2018, Volume 34, Number 3, Page(s) 220-224
Fascin Expression in Ameloblastoma, Odontogenic Keratocyst and Dentigerous Cyst
Sedigheh RAHROTABAN1, Nariman NIKPARTO2, Parham YOUSEFIZADEH3, Mohammad Javad KHARAZIFARD4, Samira DERAKHSHAN1
1Department of Oral and Maxillofacial Pathology, Tehran University of Medical Sciences, School of Dentistry, TEHRAN, IRAN
2Oral and Maxillofacial Surgery Resident, Tehran University of Medical Sciences, School of Dentistry, TEHRAN, IRAN
sup>3Dentist, Private Practice, TEHRAN, IRAN
4Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, TEHRAN, IRAN
Keywords: Ameloblastoma, Dentigerous cyst, Fascin, Odontogenic keratocyst
The purpose of this study was to assess and compare fascin expression in 4 lesions which differ in aggressiveness: odontogenic
keratocyst, dentigerous cyst and two types of ameloblastoma (solid and unicystic), and to find out whether fascin expression is associated with
aggressiveness of these lesions or not.
Material and Method: Nine solid ameloblastomas , 12 unicystic ameloblastomas, 13 odontogenic keratocyst and 12 dentigerous cyst were
assessed in this study. The slides were examined at x400 magnification. Finally the lesions were divided into two groups based on microscopic
examination, “low expression” and “high expression”.
Results: There were no significant differences between the lesions, except that fascin expression was slightly higher in unicystic ameloblastomas
in comparison to other groups in intensity and count of the immunostaining cells.
Conclusion: The results of this study suggest that local aggressiveness does not result in fascin expression. We suggest more studies with more
samples, assessing expression of different proteins be done in the future.
Fascin is a known 55-kDa globular actin-bundling protein
which is mainly expressed in cell membrane protrusions
like microspikes called filopodia. This protein has three
isoforms in mammals: Fascin-1 (referred to from here
on as Fascin) which is highly expressed in neural and
mesenchymal tissues during embryogenesis and in brain,
endothelium and testis in adults, fascin-2 in retina and
fascin-3 in testes 1-3
. Fascin is mainly expressed in
neurons, dendritic cells and pericytes which have striking
large filopodia and which are highly mobile 4-6
. Also this
protein is expressed in endothelial cells and fibroblasts 7,8
. Fascin expression has been found to be low or absent
in the majority of normal adult epithelial tissue varying
origin. However, in stratified squamous cells of skin, fascin
is expressed at a very low intensity and only in the basal
. Frequent and increased fascin expression in
healthy, although tumor-adjacent, epithelial tissue has
previously been reported 10
Epithelial cell junctions are mostly dependent on cellcell
or cell-matrix interactions which help to stabilize the epithelial cells and prevent their migration. In epithelial
malignancies, invasion to basal lamina and metastasis
occurs due to elimination of these junctions. Fascin is
considered in the migration and invasion of carcinoma
cells since this actin-cross-linking protein is known to
be responsible for controlling the guiding system of cell
Many studies have reported the upregulation of fascin
expression in various tumors. Moreover it has been
reported that high expression of fascin is associated with
high motility of cells, poor differentiation, advanced grade
and stage and metastasis 11,12. Fascin helps to stabilize
the actin bundles in protrusions and it therefore seems to
be used by cancerous cells to assemble stable and durable
invasive protrusions (called invadopodia) that help
invasion into the matrix 13,14.
Ameloblastoma is slow growing and locally aggressive
odontogenic epithelial tumor with a low tendency to
metastasize and a high recurrence rate 15.
Odontogenic keratocyst (OKC), formerly known as
keratocystic odontogenic tumor (KCOT), is a common odontogenic cyst that originates from the dental lamina or
from its residue; occurring at any age, and is more common
in men than women with a 2:1 ratio 16). Because of the
high proliferation rate, aggressiveness and recurrence rate
of epithelial cells of this cyst, in 2005 WHO reclassified
odontogenic keratocyst as benign intra-osseous neoplasia,
called KCOT 17. However, this odontogenic lesion is now
listed as odontogenic keratocyst in the 2017 classification
of developmental odontogenic cysts 16.
Dentigerous cyst (DC) is the second most common
odontogenic cyst (following radicular cyst) which is
typically seen as a radiolucency surrounding the crown of an
impacted tooth. DC is not as aggressive as ameloblastoma
or OKC 18.
In this article, we assessed and compared fascin expression
in 4 lesions which differ in aggressiveness: OKC, two types
of ameloblastoma (solid -which is believed to be more
aggressive- and Unicystic) and DC to find out whether the
fascin expression is associated with aggressiveness of these
lesions or not.
Due to the retrospective nature of this study, it was granted
an exemption in writing by the institutional review board
of Tehran University of Medical Sciences (TUMS). For this
descriptive analytic study, 82 biopsy-proven samples were
retrieved from the archives of Departments of Oral and
Maxillofacial Pathology of Tehran and Qazvin University
of Medical Sciences. A total of 36 cases were excluded
because of quantitatively inadequate available tissue in
paraffin blocks, and recurrent or inflammatory lesions.
Thus, 46 paraffin blocks including 9 solid ameloblastomas
(SA), 12 unicystic ameloblastomas (UA), 13 OKCs and 12
DCs were assessed in this study.
After obtaining sections with a thickness of 4μm,
immunohistochemical staining was carried out with the
monoclonal anti-fascin antibody (Clone 55K-2 Dako,
Glostrup, Denmark) diluted 1:50. For each case, a section
with the primary antibody omitted was used as negative
control and the cytoplasm of endothelial cells was used as
internal positive control.
In microscopic assessment, the slides were examined at x400
magnification (Figure 1A-F) by an oral and maxillofacial
pathologist who was blinded to the clinical characteristics
of the samples. Immunoexpression of fascin in tumor cells
was evaluated in terms of extent and intensity. The extent
of immunostaining was scored according to Terence K. Lee
19: 1-negative: less than 10% of cells, 2-weak: 11-50% of
cells, 3-moderate: 51-80% of cells and 4-strong: more that
81% of cells. Moreover, the staining intensity of the samples was categorized according to the cytoplasmic staining
of cells into: 1-faint, 2-moderate and 3-severe. Finally,
to determine the expression level of fascin, a Combined
Immunoreactivity Score (CIS) was calculated by adding
the score of extent to the score of intensity for each case.
The samples were further classified into 4 groups: Negative
expression: staining in less than 10% of cells regardless
their staining intensity, weak expression: CIS equal to 3,
moderate expression: CIS equal to 4 or 5, strong expression:
CIS equal to 6 or 7.
Click Here to Zoom
|Figure 1: A) High expression of fascin in basal and suprabasal layers in odontogenic keratocyst. Note immunoactivity of stomal fibroblasts
and endothelial cells as positive internal control (IHC; x100). B) Low expression of fascin only in basal layer in odontogenic keratocyst
(IHC; x100). C) High expression of fascin in basal and suprabasal stratum reticulum layers in unicyctic ameloblastoma (IHC; x400).
D) Low expression of fascin in unicyctic ameloblastoma. Note immunoactivity of stromal fibroblasts and endothelial cells even in low
epithelial lining reaction (IHC; x100). E) High expression of fascin in dentigerous cyst (IHC; x100). F) Low expression of fascin in
dentigerous cyst (IHC; x100).
In statistical analyses, the first 2 groups were merged as the
“low expression” group, and the groups 3 and 4 merged as
the “high expression” group. Research data were analyzed
using the Statistical Package for Social Sciences (SPSS),
version 18. The Kruskal-Wallis and two by two Dunn
Tests were used. P values less than 0.05 were considered
The scores of stained-cell counts, staining intensities and
CIS are shown in Table I
. Ten samples from 12 unicystic
ameloblastoma (83.3%) showed a score of 3 in intensity
and also 10 samples showed a score of 4 in the count of
immunostaining cells (Table I
). As mentioned formerly,
these four groups were merged into low expression (group
1&2) and high expression (group 3&4). No significant
difference was found between these groups (P>0.05). All
the samples of the four groups (DC, OKC, SA and UA) were
classified in the high expression group except one sample of
OKC (7.7%) that was classified in the low expression group
). In the two by two comparison test, although a
significant P value was seen in UA comparison with other
lesions, there was no significant difference between the
groups when other criteria were considered in the statistical
Click Here to Zoom
|Table I: Scores of stained-cell counts, staining intensities and group classifications based on immunohistochemical assessments
Four different tumors based on aggressiveness of their
clinical behavior were studied in this study. The aggressive
behavior of OKC was close to ameloblastoma as in many
studies which investigated the expression of markers such
as p53, p63, Ki67, PCNA, AgNors and Ipo 20-23
concluded that the proteins expressed on epithelial cells of
OKC is more alike to that of ameloblastoma than DC.
In order to assess aggressiveness of lesions, some studies
have examined the extra cellular matrix and connective
tissue proteins such as fibronectin, laminin, tenascin,
RANK, RANKL, and OPG in these lesions. Many of them
found significant differences in the expression levels of
these proteins among DC, ameloblastoma and OKC 24-27.
All mentioned markers are exclusively expressed in
connective tissue cells except laminin which is expressed in
epithelial tissue. In this study we assessed fascin, as another
epithelial cell marker. Fascin is also known as the protein
of motility 11,12. No wonder that expression of fascin
can be associated with aggressive and invasive behaviors
28,29 as seen in many epithelial cancers 13. Fascin is
believed to be a biomarker that indicates aggressiveness and
migration of tumors. However, our results contradict this
hypothesis as we observed no significant upregulation of
fascin in aggressive lesions. In addition, one meta-analysis
study demonstrated that fascin may have potential as a
novel biomarker for early identification of aggressive and
metastatic tumors 12. In our study, the only statistically
significant difference among the four lesions was the
increased fascin expression in unicystic ameloblastoma in
comparison to the other three. There is a possibility that
fascin might be more expressed in carcinomas than cyst
walls or odontogenic tumors. Also since the lesions of this
study were mostly intra-bony, there were no epithelial
barriers to their progression. Moreover these lesions do
not metastasize. Considering these, we suggest that fascin
might be more involved in cell invasion and migration (as in
carcinomas) than local aggressiveness. Lee et al. suggested that fascin over-expression might enhance oral squamous
cell carcinoma aggressiveness, possibly by interacting
with E-cadherin expression 19. There is no E-cadherin
interaction in progression of odontogenic lesion versus
progression of carcinomas. Therefore, according to Lee
theory 19, our suggestion based on more involvement of
fascin in invasion and migration than local aggressiveness
in benign odontogenic lesion is justifiable. Furthermore,
Li et al. showed that fascin is an integral component of
invadopodia -the finger-like protrusions using by cancer
cells which help them invade into and degrade extracellular
matrix- and it is important for the stability of actin in
invadopodia 28. Epithelial cells of benign odontogenic
lesions like DC, OKC or ameloblastoma that we assessed
in this study do not use invadopodia for progression, and
this may be a reason for less involvement of fascin in local
It is accepted that immunohistochemistry is a complex
metric due to the use of different scoring systems to
assess the extent of staining in different specimens but
the scoring of fascin is a continuous measurement and in
most publications researchers categorized specimens into
high/positive or low/negative fascin based on different
semi-quantitatively assessed cut-off points 12. We also assessed the fascin expression in two groups of low and
high expression as in most previous publications. However,
this study has several limitations. First, we did not assess
whether these lesions had perforated the cortical bone and
invaded local soft tissue. Second, we just examined fascin
expression and not other proteins or markers. We suggest
more studies with more samples, and assessing expression
of different proteins in the future.
CONFLICT of INTEREST
The authors declare no conflict of interest.
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