression in the in situ versus the in-
vasive component of the same breast tumour.29 In
  , . 183: 188–194 (1997)
194 S. H. BARSKY ET AL.
collaborative studies we are investigating whether
increased psoriasin expression occurs in revertant DCIS.
Such a demonstration would provide additional proof of
a true reversion to a DCIS state.
Previous detailed pathological studies on the mor-
phology of axillary metastases of human breast cancer
have indicated strong morphological identity in terms of
glandular differentiation between the primary invasive
carcinoma and the lymph node metastases.30 These
studies have indicated that during human breast carci-
noma metastasis, selection for morphologically more
aggressive clones does not occur. Our present observa-
tions extend these findings. Not only do axillary meta-
stases exhibit morphological identity with their primary
invasive carcinoma counterparts, but high volume/mass
axillary metastases exhibit morphological identity with
their primary DCIS counterparts as well. This suggests
that axillary metastases are not the product of clonal or
genetic selection, but rather represent cellular popula-
tions epigenetically derived from the primary tumour,
capable of reverting back to their in situ growth state.
ACKNOWLEDGEMENTS
This work was supported by USPHS grants CA71195,
CA40225, and CA01351, and funds from the California
Institute for Cancer Research (CICR), the Cancer
Research Coordinating Committee (CRCC), and the
Jonsson Comprehensive Cancer Center (JCCC). Dr
Doberneck is a recipient of an American Cancer Society
Fellowship. Dr Barsky is a recipient of a NCI Research
Career Development Award.
REFERENCES
1. Safarians S, Sternlicht MD, Freiman CJ, Huaman JA, Barsky SH. The
primary tumor is the primary source of metastasis in a human melanoma/
Scid model. Implications for the direct autocrine and paracrine epigenetic
regulation of the metastatic process. Int J Cancer 1996; 66: 151–158.
2. Stephenson RA, Dinney CPN, Gohji K, Ordonez NG, Killion IJ, Fidler IJ.
Metastatic model for human prostate cancer using orthotropic implantation
in nude mice. J Natl Cancer Inst 1992; 84: 951–956.
3. Weiss L. Metastatic inefficiency. Adv Cancer Res 1990; 54: 159–211.
4. Holland R, Peterse JL, Millis RR, et al. Ductal carcinoma in situ: a proposal
for a new classification. Semin Diagn Pathol 1994; 11: 167–180.
5. Silverstein MJ, Poller DN, Waisman JR, et al. Prognostic classification of
breast ductal carcinoma-in-situ. Lancet 1995; 345: 1154–1157.
6. Fu YS, Cheng L, Huang I. DNA ploidy analysis of cervical condyloma and
intraepithelial neoplasia in specimens obtained by punch biopsy. Anal Quant
Cytol Histol 1989; 11: 187–195.
? 1997 John Wiley & Sons, Ltd.
7. Barsky SH, Siegal GP, Jannotta F, Liotta LA. Loss of basement membrane
components by invasive tumors but not by their benign counterparts. Lab
Invest 1983; 49: 140–148.
8. Cossman J, Schlegal R. p53 in the diagnosis of human neoplasia. J Natl
Cancer Inst 1991; 18: 980–981.
9. Visakorpi T, Kallioniemi OP, Koivula T, Harvey J, Isola J. Expression of
epidermal growth factor receptor and ERB2 (Her-2/neu) oncoprotein in
prostatic carcinomas. Mod Pathol 1992; 5: 643–648.
10. Cattoretti G, Becker MH, Key G, et al. Monoclonal antibodies against
recombinant parts of the Ki67 antigen (MIB 1 and MIB 3) detect prolifer-
ating cells in microwave-processed formalin-fixed paraffin sections. J Pathol
1992; 168: 357–363.
11. Allred D, Clark G, Molina R, et al. Overexpression of Her-2/neu and its
relationship with other prognostic factors change during the progression of
in situ to invasive breast cancer. Hum Pathol 1992; 23: 974–979.
12. Cheng L, Binder S, Fu Y, Lewin K. Demonstration of estrogen receptors by
monoclonal antibody in formalin-fixed breast tumors. Lab Invest 1988; 58:
346–353.
13. Greene G, Nolan C, Engler J, Jensen E. Monoclonal antibodies to human
estrogen receptor. Proc Natl Acad Sci USA 1990; 77: 5115–5119.
14. Shana-Rong S, Key ME, Kalra KL. Antigen retrieval in formalin-fixed
paraffin-embedded tissues: an enhancement method of immunohistochemi-
cal staining based on microwave oven heating of tissue sections. J Histo-
chem Cytochem 1991; 39: 741–748.
15. Barsky SH, Layfield L, Varki N, Bhuta S. Two human tumors with high
basement membrane-producing potential. Cancer 1988; 61: 1798–1806.
16. Sternlicht MD, Safarians S, Rivera SP, Barsky SH. Characterizations of the
extracellular matrix and proteinase inhibitor content of human myoepi-
thelial tumors. Lab Invest 1996; 74: 781–796.
17. Zou Z, Anisowicz A, Hendrix MJC, et al. Maspin, a serpin with tumor-
suppressing activity in human mammary epithelial cells. Science 1994; 263:
526–529.
18. Kohn EC, Liotta LA. Molecular insights into cancer invasion: strategies for
prevention and intervention. Cancer Res 1995; 55: 1856–1862.
19. Liotta L, Steeg P, Stetler-Stevenson W. Cancer metastasis and angiogenesis:
an imbalance of positive and negative regulation. Cell 1991; 64: 327–336.
20. Zhuang Z, Merino MJ, Chuaqui R, Liotta LA, Emmert-Buck MR. Identi-
cal allelic loss on chromosome 11q13 in microdissected in situ and invasive
human breast cancer. Cancer Res 1995; 55: 467–471.
21. Page D, Dupont W, Rogers L, Landenberger M. Intraductal carcinoma of
the breast: follow-up after biopsy only. Cancer 1982; 49: 751–758.
22. Rosen P, Braun D, Kinne D. The clinical significance of pre-invasive breast
carcinoma. Cancer 1980; 46: 919–925.
23. Sternlicht MD, Safarians S, Calcaterra TC, Barsky SH. Establishment and
characterization of a novel human myoepithelial cell line and matrix
producing xenograft from a parotid basal cell adenocarcinoma. In Vitro Cell
Dev Biol 1996; 32: 550–563.
24. Lagios MD, Westdahl PR,. Margolin FR, Rose MR. Duct carcinoma in
situ. Cancer 1982; 50: 1309–1314.
25. Gusterson BA, Warburton MJ, Mitchell D, Kraft N, Hancock WW.
Invading squamous cell carcinoma can retain a basal lamina. Lab Invest
1984; 51: 82–87.
26. Sirtori C, Talamazzi F. Il carcinoma intraduttale della mammella non e’mai
un carcinoma in situ. Tumori 1967; 53: 641–644.
27. Van de Vijver M, Peterse J, Mooi W, et al. Neu-protein overexpression in
breast cancer. N Engl J Med 1988; 319: 1239–1245.
28. Meyer JS. Cell kinetics of histologic variants of in situ breast carcinoma.
Breast Cancer Res Treat 1986; 7: 171–180.
29. Leygue E, Snell L, Hiller T, et al. Differential expression of psoriasin
messenger RNA between in situ and invasive human breast carcinoma.
Cancer Res 1996; 56: 4606–4609.
30. Sharkey FE, Greiner AS. Morphologic identity of primary tumor and
axillary metastases in breast carcinoma. Arch Pathol Lab Med 1985; 109:
256–259.
  , . 183: 188–194 (1997)