Identification of tumor-associated antigens in human thyroid papillar carcinoma
In this study two cDNA expressing libraries generated from thyroid papillar carcinomas were screened using SEREX approach. Thirty positive cDNA clones representing seventeen different genes were identified from both libraries. It is important to note, that three of them were isolated previously by o...
Gespeichert in:
| Datum: | 2003 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | English |
| Veröffentlicht: |
Інститут молекулярної біології і генетики НАН України
2003
|
| Schriftenreihe: | Біополімери і клітина |
| Schlagworte: | |
| Online Zugang: | https://nasplib.isofts.kiev.ua/handle/123456789/156694 |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Zitieren: | Identification of tumor-associated antigens in human thyroid papillar carcinoma / N.V. Rodnin, I.O. Tykhonkova, R.G. Kyyamova, O.M. Garifulin, I.T. Gout, V.V. Filonenko // Вiopolymers and Cell. — 2003. — Т. 19, № 6. — С. 541-547. — Бібліогр.: 23 назв. — англ. |
Institution
Digital Library of Periodicals of National Academy of Sciences of Ukraine| id |
nasplib_isofts_kiev_ua-123456789-156694 |
|---|---|
| record_format |
dspace |
| spelling |
nasplib_isofts_kiev_ua-123456789-1566942025-02-23T17:28:39Z Identification of tumor-associated antigens in human thyroid papillar carcinoma Ідентифікація пухлиноасоційованих антигенів папілярної карциноми щитовидної залози людини Идентификация опухолеассоциированных антигенов папиллярной карциномы щитовидной железы человека Rodnin, N.V. Tykhonkova, I.O. Kyyamova, R.G. Garifulin, O.M. Gout, I.T. Filonenko, V.V. Біомедицина In this study two cDNA expressing libraries generated from thyroid papillar carcinomas were screened using SEREX approach. Thirty positive cDNA clones representing seventeen different genes were identified from both libraries. It is important to note, that three of them were isolated previously by other laboratories in SEREX screens of various types of human cancer. These include transcription factor NZF, a-catenin and BAG RP11 — a protein with unknown function. Moreover, we identified a whole panel of novel potential tumor-associated antigens, which would be further investigated. We are particularly interested in more detailed analysis of cathepsin H and transducer of ErbB2 (TOB2), which are differentially expressed in various types of human cancer. We will analyse the frequency of autoantibodies against identified antigens in sera of patients with various malignancies and healthy donors by heterologous screening. It is expected that among the clones isolated in this study, there might be novel cancer-associated markers. З двох зразків тканини папіломи щитовидної залози людини отримано дві кДНК експресуючі бібліотеки. Імуноскринуванням бібліотек методом SEREX ідентифіковано 30 позитивних клонів, які відповідали 17 різним генам. Потрібно відмітити, що три гени – транскрипційний фактор NZF, α-катенін і білок ВАС RP11 з поки невідомою функцією – раніше виявлено в інших лабораторіях методом SEREX, де скрикували бібліотеки з різних типів пухлин людини. Серед решти ідентифікованих генів найцікавішими є катепсин Н (cathepsin Н) и ТОВ2 (transducer of ErbB2), підвищену експресію яких було знайдено в багатьох злоякісних пухлинах людини. Подальші дослідження буде направлено на виявлення частоти зустрічальності антитіл проти даних антигенів у сироватках крові хворих на рак різної етіології та здорових донорів. Серед визначених у нашій лабораторії нових SEREX позитивних клонів є потенційні маркери злоякісних новоутворень щитовидної залози людини. кДНК экспрессирующие библиотеки получены из двух образцов ткани злокачественной папилломы щитовидной железы человека. Иммуноскрининг библиотек методом SEREX дал возможность идентифицировать 30 положительных клонов, представляющих собой продукты 17 различных генов. Следует отметить, что три гена — транскрипционный фактор NZF, а-катенин и белок с неизвестной функцией ВАС RP11 – выявлены ранее в других лабораториях SEREX скринингом библиотек из различных типов опухолей человека. Среди остальных идентифицированных генов наибольший интерес представляют катепсин Н (cathepsin Н) и ТОВ2 (transducer of ErbB2), повышенная экспресия которых выявлена во многих злокачественных опухолях человека. Дальнейшие исследования будут направлены на установление частоты встречаемости антител против данных антигенов в сыворотках крови больных раком различной этиологии и здоровых доноров. Среди выявленных нами новых SEREX положительных клонов могут быть потенциальные маркеры злокачественных новообразований щитовидной железы человека. 2003 Article Identification of tumor-associated antigens in human thyroid papillar carcinoma / N.V. Rodnin, I.O. Tykhonkova, R.G. Kyyamova, O.M. Garifulin, I.T. Gout, V.V. Filonenko // Вiopolymers and Cell. — 2003. — Т. 19, № 6. — С. 541-547. — Бібліогр.: 23 назв. — англ. 0233-7657 DOI:http://dx.doi.org/10.7124/bc.000681 https://nasplib.isofts.kiev.ua/handle/123456789/156694 577.29:577.27 en Біополімери і клітина application/pdf Інститут молекулярної біології і генетики НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Біомедицина Біомедицина |
| spellingShingle |
Біомедицина Біомедицина Rodnin, N.V. Tykhonkova, I.O. Kyyamova, R.G. Garifulin, O.M. Gout, I.T. Filonenko, V.V. Identification of tumor-associated antigens in human thyroid papillar carcinoma Біополімери і клітина |
| description |
In this study two cDNA expressing libraries generated from thyroid papillar carcinomas were screened using SEREX approach. Thirty positive cDNA clones representing seventeen different genes were identified from both libraries. It is important to note, that three of them were isolated previously by other laboratories in SEREX screens of various types of human cancer. These include transcription factor NZF, a-catenin and BAG RP11 — a protein with unknown function. Moreover, we identified a whole panel of novel potential tumor-associated antigens, which would be further investigated. We are particularly interested in more detailed analysis of cathepsin H and transducer of ErbB2 (TOB2), which are differentially expressed in various types of human cancer. We will analyse the frequency of autoantibodies against identified antigens in sera of patients with various malignancies and healthy donors by heterologous screening. It is expected that among the clones isolated in this study, there might be novel cancer-associated markers. |
| format |
Article |
| author |
Rodnin, N.V. Tykhonkova, I.O. Kyyamova, R.G. Garifulin, O.M. Gout, I.T. Filonenko, V.V. |
| author_facet |
Rodnin, N.V. Tykhonkova, I.O. Kyyamova, R.G. Garifulin, O.M. Gout, I.T. Filonenko, V.V. |
| author_sort |
Rodnin, N.V. |
| title |
Identification of tumor-associated antigens in human thyroid papillar carcinoma |
| title_short |
Identification of tumor-associated antigens in human thyroid papillar carcinoma |
| title_full |
Identification of tumor-associated antigens in human thyroid papillar carcinoma |
| title_fullStr |
Identification of tumor-associated antigens in human thyroid papillar carcinoma |
| title_full_unstemmed |
Identification of tumor-associated antigens in human thyroid papillar carcinoma |
| title_sort |
identification of tumor-associated antigens in human thyroid papillar carcinoma |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2003 |
| topic_facet |
Біомедицина |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/156694 |
| citation_txt |
Identification of tumor-associated antigens in human thyroid papillar carcinoma
/ N.V. Rodnin, I.O. Tykhonkova, R.G. Kyyamova, O.M. Garifulin, I.T. Gout, V.V. Filonenko // Вiopolymers and Cell. — 2003. — Т. 19, № 6. — С.
541-547. — Бібліогр.: 23 назв. — англ. |
| series |
Біополімери і клітина |
| work_keys_str_mv |
AT rodninnv identificationoftumorassociatedantigensinhumanthyroidpapillarcarcinoma AT tykhonkovaio identificationoftumorassociatedantigensinhumanthyroidpapillarcarcinoma AT kyyamovarg identificationoftumorassociatedantigensinhumanthyroidpapillarcarcinoma AT garifulinom identificationoftumorassociatedantigensinhumanthyroidpapillarcarcinoma AT goutit identificationoftumorassociatedantigensinhumanthyroidpapillarcarcinoma AT filonenkovv identificationoftumorassociatedantigensinhumanthyroidpapillarcarcinoma AT rodninnv ídentifíkacíâpuhlinoasocíjovanihantigenívpapílârnoíkarcinomiŝitovidnoízalozilûdini AT tykhonkovaio ídentifíkacíâpuhlinoasocíjovanihantigenívpapílârnoíkarcinomiŝitovidnoízalozilûdini AT kyyamovarg ídentifíkacíâpuhlinoasocíjovanihantigenívpapílârnoíkarcinomiŝitovidnoízalozilûdini AT garifulinom ídentifíkacíâpuhlinoasocíjovanihantigenívpapílârnoíkarcinomiŝitovidnoízalozilûdini AT goutit ídentifíkacíâpuhlinoasocíjovanihantigenívpapílârnoíkarcinomiŝitovidnoízalozilûdini AT filonenkovv ídentifíkacíâpuhlinoasocíjovanihantigenívpapílârnoíkarcinomiŝitovidnoízalozilûdini AT rodninnv identifikaciâopuholeassociirovannyhantigenovpapillârnojkarcinomyŝitovidnojželezyčeloveka AT tykhonkovaio identifikaciâopuholeassociirovannyhantigenovpapillârnojkarcinomyŝitovidnojželezyčeloveka AT kyyamovarg identifikaciâopuholeassociirovannyhantigenovpapillârnojkarcinomyŝitovidnojželezyčeloveka AT garifulinom identifikaciâopuholeassociirovannyhantigenovpapillârnojkarcinomyŝitovidnojželezyčeloveka AT goutit identifikaciâopuholeassociirovannyhantigenovpapillârnojkarcinomyŝitovidnojželezyčeloveka AT filonenkovv identifikaciâopuholeassociirovannyhantigenovpapillârnojkarcinomyŝitovidnojželezyčeloveka |
| first_indexed |
2025-11-24T02:43:35Z |
| last_indexed |
2025-11-24T02:43:35Z |
| _version_ |
1849637957837783040 |
| fulltext |
ISSN 0233-7657. Біополімери і клітина. 2003. Т. 19. № 6
Identification of tumor-associated antigens in human
thyroid papillar carcinoma
N. V. Rodnin 1, I. O. Tykhonkova 1, R. G- Kyyamova 1 , О. M. Garifulin1, I. T. Gout 1 2 ,
V. V. Filonenko 1
1 Institute of Molecular Biology and Genetics NAS of Ukraine
150 vul. Acad. Zabolotnoho, Kyiv 03143, Ukraine
2
Ludwig Institute for Cancer Research
91 Riding House Street, London W1W 7BT, UK
In this study two cDNA expressing libraries generated from thyroid papillar carcinomas were screened
using SEREX approach. Thirty positive cDNA clones representing seventeen different genes were identified
from both libraries. It is important to note, that three of them were isolated previously by other laboratories
in SEREX screens of various types of human cancer. These include transcription factor NZF, a-catenin
and ВАС RPU—a protein with unknown function. Moreover, we identified a whole panel of novel
potential tumor-associated antigens, which would be further investigated. We are particularly interested in
more detailed analysis of cathepsin H and transducer of ErbB2 (TOB2), which are differentially expressed
in various types of human cancer. We will analyse the frequency of autoantibodies against identified
antigens in sera of patients with various malignancies and healthy donors by heterologous screening. It is
expected that among the clones isolated in this study, there might be novel cancer-associated markers.
Introduction. Thyroid cancer, although the most fre
quent malignancy of the endocrine system, is in
general a rare disease. It accounts for about 1 % of
all human cancers, with a higher prevalence in women
(5—9 of 100000) as compared with men (2—4 of
100000) [1]. Normally, thyroid cancer is a disease
with good prognosis, but about 30 % of tumors
dedifferentiate and may finally develop into highly
malignant anaplastic thyroid carcinomas with the
mean survival time of less than 8 months. The thyroid
gland is highly sensitive to radiation-induced on
cogenesis. This is verified by numerous reports from
survivors after Hiroshima and Nagasaki, the Nevada,
Novaya Zemlya and Marshal Island atomic bomb
tests [2]. Investigations provided after Chernobyl
nuclear plant accident have shown the increase of
thyroid cancer two fold in adults and three fold in
children.
The success of cancer therapy depends on the
stage of the disease detection. Today, the specific
markers for early detection of thyroid cancer are
© N. V. RODNIN, I. O. TYKHONKOVA, R. G. KYYAMOVA,
О. M. GARIFULLIN, I. T. G O U T , V. V. FILONENKO, 2 0 0 3
unknown and the search for them is a problem of
outstanding value.
At present, several methods for the search of
tumor-associated antigens exist, such as DNA Micro-
arrays, SAGE, CNAPS, SEREX (Serological analysis
of recombinant cDNA expressing libraries) with their
advantages and deficiencies [3]. SEREX method is
based on the study of cancer patient's immune
response. Samples of cancer tissue are used for the
creation of expressing cDNA libraries further scree
ned by autologous sera. This method is suitable for
the identification of the proteins (or their immuno
genic epitopes) that cause immune responses in can
cer patients.
SEREX method has been successfully applied for
the detection of tumor-associated antigens from vari
ous types of human cancer, including breast, colo
rectal, renal etc. [4—6].
These studies led to the identification of several
novel antigens, which are currently used as markers
of malignant transformation and for the production of
anti-cancer vaccines [7 ]. We have previously reported
541
RODNIN N. V. Б Т AL.
the identification of 15 immunoreactive clones isolated
by SEREX screening of thyroid cancer cDNA exp
ressing libraries [8 ]. We have extended the search for
novel antigens by generating and screening of two
novel libraries from thyroid papillar cancer. Here, we
report the identification of further 30 immunoreactive
clones, representing 17 genes. It is important to note
that three from identified genes have been previously
found by SEREX approach from other types of human
cancer. Heterologous screens of isolated clones with a
panel of sera from healthy donors and patients with
various types of cancer is currently in progress.
Materials and Methods. The samples of human
thyroid papillar carcinomas and autologous sera were
kindly provided by Dr. V. Usenko and Dr. V.
Lysogubov (BIONTEC, Ukraine).
Purification of total and messenger RNA. Total
RNA was purified from freshly frozen in liquid
nitrogen tumor samples by the modified guanidine-
isothiocyanate method [9 J. mRNA was isolated by
affinity chromatography using oligo(dT) Dynabeds
matrix («Dynal», UK). The quality and quantity of
the preparations obtained were estimated by spectro-
photometrical methods and by the electrophoresis in
1 % formaldehyde-agarose gels. The mRNA prepa
rations were stored under ethanol at -80 °С until
further use.
Generation of cDNA expressing libraries. Five
micrograms of purified mRNA preparations were
applied for cDNA synthesis with the use of cDNA
synthesizing kit («Stratagene», USA). The oligonuc
leotide primer containing Xhol site and an oligo(dT)
tail was used for the initiation of reverse transcription.
The effectiveness of the synthesis was estimated with
the use of the test RNA preparation from Stratagene
kit followed by electrophoresis in 1 % agarose gels.
cDNA fragments accounting 0.5—3 kb were
extracted from the gels using DNA extraction kit
(«Qiagen», USA). The fragments obtained were clo
ned into bacteriophage Я DNA (UniZap vector) by
Xhol and EcoRI restriction sites. The phagemide
obtained was packed into phage particles with the use
of Gigapack Gold III kit («Stratagene», USA). For the
estimation of primary libraries titers Escherichia coli
XL-1 Blue MRF' cells were infected by recombinant
phages in appropriate dilutions. The percentage of
non-recombinant phages was detected by blue/white
plaque selection in the presence of IPTG and X-Gal
(«Sigma», USA).
Affinity purification of the sera. Sera samples
from patients with thyroid cancer were diluted 1:1
with glycerol and stored at -20 °С until further use.
For the affinity purification, prepared sera samples
were diluted 1:10 by TBS (10 mM Tris-HCl, pH 8.0,
150 mM NaCl) and incubated with affinity matrixes
containing covalently crosslinked E. coli and Я phage
proteins (Y1090 and BNN97 matrixes respectively).
This technique allowed the elimination of serum
immunoglobulins directed against bacterial and phage
proteins. The depleted sera were further diluted by
TBS 1:100 and stored at 4 °С with the addition of
0.02 % NaN 3.
Immunoscreening of cDNA expression libraries.
For the primary screening 600 /лі of E. coli XL-1 Blue
MRF' cells were infected with 6* 10 s phage particles
and plated onto 150 mm Petri dishes. The expression
of recombinant proteins was induced by the addition
of 1 mM IPTG for 5—7 hours. The transfer of the
proteins to the nitrocellulose membrane Hybond-C
was carried out overnight by standard method.
The identification of IgG expressing clones was
performed by immunoblotting of filters with anti-
human horse radish peroxidase conjugate (dilution
1:2000, «Sigma», USA). Detection of primary posi
tives was carried out by probing filters with the
autologous sera and anti-human alkaline phosphatase
conjugate staining. Positive clones were extracted
from the agar and stored in 0.5 ml of SM buffer with
the addition of 20 ці of chloroform.
The secondary screening of isolated clones was
performed on 90 mm Petri dishes by the same
method. Plasmid DNA from positive clones was
obtained by in vivo recombination in E. coli strain
XL-1 Blue MRF'.
Restriction analysis and sequencing of isolated
clones. The size of cDNA insertions was detected by
restriction analysis with EcoRI and Xhol endonuc-
leases followed by 1 % agarose gel electrophoresis.
Sequencing of the inserts was performed by a stan
dard protocol using the automatic sequencer ABI 373
(Applied Biosystem). The identification of positive
clones and their analysis was performed with a help
of EMBO, GenBank, dBest and SEREX databases.
Results and Discussion. Taking into conside
ration the increase of thyroid cancer frequency and
the absence of specific markers for early diagnostic
purposes we have extended our search for thyroid
cancer associated markers. In this study, we have
used SEREX methodology aimed on the identification
of tumor-associated proteins which induce immune
response in cancer patients. We have successfully
applied SEREX technique in previous studies, which
542
IDENTIFICATION OF T U M O R - A S S O C I A T E D ANTIGENS
led us to the identification of 25 immunoreactive
clones from thyroid cancer and melanoma [8, 10].
In order to extend the search for novel antigens,
we have created two additional cDNA expressing
libraries from thyroid papillar carcinomas with the
titer of 1.05 and 1.4 • 106 respectively. The percentage
of non-recombinant phages was less than 1 % for
both libraries. These tests indicated that both li
braries are suitable for immunoscreening by SEREX
methodology.
The primary immunoscreening with autologous
sera allowed us to isolate 170 primary positives
clones, which showed various degree of immuno-
reactivity. It is important to note that we picked up
clones which exhibited even a very weak immunoreac-
tivity (a borderline with non-specific signal). There
fore, a large number of primary clones were not
confirmed by secondary screenings, leaving only 30
clones as true-positives. cDNA plasmids, correspon
ding to positive clones, were rescued by in vitro
recombination approach. The size of inserts in isola
ted plasmids were determined by restriction analysis
(data not shown). The identification of isolated clones
was performed by sequence analysis followed by
searching of various DNA and protein databases. The
search revealed that 30 clones encode 17 genes (Table
1). Furthermore, 12 of them encode known proteins,
while 5 represent so far genes with unknown func
tions. The sequence of isolated clones and other
relevant information were submitted to the SEREX
database http://www.licr.org/SEREX.html.
The analysis of SEREX database revealed that
three of these genes were identified earlier by other
SEREX laboratories in screens of different types of
human cancer (Table 2). These include a potential
transcription factor NZF, a-catenin and a protein with
unknown function ВАС RP11. Noteworthy, NZF
protein was isolated from libraries generated from
glioma, renal cell carcinoma, ovarian and colon can
cer, teratoma and normal testis. On the other hand,
a-catenin was cloned from melanoma, renal, breast,
colorectal cancer, small cell carcinoma and testis.
Further analysis of other antigens identified in pre
sent study showed their relevance to malignant trans
formation.
For example, a- and /?-catenins are involved in
cadherin-mediated cell-cell adhesion. Disregulation of
cellular functions of both a- and /ї-catenins is asso
ciated with invasive potential of malignant cells [11—
13]. The overexpression of ErbB-2 antigen is ob
served in 30 % of breast cancer and is associated with
poor prognosis. Moreover, ErbB2 has been also found
overexpressed in thyroid and other types of human
cancer [14, 15]. In this study we have isolated the
transducer of ErbB2 (clone Thy28). So far, very little
is known about the function of the transducer of
ErbB2. However, there is no doubt that the function
of this gene in normal and transformed cells needs
further investigation.
Phosphotyrosine independent ligand p62 for the
Lck SH2 domain (Thy36) is a major component of
intracytoplasmic hyaline bodies in hepatocellular car
cinoma cells, while in non-neoplastic liver cells it was
not observed [16]. p62 protein binds ubiquitin and
may act as an adapter linking ubiquitinated proteins
to multienzyme proteosomal complexes to other pro
teins. These features suggest a role for p62 in signal
transduction and possibly also in carcinogenesis.
Elevated expression of proteases is observed in a
variety of tumors. Clone Thy40 encodes a protease —
cathepsin H, whose expression is increased in high-
grade prostatic intraepithelial neoplasia and carci
noma of the prostate [17 ]. Two forms of cathepsin are
known: a full length and a truncated version. Both of
them are enzymatically active, but truncated form has
a reduced lysosomal association when compared with
a full-length cathepsin H. It was suggested that
increased expression of cathepsin H may affect cellu
lar functions especially those which are associated
with tumor progression and metastasis. Short-chain
collagen type VIII (Thy43) was observed throughout
the development of hemangioma in the study of Tan
et al. [18]. It was also detected within mast cells
during early proliferative phase. In several SEREX
studies different ribosomal proteins have been iden
tified as tumor-associated antigens [19, 20]. We have
identified ribosomal protein S24 (Thy32 and 35)
which has not been previously detected in serological
screenings by SEREX or other approaches. Several
clones corresponding to non-sarcomeric myosin light
chain (Thy37, 38 and 42) have been also isolated in
our screen. In our previous article [8] we have
reported SEREX-based detection of CDC-42 binding
protein kinase. Phosphorylation of myosin light chain
by CDC-42 binding protein kinase leads to the
activation of actin-myosin contractility [21 ]. Dis
regulation in actin-myosin contractility, induced by
aberrant signaling may play a role in malignant
invasiveness and metastatic growth [22 ].
The clone KY-Thy29 encodes for the solid tumor
associated protein. Two STAG1/PMEPA1 mRNA
transcripts of approximately 2.7 and 5 kb, with
543
http://www.licr.org/SEREX.html
RODNIN N. V. ET AL.
Table 1
Immunoreactive clones isolated by SEREX screening of thyroid cancer libraries
Group number Clone (SEREX ID) Homology/Identiry
4
5
6
7
8
9
10
11
12
13
14
15
16
17
Cathepsin H (CTSH)
Proteine phosphatase 1, regulatory (inhibitor) subunit 15A
Collagen, type VIII, alpha 2
Clone XXbac-44E15 on chromosome 6
identical coding regions but variant 3' untranslated
regions, were predominantly expressed in normal
prostate tissue and at lower levels in the ovary. The
expression of this gene was upregulated in 87 % of
RCC samples and also was upregulated in stomach
and rectal adenocarcinomas.
In contrast, STAG1/PMEPA1 expression was
barely detectable in leukemia and lymphoma samples.
Analysis of expressed sequence tag databases showed
that STAG1/PMEPA1 also was expressed in pancrea
tic, endometrial, and prostatic adenocarcinomas. The
STAG1/PMEPA1 cDNA encodes a 287-amino-acid
protein containing a putative transmembrane domain
and motifs that suggest that it may bind src homology
3- and tryptophan domain-containing proteins. This
protein shows 67 % identity to the protein encoded
by the chromosome 18 open reading frame 1 gene.
Translation of STAG1/PMEPA1 mRNA in vitro sho
wed two products of 36 and 39 kDa, respectively,
suggesting that translation may initiate at more than
one site. The upregulation of this gene in several solid
tumors indicated that it may play an important role
in tumorigenesis [23]. The link between identified
antigens and various aspects of malignant trans
formation, based on the literature search, is summa
rized in Table 3. Our further investigations will be
544
IDENTIFICATION OF T UM OR-ASSOCI ATED ANTIGENS
Table 2
The summary of serological properties and expression pattern of 3 genes, which were identified earlier in different SEREX screenings
Our clone (SEREX ID) Homology/Identi ty Group mRNA sourse/serum Characteristics
KY-Thy 17 (2313) C2H2 zinc finger University of Mainz
protein (NZF)
KY-Thy22 (2418)
Table 3
A possible link between SEREX-derived antigens from human thyroid carcinoma and malignant transformation
Group number Clones Homology
Possible role in cancer
development and progression
Catenin beta-like 1
Alpha-catenin
Transducer of ErbB2
Sequestesoma 1
Non-sarcomeric myosin, light
polypeptide
Cathepsin H (CTSH)
Proteine phosphatase 1, regulatory
(inhibitor) subunit 15A
Collagen, type VIII, alpha 2
The levels of catenin expression in tumor tissue differs
from the normal ones
a-Catenin meets the criteria of an invasion suppressor
gene
The expression in thyroid carcinomas correlates with the
degree of aggressiveness and differentiation
Major component of intracytoplasmic hyaline bodies in
hepatocellular carcinoma
Involved in invasion and metastasis of pancreatic cancer
Increased expression in high-grade prostatic intra
epithelial neoplasia and carcinoma of the prostate
Involved in down-regulation of signalling pathways,
inducing growth and proliferation
Short-chain collagen is localized extracellulary
throughout the development of hemangioma
545
RODNIN N. V. ET AL.
focused on the elucidation of the role of identified
antigens in cancerogenesis. Heterologous screenings
with a panel of sera from healthy donors and patients
with various types of cancer would allow us to select
those which have the properties of tumor specific
markers or the potential for the development of
anti-cancer vaccines.
Acknowledgements. This study was supported in
part by grants from the National Academy of Sciences
of Ukraine, The Royal Society and INTAS. Authors
are grateful to Dr. Usenko and Dr. Lyzogubov (BIO-
NTEC, Ukraine) for providing the samples of thyroid
cancer and autologous sera.
M. В. Роднін, I. О. Тихонкова, P. Г. Кіямова, О. M. Гарифулін,
І. Т. Гут, В. В. Філоненко
Ідентифікація пухлиноасоційованих антигенів папілярної
карциноми щитовидної залози людини
Резюме
З двох зразків тканини папіломи щитовидної залози людини
отримано дві кДНК експресуючі бібліотеки. Імуноскрину-
ванням бібліотек методом SEREX ідентифіковано ЗО пози
тивних клонів, які відповідали 17 різним генам. Потрібно
відмітити, що три гени — транскрипційний фактор NZF,
а-катенін і білок ВАС RP11 з поки невідомою функцією —
раніше виявлено в інших лабораторіях методом SEREX, де
скрикували бібліотеки з різних типів пухлин людини. Серед
решти ідентифікованих генів найцікавішими є катепсин Н
(cathepsin Н) и ТОВ2 (transducer of ErbB2), підвищену екс
пресію яких було знайдено в багатьох злоякісних пухлинах
людини. Подальші дослідження буде направлено на виявлення
частоти зустрічальності антитіл проти даних антигенів у
сироватках крові хворих на рак різної етіології та здорових
донорів. Серед визначених у нашій лабораторії нових SEREX
позитивних клонів є потенційні маркери злоякісних новоутво
рень щитовидної залози людини.
Н. В. Роднин, И. А. Тихонкова, Р. Г. Киямова,
О. М. Гарифулин, И. Т. Тут, В. В. Филоненко
Идентификация опухолеассоциированных антигенов
папиллярной карциномы щитовидной железы человека
Резюме
кДНК экспрессирующие библиотеки получены из двух образцов
ткани злокачественной папилломы щитовидной железы чело
века. Иммуноскрининг библиотек методом SEREX дал воз
можность идентифицировать 30 положительных клонов, пред
ставляющих собой продукты 17 различных генов. Следует
отметить, что три гена — транскрипционный фактор NZF,
а-катенин и белок с неизвестной функцией ВАС RP11 —
выявлены ранее в других лабораториях SEREX скринингом
библиотек из различных типов опухолей человека. Среди ос
тальных идентифицированных генов наибольший интерес
представляют катепсин Н (cathepsin Н) и ТОВ2 (transducer of
ErbB2), повышенная экспресия которых выявлена во многих
злокачественных опухолях человека. Дальнейшие исследования
будут направлены на установление частоты встречаемости
антител против данных антигенов в сыворотках крови боль
ных раком различной этиологии и здоровых доноров. Среди
выявленных нами новых SEREX положительных клонов могут
быть потенциальные маркеры злокачественных новообразова
ний щитовидной железы человека.
REFERENCES
1. Schmutzler С , Koehrle J. Innovative strategies for the treat
ment of thyroid cancer / / Eur. J. Endocrinol.—2000.—143.—
P. 15—24.
2. Shigenoby N., Nystrom E. Epidemiology and primary preven
tion of thyroid cancer / / Thyroid.—2002.—12.—P. 889—
896.
3. Parmiani G. Tumor immunity as autoimmunity: tumor antigens
include normal self proteins which stimulate anergic peripherial
T cells / / Immunol. Today.—1993.—14.—P. 536—538.
4. Old L. J., Chen Y. T. New paths in human cancer serology / /
J. Exp. Med.—1998.—187.—P. 1163—1167.
5. Jager D., Unkelbach M., Frei C, Bert F, Scanlan M. J.,
Jager E., Old L J., Chen Y. Т., Knuth A. Identification of
tumor-restricted antigens NY-BR-1, SCP-1 , and a new can
cer/testis-like antigen NW-BR-3 by serological screening of a
testicular library with breast cancer serum / / Cancer Im
munol.—2002.—N 28.—P. 2—5.
6. Behrends U., Schneider I., Rossler S., Frauenknecht H.t
Golbeck A., Lechner В., Eigenstetter G., Zobywalski C ,
Muller-Weihrich S., Graubner U., Schmid I., Sackerer D.}
Spath M., Goetz C, Prantl F., As muss H. P., Bise K.,
Mautner J. Novel tumor antigens identified by autologous
antibody screening of childhood medulloblastoma cDNA lib
raries / / Int. J. Cancer.—2003.—106, N 2.—P. 244—251.
7. Scanlan M. J., Jager D. Challenges to the development of
antigen-specific breast cancer vaccines / / Breast Cancer
Res .—2001.—3, N 2.—P. 95—98.
8. Rodnin N. V., Tykhonkova I. O., Nemazany I. O., Gorlova L
M., Komissarenko I. V., Palchevskiy S. S., Kuharenko O. P.,
Drobot L. В., Matsuka G. H., Filonenko V. V., Gout L T.
Serological identification of autoimmune reactive antigens in
human thyroid cancer cells / / Exp. Oncol .—2000.—22,
N 3.—P. 135—138.
9. Chomczynski P., Sacchi N. Single-step method of RNA
isolation by acid guanidinium thiocyanate-phenol-chloroform
extraction / / Anal. Biochem.—1987.—162.—P. 156—159.
10. Rodnin M. V., Tykhonkova 1. O., Filonenko V. V., Drobot L
В., Matsuka G. H., Gout I. T. Search and characterization of
melanoma antigens with the use of serological identification of
antigens by recombinant expression cloning method / / Biopo-
lymers and Cel l .—2000.—16.—P. 339—345.
11. Hajra JC M., Fearon E. R. Cadherin and Catenin alterations
in human cancer / / Genes, Chromosmes and Cancer.—
2002.—34.—P. 255—268.
12. Aiken E. V., Wever O. D., Correia da Rocha A. S., Mar eel M.
Defective E-cadherin/catenin complexes in human cancer / /
Virchows Arch.—2001.—439.—P. 725—751 .
13. Shiozaki H., Iihara K., Oka H., Kadowaki Т., Matsui S.,
Gofuku J., Inoue M., Nagafuchi A., Tsukita S., Mori T.
Immunohistochemical detection of alpha-catenin expression in
human cancers / / Amer. J. Pathol .—1994.—144, N 4 .—
P. 667—674.
14. Soda G., Antonaci A., Bosco D., Nardoni S., Metis M.
Expression of bcl-2, c-erbB-2, p53, and p21 (wafl-cipl)
protein in thyroid carcinomas / / J . Exp. Clin. Cancer Res.—
1999.—18, N 3 .—P. 363—367.
15. Lee M. S.t Igawa Т., Yuan Т. C, Zhang X. Q., Lin F. F, Lin
M. F. ErbB-2 signalling is involved in regulating PSA secretion
in androgen-independent human prostate cancer LNCaP C-81
cells / / Oncogene.—2003.—22, N 5 .—P. 781—796.
546
IDENTIFICATION OF T U M O R - A S S O C I A T E D ANTIGENS
16. Stumptner C , Held #., Fuclisbichler A., Hauser #., Mis-
chinger H. /., Zatlokal K.t Denk H. Analysis of intracytoplas-
mic hyaline bodies in hepatocellular carcinoma / / Amer. J.
Pa thol .—1999.—154, N 6 .—P. 1701—1710.
17. Waghray A., Keppler D., Sloane В. F, Schuger L.} Chen Y.
Q. Analysis of a truncated form of cathepsin H in human
prostate tumor cells / / J. Biol. Chem.—2002.—277, N 13 .—
P. 11533—11538.
18. Tan S. Т., Velickovic M., Ruger B. M., Davis P. F. Cellular
and extracellular markers of hemangioma / / Plast. Reconstr.
Surg .—2000.—106, N 3 .—P. 529—538 .
19. Koroleva E. P., Lagarkova M. A., Mesheryakov A. A , Scanlan
M. J., Old L. J., Nedospasov S. A., Kuprash D. V. Serological
identification of antigens associated with renal cell carcinoma
/ / Russ. J. Immunol .—2002.—7.—P. 3229—3238.
20. Uemura M., Nous о К, Kobayashi Y., Tanaka #., Nakamura
S., Higashi Т., О no Т., Nakayama E., Hanafusa Т., Shiratori
Y. Identification of the antigens predominantly reactive with
serum from patients with hepatocellular carcinoma / / Can
ce r .—2003 .—97, N 10 .—P. 2474—2479.
2 1 . Moncrieff C. L., Bailey M. E.y Morrison TV., Johnson K. J.
Cloning and chromosomal localization of human Cdc42-binding
protein kinase beta / / Genomics .—1999.—57, N 2 — P . 297—
300.
22. Kaneko JC, Satoh K., Masamune A., Satoh A., Shimosegawa
T. Myosin light chain kinase inhibitors can block invasion and
adhesion of human pancreatic cancer cell lines / / Pancreas .—
2002.—24, N 1.—P. 3 4 — 4 1 .
23 . Rae F. JC, Hooper /. D., Nicol D. L., Clements J. A.
Characterization of a novel gene, S T A G 1 / P M E P A 1 , upregu-
lated in renal cell carcinoma and other solid tumors / / Мої.
Carc inog .—2001.—321.—P. 4 4 — 5 3 .
У Д К 577.29:577.27
Надійшла до редакції 12.02.02
547
|