2014, Volume 30, Number 1, Page(s) 018-022
Are Mammaglobin and Gcdfp-15 Sensitive Markers for Diagnosis of Metastatic Basal-Like Triple Negative Breast Carcinomas?
Nasser RAKHSHANİ, Arash DARYAKAR
Department of Pathology, Tehran University of Medical Sciences, Firuzgar Hospital, TEHRAN, IRAN ISLAMIC REPUBLIC
Keywords: Mammaglobin, Breast cancer, Immunohistochemistry, Differential diagnosis
There are small studies on expression of mammaglobin
and GCDFP-15 for detection of basal-like triple negative breast
cancer. This type of cancer has shorter survival and higher mortality
rate. There must be reliable markers for detecting this type of tumor
especially in metastatic cases with unknown origin.
Material and Method: In this study we assessed 66 paraffin
blocks of breast cancers previously diagnosed as triple negative
subtype in Mehr hospital (Tehran, Iran) by tissue microarray and
immunohistochemistry technique for expression of mammaglobin
Results: GCDFP-15 was positive in 12 cases (18.2%) and the other
54 (81.8%) cases remained negative. Mammaglobin was positive only
in 6 cases (9.1%) and the remaining 60 (90.9%) cases were negative.
Conclusion: According to recent studies and our findings, there is no
useful immunohistochemical marker for detection of breast source in
cases of metastatic triple negative breast cancer with unknown origin
and we must try hard to discover more accurate immunohistochemical
markers for these highly metastatic breast cancers.
Breast cancers are believed to be among the most common
cancers in women, and although diagnosis and treatment
techniques have improved a lot, they are still considered
as one of the main causes of death from cancers1
analysis of breast carcinoma has shown that there are
distinct subtypes of breast carcinoma that were first found
by Perou and his colleagues in 20002
Recently the subtypes of breast cancers recognized by their
gene signature include luminal (type A and B, and type
C), HER2/neu type, basal-like, and normal breast-like.
Attempts have been made to use immunohistochemistry
(such as a panel including antibodies to estrogen receptor
(ER), progesterone receptor (PR), HER2/neu, cytokeratin
5/6, epidermal growth factor receptor (EGFR) and Ki-
67 to assign tumors to various molecular subtypes3,4. The “gold standard” to determine molecular invasive
breast carcinoma (IBC) subtypes is gene expression array analysis, but this method requires fresh tissue. Surrogate
immunohistochemical profiles correlating to the molecular
subtypes have been developed5.
Triple negative breast cancer (TNBC) is a subgroup of
breast cancers that has been defined as negativity for ER, PR
and Her2/neu markers by immunohistochemistry (IHC). It
is also divided into 2 subgroups based on the expression of
basal markers. These subgroups that are named basal-like
and non-basal-like are positive and negative respectively
for myoepithelial/basal markers such as cytokeratin 5/6,
smooth muscle actin and EGFR6.
A shorter survival course and higher metastatic rate are
the characteristics of this type of breast cancer7. Reliable
markers should be found to identify this type of tumor
especially in metastatic cases whose origins are unknown.
The basal-like subtype is characterized by the expression
of myoepithelial / basal markers and molecular changes
including TP53 gene mutations, BRCA1 inactivation, and
many chromosomal alterations8-10.
Gross cystic disease fluid protein 15 (GCDFP-15) was
identified by Haagensen et al in 197711. GCDFP is
a 15-kDa monomer found in chromosome 7 and cells
with apocrine differentiation express it12. Normal
breast ducts and lobules do not represent GCDFP,
although apocrine metaplastic epithelium expresses it.
An immunohistochemical study performed by Wick
and colleagues in 1996, GCDFP-15 had a sensitivity and
specificity up to 74% and 95%, respectively, in primary
mammary carcinoma13. In another study by Fiel MI
et al, GCDFP-15 was found to be positive in up to 57%
of primary as well as metastatic breast carcinomas14.
Additionally Bhargava et al reported GCDFP-15 positivity
in just about 25% of breast carcinomas15.
Gene sequence of mammaglobin (MGB) was first discovered
in breast carcinoma tissue in 1994 by Watson and Fleming16. In 1996, they found complete DNA clone of MGB17.
In their study, MGB was positive in 50% of primary breast
carcinomas and 62% of metastatic breast carcinomas. In the
study of Han et al, sensitivity of mammaglobin antibody for
the detection of breast cancer was 84.3% when compared
with GCDFP-15 (44.3%) in a series of 70 breast cancer
cases18.This marker is also imparted in benign breast
epithelium. In this study we assessed paraffin blocks of
breast cancers previously diagnosed as basal-like TNBC in
Mehr Hospital (Tehran, Iran) using IHC technique for the
expression of mammaglobin and GCDFP-15 markers.
At the time of writing this article only study of Lewis et
al accessed sensitivity of mammaglobin and GCDFP-15 for detecting basal-like TNBC’s that were 24% and 5%,
respectively19. In attention to this fact that breast
carcinoma is one of the most common metastatic cancers
in women and TNBC’s are highly metastatic, we want
to assess sensitivity of GCDFP-15 and mammaglobin
markers for diagnosis of these cancers. As in the work
up of metastatic tumors with unknown origin when we
consider a breast origin “unlikely” these two markers are
immunohistochemically negative, can we do the same
interpretation in TNBC’s relying on these two markers?
In this cross sectional study, we collected 66 archived paraffin
blocks previously diagnosed as primary basal-like TNBC
in Mehr Hospital (Tehran, Iran). Immunohistochemical
studies for ER, PR, HER2, cytokeratin 5/6 and vimentin
were previously performed on all cases between the years
2005 and 2012. All of these cases were sectioned in a fresh
state and fixed overnight in 10% neutral buffered formalin
not longer than 24 hours. All tissue blocks were used by the
Tissue microarray: Tissue microarray (TMA) (Tissue
arrayer Minicore, Alphelys, Plaisir, France) was made of 66
paraffin blocks of TNBC’s. Three 1.4 mm tissue cores were
obtained from tissue blocks on each case for increasing
confidence coefficient. Each TMA contained 66 tissue
cores, each 1.4 mm in diameter. These were arranged as 7
rows and 10 columns with 2 positive and negative controls
Immunohistochemical Analysis for Mammaglobin and
GCDFP-15: Immunohistochemical studies for GCDFP
and mammaglobin were performed on TMAs. Fourmicrometer-
thick formalin-fixed, paraffin-embedded
sections were immunostained with mammaglobin and
GCDFP-15 antibodies (FLEX Monoclonal Mouse Anti-
Human Mammaglobin Clone 3041A5, FLEX Monoclonal
Mouse Anti-Human Gross Cystic Disease Fluid Protein-15,
Clone 23A3, Ready-to-Use, DAKO, Glostrup, Denmark).
The procedure was composed of pretreatment tissues using
EnVision FLEX, Target Retrieval Solution, PH 8.0 followed
by incubation with mammaglobin and GCDFP-15 mouse
antibodies. Counterstaining in hematoxylin was used by
EnVision FLEX Hematoxylin.
Positive and negative controls were run simultaneously
using the same protocol as the patient specimens. For
meaningful semiquantitative analysis, focal and/or weak
staining was considered negative staining, and only patchy
or diffuse staining with moderate or strong intensity was
Staining with the mammaglobin antibody was
characteristically seen in the cytoplasm. The staining
intensity in our series ranged from weak to moderate or
strong. The number of cells stained with the antibody was
further categorized as focal (<10%), patchy (10%-50%), and
diffuse (>50%) (Table I
Mammaglobin marker was positive only in 6 cases (9.1%)
and the rest 60 cases (90.9%) were negative (Figure 1A,B).
Immunohistochemically, GCDFP-15 marker was positive
in 12 cases (18.2%) and the other 54 (81.8%) remained
negative (Figure 2A,B).
Click Here to Zoom
|Figure 1: Basal-like triple negative breast carcinoma (A, H&E, ×400) showing strong staining with mammaglobin (B, ×400).
Click Here to Zoom
|Figure 2: Basal-like triple negative breast carcinoma (A, H&E, ×400) showing strong staining with GCDFP-15 (B, ×400).
Breast cancer is the most common cancer in women in many
countries of the world and is also the second cause of death
due to cancers after lung cancer20
. TNBCs are subgroups
of breast cancers identified by negativity for ER, PR and
Her2/neu receptors. These cancers have a short survival
and high metastatic rate7
. Although mammaglobin and
GCDFP-15 have been discovered almost a decade ago, few
studies have discussed their benefit for diagnosis of TNBCs.
Interestingly, mammaglobin is positive in few melanomas,
and it is important to recognize this pitfall, for, it is unlikely
to cause a problem in differential diagnosis from a breast
carcinoma because a panel of immunohistochemical stains
is always used in this matter. Mammaglobin does not
seem to be a useful stain to distinguish breast from sweat
gland carcinomas. Positive mammaglobin staining seen in
salivary gland tumors was also expected; however, intense
staining was not seen in any of the positive salivary gland
In Gloria H. Lewis’ study, the sensitivity of mammaglobin
and GCDFP-15 IHC markers for detecting basal-like
TNBCs has been 24% and 5%, respectively. In our study,
9.1% and 18.2% of basal-like TNBC cases showed posistive
immunohistochemical reaction for mammaglobin
and GCDFP-15. Unlike Lewis et al’s study, GCDFP-15
immunostaining was more frequent than mammaglobin
immunostaining in our study.
Therefore, the false negative results of these markers for
detecting TNBC’s rated as 90.9% and 81.8% respectively.
Hence, during pathological workup of a metastatic tumor
with unknown origin by using these markers, the possibility
of a TNBC entity will presumably missed. Since diagnosing
a TNBC is a crucial factor in choosing a systemic treatment,
postponing it can lead to a significant tumor progression.
TNBCs are highly aggressive with more intention to
metastasize and shorter survival period. Our study revealed
that 2 of the best IHC markers, used to detect the breast
origin, have very low sensitivity. As these tumors metastasize
to distant organs there is no useful IHC marker to detect
the origin of the tumor. Therefore, it will be important for
future studies to focus on finding accurate IHC markers for
detection of this type of breast cancer and communicate
closely with clinician and radiologist to correlate with
We are grateful to pathology technicians of Tehran
University of Medical Sciences and Mehr hospital for their
assistance in processing and staining of the samples, Dr.
Elham Mogharrablou and specially Beheshte Roudiany
for her great assistance in the grammar of this article. The
study was funded by Tehran University of Medical sciences,
The authors declare that there is no conflict of interests.
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