Immunohistochemical localization of notch signaling molecules in ameloblastomas

Muraki E, Nakano K, Maeda H, Takayama M, Jinno M, Kubo K, Yoshida W, Hasegawa H, Kawakami, Kawakami T

Description

We examined Notch signaling molecules, Notch1 and Jagged1, in serial large cases of typical solid/multicystic ameloblastoma. In general, Notch positive staining products were frequently detected in the cytoplasms of the cells. In the same cells, Jagged positive staining were also frequently observed, while only occasionally positive in peripheral cells, especially in cuboidal cells. The results showed that these morphogenesis regulation factors are closely related to cytological differentiation in neoplastic cells of ameloblastoma. The Notch and Jagged positive-cell ratios were frequently positive, and the ratios were nearly the same between the varied histopathological, cytological patterns. However, the less-differentiated cells were fewer in number than that of well-differentiated cells.

Sujets

• Ameloblastoma
• Notch signaling pathway
• Cellular differentiation
• Immunohistochemistry

Informations

• Publié par: biomed
• Publié le: 01 janvier 2011
• Nombre de lectures: 4
• Langue: English
• Poids de l'ouvrage: 4 Mo

Extrait

3) Kawakami T_Umbruchvorlage 03.06.11 17:15 Seite 253
June 21, 2011 Eu Ro P E a n Jo u R n a l o F MEDI c a l RE SEa Rc H 253
Eur J Med Res (2011) 16: 253-257 © I. Holzapfel Publishers 2011
IMMu n o HISt o c HEMIc a l l o c a l Iz a t Io n o F n o t c H SIg n a l In g
Mo l Ec u l ES In a MEl o b l a St o Ma S
1 1,3 2 2 2 2 2 3E. Muraki , K. n akano , H. Maeda , M. t akayama , M. Jinno , K. Kubo , W. Yoshida , H. Hasegawa ,
1, 3t . Kawakami
1Hard t issue Pathology u nit, Matsumoto Dental u niversity Institute for o ral Science, Shiojiri, Japan
2Department of o ral Pathology, School of Dentistry, a ichi g akuin u niversity, n agoya, Japan
3Hard t issue Pathology u nit, Matsumoto Dental u niversity g raduate School of o ral Medicine, Shiojiri, Japan
Abstract ticed different features of n otch expression patterns
We examined n otch signaling molecules, n otch1 and in ameloblastomas, when comparing our data with that
Jagged1, in serial large cases of typical solid/multicys- of Kumamoto and o hki [5]. t herefore, in this paper,
tic ameloblastoma. In general, n otch positive staining we re-examined n otch signaling molecules in serial
products were frequently detected in the cytoplasms of large cases of solid/multicystic ameloblastoma.
the cells. In the same cells, Jagged positive staining
were also frequently observed, while only occasionally Ma t ERIa l S a n D MEt Ho DS
positive in peripheral cells, especially in cuboidal cells.
t he results showed that these morphogenesis regula- t he surgical materials of ameloblastoma examined in
tion factors are closely related to cytological differenti- this study were obtained from operations, and diag-
ation in neoplastic cells of ameloblastoma. t he n otch noses were carried out at the Department of o ral
and Jagged positive-cell ratios were frequently positive, Pathology, a ichi g akuin u niversity School of Den-
and the ratios were nearly the same between the varied tistry, n agoya, Japan. We chose a total of 50 cases of
histopathological, cytological patterns. However, the ameloblastoma from the recorded surgical files, and the
less-differentiated cells were fewer in number than 50 cases were histopathologically re-examined. a total
that of well-differentiated cells. of 40 cases of typical solid/multicystic amelob lastoma
[1] were selected. t he summarized clinical data of se-
Key words: a meloblastoma; n otch signaling; n otch; lected 40 cases of surgical material are shown in t able
Jagged; c ell differentiation; Immunohistochemistry 1. Male 24, female 16; maxilla 4, mandible 36, total 40;
and these mean age is 27.6 years old.
In t Ro Du c t Io n
Table 1. Summary of Surgical Materials Examined.
a meloblastoma is a benign but locally invasive poly-
morphic neoplasm consisting of proliferating odonto- Age Sex Location
genic epithelium, which usually has a follicular or plexi-
form pattern lying in a fibrous stromal tissue [1]. t here Mean 27.6 Male 24 Maxilla 4
are four basic histopathological variations: 1) solid/
Female 16 Mandible 36multicystic; 2) extraosseous/peripheral; 3) desmop lastic;
and 4) unicystic. Regarding the solid/multicystic type,
there are two basic histopathological patterns, the fol-
licular and plexiform. t he follicular pattern consists of Immediately after removal, the surgical materials were
islands of odontogenic epithelium within the fibrous fixed in 10 % neutral buffered formalin solution. t he
stromal tissue. t he peripheral cells of these islands are specimens were then dehydrated through a series of
columnar, and hyperchromatic, and they are lined up in ethanols, and embedded in paraffin. a fter sectioning,
a palisaded fashion [1]. the series specimens were examined by histopathologi-
n otch molecules act as implementation of differen- cal (HE) methods.
tiation, proliferation, and developmental processes. a fter histopathological examination, we examined
Furthermore, n otch activity causes association with a on the distribution of transcription factors of n otch1
wide range of developmental disorders of neoplastic and Jagged1 by immunohistochemical (IHc ) tech-
cytological differentiation [2-4]. Kumamoto and o hki niques. IHc examination was carried out using a
t M[5] have studied n otch signaling molecules using serial Da Ko EnVision +Kit (Dako c ytomation,
cases of ameloblastoma. We have also examined some g lostrup, Denmark) with the following 2 antibodies:
ameloblastomas [6-10] and other types of odonto- n otch1 rabbit polyclonal antibody (ab27526, a bcam
genic neoplasms, such as odontogenic myxoma, squa- plc, c ambridge; dilution: 1/1000; 4 °c , overnight) and
mous odontogenic tumor, calcifying cystic odonto- Jagged1 rabbit polyclonal antibody (ab7771, a bcam
genic tumor, and calcifying epithelial odontogenic tu- plc, c ambridge; dilution: 1/500; 4 °c , overnight). a s
mor [11-14]. In our serial examinations, we have no- pre-treatment of immunohitochemical staining using3) Kawakami T_Umbruchvorlage 03.06.11 17:15 Seite 254
254 Eu Ro PEa n Jo u Rn a l o F MEDIc a l RESEa Rc H June 21, 2011
the above mentioned Kit, autoclave pretreatment arranged in anastomosing strands/cords with incon-
(120 °c , 10min) for n otch and protease K (Room spicuous stellate reticulum cells.
temp, 2 min) for Jagged were applied. Diaminoben- t hrough immunohistochemical observation, there-
dizine (Da b) was applied for the visualization of IHc fore, we described each case using the following crite-
activity and counter staining was carried out by hema- ria. a summary of the immunohistochemical examina-
toxylin. We included IHc staining using phosphate tion ratings is shown in t able 2.
buffered saline in place of the primary antibody as a 1) Peripheral cuboidal cells of nests: In general,
negative control. n otch positive staining products were detected in the
For the objective rating of the immunohistochem- cytoplasms of the cells in frequently. In the same cells,
istry, the observation points were divided into the fol- Jagged-positive cells were occasionally positive, but
lowing: 1) peripheral cuboidal cells of nests, 2) periph- less so than n otch-positive cells.
eral columnar cells of nests, 3) central reticular cells of 2) Peripheral columnar cells of nests: n otch posi-
nests, 4) central squamous cells of nests, and 5) stro- tive cells were observed in the cytoplasm of most
mal fibroblasts. Each case of above-mentioned classi- columnar cells, but Jagged was frequently detected in
fication was measured. t hose one with positive reac- the cells.
tion regardless of dye strength were assumed to be 3) c entral reticular cells of nests: t he central retic-
positive. t he number of positive cells was totaled, and ular cells showed frequently positive to both of n otch
the ratio of the number of positive cells to the total of and Jagged.
the object cells of the strong enlarged image by a light 4) c entral squamous cells of nests: both n otch-
microscope was assumed to be the c S-index. t he and Jagged-positive cells were frequently detected in
mean value of -: negative, 0% cells; +: occasionally the squamous cells, as observed acanthomatous meta-
positive, less than 25% cells; ++: frequently positive, plastic cells, but negative and/or not detected in the
25%-70% cells; and +++: almost positive, grater than keratinizing cells.
70% cells, was calculated by each c S-index of the cas- 5) Stromal fibroblasts: n otch positive stromal fi-
es examined. broblasts were frequently scattered in the tissues, but
t he study was approved by the Ethics c ommittee Jagged was occasionally positive, as observed slightly
(Endorsement number #179, Date: May 29, 2009), less than n otch.
School of Dentistry, a ichi g akuin u niversity. 6) Special findings: Regarding the peripheral
cuboidal cells located in the strands and/or cords pat-
RESu l t S tern regions of the follicular nests, both n otch and
Jaggeed were mostly negative, but both positive in the
Histopathologically, the tumor cell nests of ameloblas- peripheral columnar cells of the typical follicular nests
toma were proliferated in the stromal fibrous tissues. (Fig. 1-D, g ). o n the other hand, both the n otch and
t he growth pattern of these cell nests showed some Jagged positive stainings were detected frequently in
follicular and some plexiform structures (Fig. 1-a , b, c ). the central area, but Jagged stainings were occasionally
t he former consisted of islands of odontogenic positive, slightly less so than n otch in the peripheral
epithelium. t ypically, the peripheral layered cells of layers (Fig. 1-E, H). o n occasion, squamous cells, such
these cell nests were columnar or cuboidal, hyperchro- as acanthomatous cells and keratinizing cells, were ob-
matic, and lined up in a palisade fashion. t he periph- served. t he acanthomatous squamous cells showed
eral layered cells resembled internal dental epithelium frequently positive (Fig. 1-F, I), but they were not de-
or preameloblasts. t he central cells were loosely termined in the keratinizing squamous cells.
arranged, showing stellate reticulum. t hese nests
sometimes became cystic formations. Furthermore, DISc u SSIo n
the central cells sometimes showed the cytological
change of squamous metaplasia. a meloblastoma is one of typical odontogenic benign
t he latter contains columnar and/or cuboidal cells neoplasm arising from epithelium of the odontogenic
apparatus and its remnants. Histopathologically, there