Download Predominant Expression of Human Zic in Cerebellar Granule Cell

CANCERRESEARCH
56. 377-383.January 5. 19961
Predominant Expression of Human Zic in Cerebellar Granule Cell Lineage and
Medulloblastoma'
Naoki Yokota,2 Jun Aruga,
and Katsuhiko Mikoshiba
Setsuo Takai,
Kiyomi
Yamada,
Minoru
Hamazaki,
Toshio
Iwase,
Haruhiko
Sugimura,
Molecular Neurohio!ogt' Laboratory, Tsukuba Life Science (‘enter,
The Institute of Physical and Chemical Research (RIKEN.1, 3-1-I Koyadai, Tsukuba. Ibaraki 305 [N. Y.. J. A..
K. MI; Department of Microbiology and immunology (N. Y.J and 1st Department of Pathology (T. I., H. S.], Hamamatsu University School of Medicine, 3600 Handa-cho,
Hamamatsu, Shizuoka 431—31:Department of Pathology, Shizuoka Children ‘s
Hospital. 860 Urushivama, Shizuoka 420 (Y. HI; Department of Genetics, Research institute,
international Medical Center of Japan. 1-21-I Tovama-cho, Shinjuku-ku, Tokyo 162 (5. T., K. Y.J; and Department of Molecular Neurobiology, institute of Medical Science, The
University of Tokyo. 4-6- 1 Shirokanedai, Minato-ku, Tokyo 108 (K. MI, Japan
expressed in the granule cell lineage throughout the cerebellar devel
ABSTRACT
opment
Zic is a novel zinc finger protein which displays a highly restricted
expression
pattern
in the adult
and developing
mouse cerebellum
and is
highlyhomologous
to the recentlyclonedDrosophilapair-rule geneOpa.
To clarify the mechanismfor the developmentof the human cerebellum
and its involvementin humannervoussystemdiseases,
we haveisolated
human Zic eDNA and examinedits expressionby using monoclonal
antibodyagainstrecombinantZic protein.The nucleotidesequenceof
humanZic cDNA is 85% homologousto that of mouseZic eDNA. Its
putative amino acid sequenceis highly conserved (>99%) except for
substitution of only two amino acid residues.In situ chromosomehybrid.
in the
mouse
(9).
Its zinc
finger
domain
shows
a high
homology to the Drosophila pair-rule gene Opa (10), the Caeno
rhabditis elegance sex determination gene tra-1 (1 1), the Drosophila
segment
polarity
gene
ci'@ (12),
and
the
human
oncogene
Gil
(13).
Such similarities and expression patterns suggest that Zic plays a role
in the developmentalprocess.Furthermore,Zic is potentiallyinvolved
in the oncogenesisof undifferentiatednervoussystemtumors,includ
ing medulloblastoma, since several oncogenes, such as Writ-I and
GLI, exhibit expression in a spacially and temporally restricted pattern
and play a role in the early organization of the structure (14, 15).
izationlocalized
thehumanZicgeneto chromosome
band3q24.Human In the present study, to clarify the histogenesis of the human
Zic protein was immunohistochemically
detectedin the nuclei of the cerebellar granule cell lineage and its involvement in human nervous
cerebellar granule cell lineage from the progenitor cells of the external
system diseases,especially in medulloblastoma, we cloned human Zic
germinallayer to the postmigratedcellsof the internal granular layer.
cDNA
and determined the chromosomal localization of the Zic gene
Furthermore, Zic protein wasdetectedin medulloblastoma(26/29cases),
by FISH. We also investigated its expression immunohistochemically
whereas no other tumors examined (over 70 cases including
primitive
neuroectodermal
tumors)
expressed
this protein.
These findings
in both normally
suggest
that Zic is a potentialbiomarker for medulloblastomaas well as the
humancerebellargranulecell lineage.
developing
human
cerebella
and various
pathologi
cal specimens of human tumors. In addition, the diagnostic potential
of immunohistochemical Zic protein in differentiating medulloblas
toma from other tumors is discussed.
INTRODUCTION
MATERIALS
The mammalian central nervous system contains an enormous
number of neurons categorized into many subtypes. One of the major
subjects of study is to understand the mechanism underlying the
generation of these neuronal subtypes. Our interest has been in the
processes of differentiation
and maturation
of cerebellar granule cells.
Becauseof their unique maturation processesand mitotic ability in the
postnatal period, we consider cerebellar granule cells an ideal system
in which to analyze the neurobiological development.
Another intriguing aspect of cerebellar granule cells is their in
volvement in a certain type of brain tumor, medulloblastoma. Medul
loblastoma, originally named by Bailey and Cushing (1), is a malig
nant brain tumor, and accounts for approximately up to 25% of
intracranial tumors in childhood (2). The origin of medulloblastoma
has been ascribed to three main sources: fetal granular layers identical
to the EGL3 (3), a “germinalbud―situated at the hind end of the
posterior medullary velum (4), and the IGL (5—7).
Medulloblastoma is
categorized to cerebellar PNETs (8); however, the origin of this tumor
is still controversial, and this category must be distinguished from
other undifferentiated neuroepithelial tumors.
We recently isolated a novel zinc finger protein, Zic, which is
Received 8/2 1/95; accepted I 1/9195.
The costs of publication of this article were defrayed in part by the payment of page
charges. This article must therefore be hereby marked advertisement in accordance with
18 U.S.C. Section 1734 solely to indicate this fact.
I This
study
was
supported
in
part
by
grants
from
the
Institute
of
Physical
and
Chemical Research (RIKEN), Science and Technology Agency, Japanese Ministry of
Education, Science, and Culture, and Japan Intractable Disease Foundation.
2 To
I The
whom
requests
abbreviations
for
used
reprints
are:
should
EGL,
be
extemal
layer;
IGL,
internal
eDNA and Genomic DNA Cloning. A cDNA libraryof humancerebellum
in AgtI 1(ClontechLaboratories),whosemRNA sourcewasnormalcerebellar
tissuefrom a 15-year-oldCaucasianfemale,wasscreenedat high stringency
using 32P-labeledmouseZic cDNA containingalmostthe entirecoding region
of Zic. Plaque lifts and subsequent purifications were performed essentially as
describedby Sambrooket al. (16). Sequencingreactionsand nucleotide
sequenceanalyseswereperformedaspreviouslydescribedby Arugaet a!. (9).
The complete
nucleotide
sequence
of three overlapping
clones
was determined
on both strands.
Using a human Zic cDNA
clone as a probe, a genomic
DNA
library
of
Japaneseadult humansin EMBL3 provided by the the JapaneseCancer
ResearchResourceBank(L1018)wasscreenedat high stringencyasdescribed
above. The clones obtained were analyzed by restriction mapping and partial
sequencing.
Genomic Mapping.
R-banded chromosomes were prepared using standard
methods (18) with some modifications
over 10 kb, which contains
(19). The Zic genomic clone (pghzic8,
almost the entire region of the human Zic gene) was
used as a probe for FISH. The probe was labeled with biotin-16-dUTP
(Boehringer) by nick translation. Hybridization
was performed as reported (19,
20).A 10-foldexcessof totalhumanDNA wasaddedto thehybridization
solution to suppressnonspecifichybridization. Signalswere detectedwith
fluoresceinavidin DCS (Vector Laboratories).The slideswere stainedwith
propidiumiodide(0.5 .&g/ml;Sigma)andexaminedundera Nikon Optiphot
2-EFD2 microscope (B2A filter). Kodak Ektachrome film (ASA 100) was used
for microphotography of the chromosomes.
Generationof mAb. The mouseglutathione-S-transferase—Zic
fusionpro
tein was isolated as described previously (9). BALB/c mice (6—8weeks old)
addressed.
germinal
AND METHODS
granular
layer; FISH. fluorescencein situ hybridization;mAb, monoclonalantibody;GW, gesta
tional week:PNET,primitive neuroectodermaltumor.
wereimmunizedwith thefusionprotein.Themicereceivedaninitial injection
of 100 i.@gprotein emulsified I:1 with Freund's complete adjuvant. Three
weeks later, the mice were given a boost of 100 @tg
protein in Freund's
incomplete adjuvant. Two weeks after the boost, the serum samples were
377
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HUMANZic cDNACLONINGAND ITS EXPRESSIONIN VARIOU5TUMORS
testedusinganELISA. Mice yieldinggoodserumresponses
wereboostedwith
100 @xg
proteinwith PBS3 daysbeforethe fusion.Spleencells werefusedto
5p2/O-Ag14myelomacells providedby Instituteof Physicaland Chemical
Research
(RIKEN)
Cell
Bank
(RCB2O9)
in the presence
of
PEG4000
(GLBCO), using standard protocols (21). Hybridoma supernatants were
screened 1—2
weeks later with ELISA, immunohistochemistry, and immuno
blot stripscontainingtheglutathione-S-transferase—Zic
fusionprotein.Positive
clones were cloned by limiting dilution. Ascites was produced as described
human Zic as a homologue of mouse Zic cDNA and determined the
entire nucleotide sequence of 3. 1 kb. The human Zic nucleotide
sequence and deduced amino acid sequence are deposited in DNA
Data Bank of Japan, European Molecular Biology Laboratory, and
Genebank with the accession no. D76435. The human Zic nucleotide
sequence and its putative open reading frame display 85 and 95%
homology,
respectively,
with the mouse
sequence.
The open reading
frame is 1344 bp (477 amino acids) long preceded by 786 nucleotides
(21). The specificity of mAbs was checked by Western blot analyses.
Immunohistochemistry. Formaldehyde-fixedparaffin-embeddedsections of an untranslated region and has a calculated molecular mass of 48.4
(3—6 @m)
were preparedusing the usual procedures.The sectionswere kDa. Compared with the mouse Zic sequence,the human Zic-deduced
dewaxed in xylene and hydrated in a series of ethanols. Since tissue antigens
amino acid sequenceis highly conserved (>99%), with substitutions
were markedlydecreasedin the tumor specimens,they were autoclavedin at only two amino acid residues,Ser―@
to proline, and Met422to
0.01% citrate buffer (pH 6.5) at 110°C
for 10 mm as a meansof antigen
leucine. In the zinc finger region sparing 227—384,the human Zic
retrieval.The sectionswereincubatedwith 0.5%skim milk, 0.05%Tween20,
50 mMTris-HCI (pH 7.4),and 150mMNaC1for 60 mm at roomtemperature, amino acid sequence is identical to that of mouse Zic.
Northern blot analysis indicates that the size of mRNA is 3.1 kb,
followedby incubationwith anti-Zic mAb overnightat 4°C.
Immunoreactivity
which corresponds to the size of the cDNA. Whereas mouse Zic
wasdetectedby an immunoperoxidasereactionusingthe materialsand pro
ceduresprovidedin theVectastainkit (VectorLaboratories).3,3'-diaminoben mRNA consists of three different classes of cerebellar mRNA (2.9,
zidinetetrahydrochloridewasusedasthe substrateof theperoxidasereaction. 3.2,and5.0kb), only oneclassof mRNA(3.1kb) wasdetectedin
Zic Expressionin Developingand DevelopedHuman Cerebella.We humans (data not shown). This suggeststhat our cDNA clones cover
examined
immunohistochemically
Zicexpression
in pathological
specimens
of an almost full length of the mRNA.
the normally developingcerebellaof eight prematureinfantsat GWs 19—37 The Human Zic Gene Was Mapped to 3q24. Southern blot
and in the developedcerebellumof a 6-year-oldchild.
analysis of human genomic DNA extracted from peripheral blood
Zic Expressionin VariousHuman Tumors. Zic expression
wasfurther
mononuclear cells revealed that Zic is a single gene. A Zic genomic
examined in various human tumor specimens obtained from 29 cases of
clone
(phzic8; see “Materials
and Methods―)which contains almost
medulloblastoma,I 1 casesof glioblastoma.10 casesof retinoblastoma,10
the entire human Zic gene (data not shown) was used as a probe for
cases of ependymoma, 3 cases of Ewing sarcoma, 2 cases of choroid plexus
papilloma,I caseof extracranialPNET,and 1caseeachof hemangioblastoma, FISH analysis. Forty-five of 50 (pro)metaphases observed showed
ganglioneuroma,
ganglioblastoma,
malignant Schwannoma, alveolar soft part
symmetrical double spots on at least one homologue of chromosome
sarcoma,york sactumor,liposarcoma,immatureteratoma,giantcell sarcoma, 3. The signal region was localized to 3q24 (Fig. 1). No other chro
andpineoblastoma.
mosomes
exhibited
double
spot signals.
Thus, it was concluded
that
Zic expression was also looked for in a series of common carcinomas of
epithelialorigin, such as gastric(six cases), colorectal(five cases), liver (four
cases),breast(five cases),andlungcancer(six cases).Two casesof smallcell
carcinomaandfour casesof carcinoidtumorswerealsoexamined.
RESULTS
The Human Zic SequenceIs Highly Conserved.We obtained
three
overlapping
cDNA
clones
encompassing
complete
lengths
of
the human Zic gene is located at human chromosomal region 3q24.
Several linkage analyses of degenerative cerebellar atrophies have
been reported (23—28).However, none of the loci of the genes for
degenerative nervous diseases, clarified by these linkage analyses,
were identical to that of Zic.
Zic Was Expressed in Human Premature Infantile and Ma
hired Cerebella. A mAb, ZC26, could recognize Zic protein well
using Western blot analyses and showed a good immunohistochemi
1@,
4@ ...
4
Fig. I . The human Zic localization to human
chromosome 3q24 by FISH with R-banded chro
mosomes. Arrows, spots of the signals after FISH
using biotinated Zic genome DNA.
‘@@1
.
4
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Fig. 2. Immunohistochemical studies of Zic protein in human developing and developed cerebella. The cerebellum of a 21-OW premature infant: a, H&E stain. b, anti-Zic stain.
The cerebellum of a 37-OW premature infant: c, H&E stain. d, anti-Zic stain. The cerebellum of a 6-year-old child: e, H&E stain. f, anti-Zic stain. Note that the nuclei of the cells
in the EGL, inward-migrating granule cells, and postmigration cells in the IGL are strongly stained by anti-Zic immunohistochemical stain in 21 OW. The numbers of these
inward-migrating cells decreased in 37 OW; however, anti-Zic-positive cells in the EGL were still persistent. In the developed cerebellum, the migration of granule cells is over, and
the EGL has disappeared. Purkinje cells showed no reactivity against anti-Zic antibody in any stages.E, external germinal layer. P. Purkinje cell; G, internal granular layer; M. molecular
layer. X80. Bar, 100 @m.
379
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HUMAN Zic cDNA CLONING
AND ITS EXPRESSION
Table I hnmunohistochemistr, of anti-Zic mAb in various human tumors
in Table
Brain tumors
Medulloblastoma
Extracranial tumors
Ewing sarcoma
PNET
26/29
0/3
0/I
Other intracranialtumors
0/10
0/I
0/2
0/10
0/I I
0/3
0/I
Retinoblastoma
Pineoblastoma
Choroid plexus papilloma
Ependymoma(ependymoblastoma)
Glioma (glioblastoma)
Ganglioneuroma (ganglioblastoma)
Hemangioblastoma
Other cancers
Gastriccancer
a Number
of Zic-positive
cases/number
0/4
0/I
0/I
0/I
0/I
0/1
0/1
0/I
of examined
is a phylogenically
cases.
cal reactivity. It was also revealed that the mAb cross-reacted with
other members of the Zic family because of high homology of their
carboxy terminus.4 However, their expression patterns were almost
the same as that of Zic protein, and the amount of expression was
quite low according to Northern blot analysesand in situ hybridization
studies.
Furthermore,
the immunohistochemical
localization
of Zic
protein in rat central nervous system using this mAb correlated well
with the results of previous in situ hybridization studies in mice and
immunohistochemical
studies using the polyclonal antibody5 (9).
Thus, we used this mAb in the subsequent immunohistochemical
analyses. A four hundred-fold dilution of ZC26 appeared to result in
distinctive nuclear staining without background.
Zic protein was strongly detected in the nuclei of EGL component
cells and in those of inward-migrating granule cells and postmigrated
granule cells in the IGL, both in the developing and developed
cerebella (Fig. 2). These results indicate that human Zic protein is
predominantly expressed in the nuclei of the cerebellar granule cell
lineage throughout development.
Expression of Zic Protein in Human Tumors. Table 1 summa
rizes the profile of the results of immunostaining in various human
tumors, and we reviewed all cases of medulloblastoma, Ewing sar
coma, and PNET concerning clinical information such as age, sex, and
location in Table 2. Immunohistochemical positiveness of Zic protein
was highly
restricted
to cases of medulloblastoma.
A fraction
in the cells with round nuclei
and fine
chromatin structure, whereas the component of pyknotic tumor cells
and the cells with neuronal or glial differentiation were weak or
negative (Fig. 3). Three casesof medulloblastoma (cases7, 9, and 11
4 .1. Aruga,
T.
Nagai,
T.
Tokuyama,
Y.
Hayashizaki,
Y.
Okazaki,
V.
M.
immunoreactivity
was
Chapman,
gene that may participate
in an indis
Medulloblastoma
caseAgeSexLocation1
yr
2
3
4
10
5
yr
M
M
M
Vermis
Vermis
Vermis
10 yr
F
Vermis
6
7
8
9
10
11
1
11
9
4
9
8
yr 8mo
yr 4mo
mo
yrllmo
mo
yrlOmo
12
6
yr
M
M
M
M
M
F
M
M
F
F
F
M
F
M
M
F
M
M
F
M
F
F
F
FVermis
Vermis
Vermis
Vermis
Vermis
Hemisphere
Vermis
Vermis
Vermis
Posterior fossa
Vermis
Hemisphere
Vermis
Vermis
Vermis
Vermis
Vermis
Vermis
Vermis
Vermis
Hemisphere
Hemisphere
Hemisphere
Vermis
yr
yr
yrF
F
MThigh
Rib
Pelvis
yr
M
Submandibular region
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
HemisphereEwing
298
sarcoma
I
2
316
and
odd-paired, manuscript submitted for publication.
N. Yokota, J. Aruga. and K. Mikoshiba. Immunohistochemical localization of Zic
protein in the central nervous system of developing and adult rat, manuscript in prepa
ration.
conserved
Table 2 Clinical profiles of medulloblastoma and PNET
K. Mikoshiba.ThemouseZicgenefamily:homologuesof theDrosophilapairrulegene
@
The
pensable function. Its genomic organization is also highly homolo
gous to that of the mouse.4
Recently, the Drosophila pair-rule gene Opa, which is highly
homologous (over 70%) to Zic in the zinc finger region, was isolated
(10), suggesting that Zic is a mammalian homologue of Opa.4 Opa is
required for timely activation of wingless and engrailed in the para
segment subdivision of the Drosophila embryo (10). Both Wnt-1 and
En-2, which are mammalian homologues of wingless and engrailed,
respectively, are involved in cerebellar development according to the
of
medulloblastoma cells in the majority of the cases tested showed
positive immunoreactivity of anti-Zic mAb in the nucleus, while there
appeared to be no immunoreactivity in other tumors (Table I). In
medulloblastoma cells, the positive immunoreactivity of anti-Zic mAb
was present predominantly
for Zic protein.
Human Zic Is Highly Homologous to the Mouse Zic Gene. We
isolated cDNA and genomic clones of human Zic. The deduced amino
acid sequence is highly homologous to that of the mouse (99.6%), and
the homology is significantly higher than that in other developmen
tally regulated genes,such as that between human GLI and mouse GLJ
(88.4%; Ref. 15) and that between human En2 and mouse En2 (90%;
Ref. 29). Our previous Southern blot analyses showed that there are
homologues of mouse Zic in various vertebrates (carp, frogs, lizards,
chicks, and humans; data not shown). These findings suggest that Zic
Miscellaneoustumors
Carcinoid tumors
Malignant lymphoma
Immature teratoma
Malignant Schwannoma
Alveolar soft part sarcoma
Yolk sac tumor
Liposarcoma
Giant cell sarcoma
negative
DISCUSSION
0/6
0/5
0/4
0/6
0/5
Colorectal cancer
Livercancer
Lung cancer
Breast cancer
2) were
unrelated to clinicopathological features such as location, age, and
stages, or no particular histopathological characteristics were identi
fled in these cases. Ewing sarcoma and PNETs in s.c. region also
exhibited negative reactivity, although histological photographs re
vealed that they were small round cell tumors indistinguishable from
medulloblastoma. Zic protein was unambiguously absent in the rather
common epithelial neoplasms and common brain tumors such as
glioma(Fig.3).
Casea
Histological diagnosis
IN VARIOUS TUMORS
1 yr 8mo
4
yr
8 yr 3mo
4
yr
3
yr
13
yr
2 yr Smo
1 yr 6mo
1 yr 4mo
6 yr 3mo
5
yr
3
yr
4
yr
2
yr
6
yr
2 1 yr
48 yr
35
yr
40
yrM―
10
15
PNET
S
a M, male; F, female.
380
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HUMAN Zic cDNA CLONING AND ITS EXPRESSION IN v@aious
TUMORS
d
:a
@
S
•@
@
‘@a:@
:@
•.,
. . ‘*
.
V
$@
@:“-@‘@
Fig. 3. lmmunohistochemical studies of Zic protein in a medulloblastoma and a glioblastoma. a, a 3-year-old male with classical-type medulloblastoma of the cerebellar vermis. H&E
stain.b, anti-Zicimmunohistochemical
stainof thesamecase.Thenucleioftumor cellswith roundnucleiandfine chromatinstructurearestronglystainedin medulloblastomas,
whereas
thecytosolof cellsandadjacent
structures
fail toexpress
immunoreactive
Zicprotein.c,glioblastoma
multiforme
of thecerebral
hemisphere
in a middle-aged
female,whichshows
typical histological features such as pseudopallisading, necrosis, and endothelial proliferation. Photomicrograph displays hypercellular and relative anaplastic parts. H&E stain. d.
anti-Zic immunohistochemical stain in the same case. Note that tumor cells are unambiguously negative in immunoreactivity of Zic protein. X200. Bar, 20 p.m.
results of gene-targeting studies (30—32). Although it is uncertain
whether the hierarchy in Drosophila is conserved in the mammalian
cerebellum, Zic may play important roles in the development of
cerebellum.
Zic Expressionin Normal HumanCerebellum.In humans,the
EGL is formed by a number of cells which migrate through the
marginal layer from the neuroepithelium to the surface of cerebellum
in the 12-week embryo. In the 6th month, the EGL begins to release
granule cells which migrate inward and pass through the differenti
ating Purkinje cell layer to form the IGL. The production and migra
tion of granule cells continues for 1.5 years after birth (33). In our
study, Zic protein was expressed in the nuclei of the EGL component
cells and in those of inward-migrating granule cells and postmigrated
granule cells in the IGL, both in the developing and developed
cerebella. The immunostaining in this study supports an idea that Zic
is persistently expressed in the granule cells and its precursor cells in
various
species.
The results
of this study also suggest
that Zic will be
useful in pathophysiologically detecting the depopulation of granule
cells caused by viral infection and toxicosis at the fetal stage.
Predominant Expression of Zk in a Distinct Type of Human
Tumor, Medulloblastoma. We examined various human tumors im
munohistochemically and demonstrated a highly restricted expression
of Zic protein in medulloblastoma, whereas no specific antigen has
thus far been reported as a marker of medulloblastoma. As previously
stated, our studies show that Zic is expressed abundantly but restrict
edly in the cerebellar granule cell lineage. Although it is uncertain that
medulloblastoma are committed to a course of the cerebellar granular
cell lineage, our observations support the idea that medulloblastoma
exhibits a differentiation characteristic of the cerebellar granule cell
lineage in terms of expression of Zic, and this particular population of
the cells probably represents the origin of medulloblastoma. Some
immunohistochemicalstudies have demonstratedthe predominant
capacity of medulloblastoma cells to differentiate along neuronal cell
lines (34—37)and have shown that medulloblastoma shares the same
immunological features as postmitotic cells forming the EGL of the
cerebellum (38—41).These findings are also compatible with the
results of our immunohistochemical analyses demonstrating the pres
ence of Zic protein in medulloblastoma.
381
Downloaded from cancerres.aacrjournals.org on August 3, 2017. © 1996 American Association for Cancer Research.
HUMAN Zic cDNA CLONING AND ITS EXPRESSION IN VARIOUS TUMORS
Concerning the histogenesis of medulloblastoma, there has been an
argument about whether the entity of medulloblastoma is unique in
cerebellar tumor or a more ubiquitous tumor derived from younger
neuroblasts
distributed
throughout
the central
nervous
system
(36,
42—47).The recent concept on the tumor entity of the PNET has
complicated the issue. According to the new edition of the WHO book
on “Histological
Typing of the Central Nervous System,―
the diag
nosis of PNET
is recommended
only as a generic
term for cerebellar
medulloblastoma and neoplasms that are histologically indistinguish
able from medulloblastoma but located in the central nervous system
at sites other than the cerebellum (8). However, although PNETs,
regardlessof their locations, appearto be histologically identical, their
biological behavior may vary considerably with location; patients
with supratentorial PNETs have much worse prognoses than patients
with medulloblastoma, despite similar treatments (17). Furthermore, it
was shown that medulloblastoma has a different cytogenetical profile
from other PNETs (22). Our findings suggest that medulloblastoma
differs from other PNETs or Ewing sarcoma of a more undifferenti
ated origin in terms of expression of Zic. It is possible that medullo
blastoma should be classified into a distinct tumor entity which would
differentiate it from other PNETs. Since few biomarkers have been
known for medulloblastoma, the potentiality of this protein is highly
expected for differential diagnosis of medulloblastoma from other
brain tumors, especially undifferentiated neuroectodermal tumors.
Further characterization of the variety of human tumors, especially
central
and
peripheral
PNETs,
by immunohistochemical
analyses
using the anti-Zic mAb are needed to establish its diagnostic signif
icance.
ACKNOWLEDGMENTS
We thankH. Igarashi(1stDepartmentof Pathology,HamamatsuUniversity
Schoolof Medicine)for technicalassistance;M. Fukuda,M. Hashimoto,and
other members of Mikoshiba's laboratory for valuable discussions; Dr. A.
Miyawaki
(Institute
ing adult cerebellar
of Medical
Science, The University
tissues; Dr. K. Sato (University
of Tokyo)
for provid
of Okayama)
for patho
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Predominant Expression of Human Zic in Cerebellar Granule
Cell Lineage and Medulloblastoma
Naoki Yokota, Jun Aruga, Setsuo Takai, et al.
Cancer Res 1996;56:377-383.
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