Expression of mucins in normal salivary glands and mucoepidermoid carcinoma of salivary glands. Matilda Llupi Rabije Qoku Supervisor: Gunnar Warfvinge, Dept. of Oral Pathology. Master Thesis (30 hp) Malmö University Dentistry Program Faculty of Odontology February, 2013 205 06 Malmö
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Expression of mucins in normal salivary glands and ... · and account for approximately 34, 20 and 30 % of the malignant tumours in parotid glands, submandibular glands and minor
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Expression of mucins in normal salivary
glands and mucoepidermoid carcinoma of
salivary glands.
Matilda Llupi
Rabije Qoku
Supervisor:
Gunnar Warfvinge, Dept. of Oral Pathology.
Master Thesis (30 hp) Malmö University
Dentistry Program Faculty of Odontology
February, 2013 205 06 Malmö
2
ABSTRACT
Mucoepidermoid carcinomas (MECs) are malignant epithelial mucin-producing tumours encountered
in both major and minor salivary glands. The aim of this study was to investigate the histological
characteristics of the expression of mucins (MUC1, MUC4, MUC5AC, MUC5B, MUC6) in MECs in
search for a possible correlation between qualitative mucin expression and tumour grade. Twelve low-
grade, five high-grade MECs and nine normal salivary glands adjacent to tumour tissue were
investigated for these mucins by immunohistochemistry. The samples were evaluated with respect to
staining pattern and positivity of specific cell types. Normal acinar cells mainly expressed the
cytoplasmic mucin MUC5B. MUC1 and MUC4 were expressed in normal ductal cells in approximately
half of the samples whereas MUC5AC expression was rare in normal salivary glands. MECs
expressed MUC1, MUC4, MUC5AC and MUC5B. The apical membrane of mucous cells lining the
cystic cavities showed the strongest staining for MUC1 and MUC4. The expression of MUC4 in
mucous cells decreased with increasing histological grade. Expression of salivary mucin MUC5B in
mucous cells in low-grade MECs was less intense compared to the expression of MUC5AC in the
same cells. In high-grade tumours, a higher expression of MUC5B compared to MUC5AC was noted.
In conclusion, MECs express different mucin quantity compared to normal salivary glands. MUC5AC
expression in salivary tumour tissue seems to be a metaplastic feature and MUC4 appears to be
related to tumour differentiation grade. The relationship between MUC5AC and MUC5B expression
could be a useful tool in the diagnosis and estimation of prognosis of MECs.
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TABLES
TABLE 1. Clinical outcome of MECs depending on histological grade (11).
TABLE 2. Antibodies used in the study.
TABLE 3. MUC5B and MUC5AC immunoreactivity in tumour cells in low- and high-grade MECs.
Low-grade Intermediate-grade High-grade
Local recurrence 6% 20% 80%
Metastasis exceptional rare frequent
5 year-survival 95% 80-90% 25-30%
Antigen/ Antibody
Origin Sequence Positive control
tissue Labeling Dilution
MUC1/ MUC1B
Polyclonal/rabbit/ C-terminal
CRRKNYGQLDIFPARD Breast cancer Membrane-bound
1/2000
MUC1/ 214D4
Monoclonal/mouse/ N-terminal
--------------------------------- Breast cancer Membrane-bound
1/500
MUC4/ MUC4-8
Polyclonal/rabbit/ C-terminal
WNNNPEDDFRMPC Adenocarcinoma from pancreas
Membrane-bound
1/2000
MUC5AC LUM5-1
Polyclonal/rabbit/ mucin domain
RNQDQQGPKFMC Normal gastric
mucosa Cytoplasmic 1/2000
MUC5B/ LUM5B-2
Polyclonal/rabbit/ mucin domain
RNRWQVGKFMC Normal salivary
gland Cytoplasmic 1/8000
MUC6/ LUM6-3
Polyclonal/rabbit/ mucin domain
RPLHSYEQQLELPC Normal gastric
mucosa Cytoplasmic 1/4000
MUC5B MUC5AC
Low-grade High-grade Low-grade High-grade
Negative 1 0 1 3
<50% of tumour
cells 7 3 1 2
≥50% of tumour
cells 4 2 10 0
N = 12 N = 5 N = 12 N = 5
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FIGURES
FIGURE 1. Proposed histogenesis of MEC (modified from 3).
FIGURE 2. A. Appearance of low-grade MEC. Note the mucous cells (arrow) lining the cystic cavities.
B. Intermediate-grade MEC. C. A solid high-grade MEC mainly containing intermediate cells. D. A
mixture of mucous cells (1), intermediate cells (2) and epidermoid cells (3) in low-grade MEC. (HTX,
original magnification A,B,C x 20; D x 40)
1
2
3
A B
C D
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FIGURE 3. Expression of mucins in acinar and ductal cells of normal salivary glands adjacent to
tumour tissue (n = 9 patients).
FIGURE 4. Representative examples of differences in staining pattern for MUC5B and MUC5AC.
A. A section from a low-grade MEC stained for MUC5B. Note a weak positively stained mucous cell
(arrow). B. A serial section from the same low-grade MEC (A) stained for MUC5AC. The mucous cells
show strong positivity for MUC5AC (arrow). C. A high-grade MEC positively stained for MUC5B. D.
The same high-grade MEC (C) negatively stained for MUC5AC. (original magnification x 40).
0
2
0
9
1
4
5
1
0
9
0
2
4
6
8
10
MUC1 MUC4 MUC5AC MUC5B MUC6
Po
siti
ve c
ases
(n
um
ber
)
Acinar cells
Ductal cells
A B
C D
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FIGURE 5. Representative examples of staining pattern for MUC4 in low-grade MEC (A) and high-
grade MEC (B). MUC4 stains the apical membrane of mucous cells in low-grade MEC (arrow in A).
The staining in high-grade MEC is also specific but weak (arrow in B). (original magnification x 40).
FIGURE 6. Staining pattern for MUC1, MUC4 and MUC6 in mucous and epidermoid cells of MECs