Development of molecular oncohematology in Ukraine
Disruption of the genetic component of cells are mandatory element of malignant transformation. For the majority of blood neoplasias genetic disorders have been discovered, and they can be used for diagnosis and appropriate therapy. The data obtained by authors about the role of domains of Bcr-Abl p...
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Цитувати: | Development of molecular oncohematology in Ukraine / G.D. Telegeev, M.V. Dybkov, A.N. Dubrovska, D.A. Miroshnichenko, A.P. Tyutyunnykova, S.S. Maliuta // Вiopolymers and Cell. — 2013. — Т. 29, №. 4. — С. 277-282. — Бібліогр.: 31 назв. — англ. |
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irk-123456789-1526032019-06-13T01:25:25Z Development of molecular oncohematology in Ukraine Telegeev, G.D. Dybkov, M.V. Dubrovska, A.N. Miroshnichenko, D.A. Tyutyunnykova, A.P. Maliuta, S.S. Reviews Disruption of the genetic component of cells are mandatory element of malignant transformation. For the majority of blood neoplasias genetic disorders have been discovered, and they can be used for diagnosis and appropriate therapy. The data obtained by authors about the role of domains of Bcr-Abl protein (the main etiological factor in the pathogenesis of leukemia with Ph-chromosome) are presented in this review as well as approved diagnostic methods for myeloproliferative disorders and acute leukemias. Обов’язковим і характерним елементом злоякісної трансформації є порушення генетичного компонента клітини. Для більшості неоплазій системи крові генетичні порушення є відомими, що дозволяє використовувати їх для діагностики і відповідної терапії. Наведено авторські дані стосовно ролі доменів білка Bcr-Abl (головного етіологічного фактора в патогенезі лейкемій з філадельфійською хромосомою) та представлено апробовані методи діагностики мієлопроліферативних захворювань і гострих лейкемій. Обязательным и характерным элементом злокачественной трансформации являются нарушения генетического компонента клетки. Для большинства неоплазий системы крови известны генетические нарушения, что позволяет использовать их для диагностики и проведения соответствующей терапии. Приведены авторские данные о роли доменов белка Bcr-Abl (главного этиологического фактора в патогенезе лейкемий с филадельфийской хромосомой) и представлены апробированные методы диагностики миелопролиферативных заболеваний и острых лейкемий. 2013 Article Development of molecular oncohematology in Ukraine / G.D. Telegeev, M.V. Dybkov, A.N. Dubrovska, D.A. Miroshnichenko, A.P. Tyutyunnykova, S.S. Maliuta // Вiopolymers and Cell. — 2013. — Т. 29, №. 4. — С. 277-282. — Бібліогр.: 31 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000822 http://dspace.nbuv.gov.ua/handle/123456789/152603 577.2.575 en Вiopolymers and Cell Інститут молекулярної біології і генетики НАН України |
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Reviews Reviews Telegeev, G.D. Dybkov, M.V. Dubrovska, A.N. Miroshnichenko, D.A. Tyutyunnykova, A.P. Maliuta, S.S. Development of molecular oncohematology in Ukraine Вiopolymers and Cell |
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Disruption of the genetic component of cells are mandatory element of malignant transformation. For the majority of blood neoplasias genetic disorders have been discovered, and they can be used for diagnosis and appropriate therapy. The data obtained by authors about the role of domains of Bcr-Abl protein (the main etiological factor in the pathogenesis of leukemia with Ph-chromosome) are presented in this review as well as approved diagnostic methods for myeloproliferative disorders and acute leukemias. |
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Telegeev, G.D. Dybkov, M.V. Dubrovska, A.N. Miroshnichenko, D.A. Tyutyunnykova, A.P. Maliuta, S.S. |
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Telegeev, G.D. Dybkov, M.V. Dubrovska, A.N. Miroshnichenko, D.A. Tyutyunnykova, A.P. Maliuta, S.S. |
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Telegeev, G.D. |
title |
Development of molecular oncohematology in Ukraine |
title_short |
Development of molecular oncohematology in Ukraine |
title_full |
Development of molecular oncohematology in Ukraine |
title_fullStr |
Development of molecular oncohematology in Ukraine |
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Development of molecular oncohematology in Ukraine |
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development of molecular oncohematology in ukraine |
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Інститут молекулярної біології і генетики НАН України |
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2013 |
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citation_txt |
Development of molecular oncohematology in Ukraine / G.D. Telegeev, M.V. Dybkov, A.N. Dubrovska, D.A. Miroshnichenko, A.P. Tyutyunnykova, S.S. Maliuta // Вiopolymers and Cell. — 2013. — Т. 29, №. 4. — С. 277-282. — Бібліогр.: 31 назв. — англ. |
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Вiopolymers and Cell |
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2025-07-14T04:03:42Z |
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fulltext |
UDC 577.2.575
Development of molecular oncohematology in Ukraine
G. D. Telegeev, M. V. Dybkov, A. N. Dubrovska, D. A. Miroshnichenko,
A. P. Tyutyunnykova, S. S. Maliuta
Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnogo Str., Ukraine, 03680
g.d.telegeev@imbg.org.ua
Disruption of the genetic component of cells are mandatory element of malignant transformation. For the majo-
rity of blood neoplasias genetic disorders have been discovered, and they can be used for diagnosis and appro-
priate therapy. The data obtained by authors about the role of domains of Bcr-Abl protein (the main etiological
factor in the pathogenesis of leukemia with Ph-chromosome) are presented in this review as well as approved diag-
nostic methods for myeloproliferative disorders and acute leukemias.
Keywords: myeloproliferative neoplasms, Bcr-Abl, Jak2, molecular pathogenesis, diagnostics.
Introduction. Malignant tumors are the second (after
cardiovascular diseases) cause of deaths in developed
countries. According to the Bulletin of National Cancer
Registry of Ukraine [1], blood system neoplasms were
4.87 % of total cancer cases in 2011. Proportion of deaths
from leukemias is 5.38 % of all cancer deaths cases.
Each year about 7,500–8,000 cases of different blood
neoplasms are recorded. Myeloproliferative diseases
(neoplasms) (MPN) are a biologically heterogenous
group of hematopoietic system diseases, the main featu-
re of which is uncontrolled proliferation of stem hema-
topoietic cells or myelopoiesis progenitor cells [2, 3]. The
MPN part in total blood neoplasms is about 30–35 %.
According to the WHO classification, there are cronic
myeloid leukemia (CML), chronic neutrofilic leukemia,
chronic eosinophilic leukemia, polycythemia vera, es-
sential polycythemia, primary myelofibrosis, and un-
classified myeloproliferative neoplasms [4, 5].
Sharing common morpho-clinical and hematologic
features, these diseases, however, have different mole-
cular basis. One of the most represented diseases in this
group is CML. CML has a distinctive marker – Phila-
delphia (Ph') chromosome, which is a result of recipro-
cal translocation t(9;22) (q34; q11). This chromosome is
present in CML and in some patients with acute lym-
phoblastic leukemia (ALL). A fusion of two genes occurs
at the molecular level: abl on chromosome 9 and bcr on
chromosome 22. The breaks in abl occur mostly bet-
ween 1st and 2nd exones and rarely between 2nd and 3rd.
In contrast, bcr gene has three areas where the breaks
happen frequently. Therefore, depending on the break-
point in bcr gene there are three different forms of Bcr-
Abl protein: p190, p210, p230. Each form was found in
the certain type of leukemia: p210 Bcr-Abl – in 90–95 %
of CML cases, p190 Bcr-Abl – mainly in ALL in adults
(35 % of all ALL cases), while p230 is found in rela-
tively benign neutrophilic form of myeloid leukemia.
The impetus for the studying of molecular and ge-
netic aspects of these diseases, which has been started
at our institute in the 1990s, was a pure practical prob-
lem – the development of a system for detecting chime-
ric fusion of bcr-abl gene in the patients who were diag-
nosed and treated in the hematology departments of Ky-
iv hospitals. The cytogenetic methods used to confirm
the presence of the Philadelphia chromosome required
the bone marrow puncture, a long time for analysis, high-
ly qualified staff, etc. Using the reverse transcriptase po-
lymerase chain reaction with specific primers allowed
277
ISSN 0233–7657. Biopolymers and Cell. 2013. Vol. 29. N 4. P. 277–282 doi: 10.7124/bc.000822
� Institute of Molecular Biology and Genetics, NAS of Ukraine, 2013
us to obtain the results within one day after taking pati-
ents’ peripheral blood samples [6–8]. For the first time
in Ukraine the guidelines «The monitoring of chronic
myeloid leukemia with molecular methods» [9] have
been created and approved by the Ministry of Health.
This allowed us to introduce molecular genetic appro-
aches to clinical practice.
However, as it often happens, the practice highligh-
ted a wide range of unsolved scientific problems. Of
particular interest was the fact that different forms of
Bcr-Abl protein (especially p190 and p210) are speci-
fic for leukemias, which differ by the nature of clinical
manifestations. The difference between these proteins
is that protein Bcr incorporates DH (Dbl-homologous)
and PH (pleckstrin-homologous) domains in the area
distinguishing the forms of chimeric proteins p190 and
p210 Bcr-Abl. The experimental data could not descri-
be the functional activity of these domains as a part of
Bcr, and in p210 Bcr-Abl. PH domain has not been stu-
died at all. It was therefore decided to investigate the
areas that distinguish these forms of Bcr-Abl protein in
patients with Ph'-positive leukemia at different stages of
the disease. The transformation of cells expressing fu-
sion protein Bcr-Abl, is associated with impaired regu-
lation of tyrosine kinase Abl, namely its elevated levels
in tumor cells, but there are the unidentified factors that
lead to an exacerbation of the chronic form of the dise-
ase, its transition to the first stage of acceleration, and
then to blast crisis. It has been shown that the Abl over-
expression in cells does not lead to the tumor develop-
ment, but rather results in inhibition of cell growth and
apoptosis [10].
Therefore it was assumed that these specific featu-
res of Ph'-positive leukemias are caused by Bcr protein
domains.
Analysis of dbl area of cDNA obtained from patients
with CML, ALL. Based on the assumption that in some
cases in the region of protein p210 Bcr-Abl, which is ab-
sent in protein p190 Bcr-Abl, might occur the changes
that lead to the functional similarity of these chimeric
proteins, we have analyzed the structure of the site in
patients with Ph'-positive leukemia at different stages
of disease. As a result, the deletions of varying length
were detected in the region, corresponding to Dbl do-
main of Bcr protein, in several patients with acute occur-
rence of the disease [11, 12]. In all cases, the deletions
did not lead to disruption of reading frame. Similar
changes were observed in the control group of healthy
donors. Thus, we have shown that in some cases, in
patients with CML and ALL with p210 Bcr-Abl at the
acute stage of the disease there is a loss of sections of
different length in DH region. So far, a number of sub-
microscopic deletions of varying length on chromoso-
me 9
th (abl gene), and on chromosome 22 (bcr gene) ha-
ve been shown. In most cases, these changes reflected
the progression of the disease and were an unfavorable
prognostic factor.
The changes in the area of chimeric bcr gene that
differ from the «classical» have been described by seve-
ral authors – e6a2, e8a2, e13a2, e15a2 [13–16]. In most
cases these changes were detected during disease acce-
leration. These transcripts have arisen as a result of non-
canonical translocations that occurred in the areas other
than the areas of M-, m-and�-Bcr of chimeric gene bcr-
abl. The changes described by us were different. The
main difference was that these changes were not a re-
sult of an abnormal translocation type. The classic type
of translocation was confirmed by RT-PCR at the stage
of selecting samples for analysis.
Disease progression could not be a result of the ef-
fect of only one factor. However, it is likely that the
lack of a domain or molecular changes in the region of
exons 3–14 of chimeric gene bcr-abl (DH and PH do-
mains) changes their functional activity, which leads to
the formation of proteins acting differently. DH domains
encode guanine exchange factors (GEFs) and work as a
link between the surface cell receptors to cytokines,
growth factors, G-protein receptors, adhesion, on one
hand, and activation of Rho-like GTPases, on the other.
Proteins p210 Bcr-Abl and p190 Bcr-Abl interact dif-
ferently with GTPases RhoA, Rac1 and Cdc42 [17],
which results in the implementation of their capacities,
including influence on actin cytoskeleton, the NADPH-
oxidase during ROS formation, cell signaling, etc.
Study on actin cytoskeleton in leukocytes of patients
with Ph'-positive leukemia. The actin-binding domains,
present in the chimeric Bcr-Abl protein, interact with
actin cytoskeleton and cellular localization [18]. To
check the possible influence of these domains we have
studied blood cells of patients with CML, leukemic cell
cultures and leukocytes obtained from healthy donors.
Comparative analysis of the actin cytoskeleton structu-
res in polymorphonuclear leukocytes using FITC-la-
beled falloidin allowed us to distinguish three types of
278
TELEGEEV G. D. ET AL.
cell staining – cortical, diffuse and dot-like structures
formation [19]. It has been shown that K562, U937
cells as well as cells of patients with Ph'-positive leuke-
mia, have mostly cortical cytoskeleton distribution. In
cells from healthy donors cortical cytoskeleton was al-
most non-expressed. In some cases, in addition to cortical
F-actin large dot-like structures were detected in the cells
obtained from patients with CML (blast crisis stage).
According to the literature data [18], changes in the
organization of actin cytoskeleton may be associated
with mutations in the DH region of bcr-abl gene and its
dysfunction that affects Rho GTPases. It is well-known
that DH domains are GEFs for Rho GTPases, causing
actin cytoskeleton organization: the formation of phyllo-
poda, lamellopoda, stress fibers etc. To test this Dbl re-
gion of chimeric Bcr-Abl, the protein of patients with
large punctates was used. The analysis revealed the pre-
sence of point mutations in this region [20]: transver-
sions at position 2127 (T/C) and at position 2449 (C/A).
They lead to the substitution of phenylalanine for leu-
cine at position 547 and threonine for lysine at position
654. The multiple alignment analysis of DH domains
from eukaryotic proteins showed that the replacement
of Rhe547/Leu is neutral and does not affect the cataly-
tic DH domain function because it does not change the
class of amino acid residues and belongs to �-helix
H2A region. The latter is located on the inner side of �-
helical DH domain group that is not involved in the in-
teractions with GDP, GTPases and neighboring domains.
Instead, Thr654 mutation leads to substitution of a neut-
ral amino acid residue for basic residue and belongs to
the most conservative area of DH domain – CR3. Muta-
tion Thr654/Lys was found within the CR3 sequence
that forms a hydrophobic pocket for GTPase binding.
This fact suggests that the appearance of charged ami-
no acids in this region may have a decisive influence on
the biological activity of Dbl-homologous domain.
Studying the GEF-activity of Bcr DH domain in vitro
and in vivo. Currently more than 70 known proteins have
DH domain in their structure. Most of them are GEF-fac-
tors for different GTPases [21]. Therefore we have tested
the GEF activity in vitro by radioactive GDP release and
the incorporation of �-S35-GT recombinant proteins
which contained DH and DHPH domains, and GTPa-
ses RhoA, Rac1, Cdc42. The experiments with trans-
formation of 293T cells with DH and DHPH recombinant
constructs and GTPases RhoA, RhoB, Rac1, Rac3,
Cdc42, Wrch1 have been also conducted. In both systems,
no GEF activity on GTPases has been found [22].
Multiple alignment of amino acid sequences of DH
family members showed that DH domain of Bcr has
conserved amino acids that are responsible for forming
the tertiary structure. It has also three conservative areas
(CR1, CR2 and CR3) which may be involved in interac-
tions with GTPases. Thus, the analysis of the primary
sequence suggests that the DH domain of Bcr has all
the structural elements that are present in the family of
these domains, and therefore has the potential to func-
tion as a GEF-factor, perhaps, it might have specificity
for other members of Rho family or Ras superfamily
GTPases. In particular, these potential partners can be the
following GTPases: Rap1, Rab5, Arf1. They have been se-
lected because of their ability to interact with PLC� (pre-
sence of RA domain), the ratio of PH domain to memb-
rane phospholipids (Rab5) and Golgi apparatus (Arf1).
Study on p190 and p210 Bcr-Abl proteins by fluores-
cent microscopy. As it was mentioned above, the diffe-
rence between p190 and p210 Bcr-Abl is that p190
lacks the DH and PH part. It is known that there is a di-
vision within lipids by cellular compartments, so the
presence of phosphatydylinositol-monophosphate-bin-
ding PH domain and DH domain in protein can most li-
kely affect the localization of proteins and cytoskeletal
organization.
Cos-1 cells were transfected with the constructs ex-
pressing proteins p190 or p210 Bcr-Abl. The difference
in their localization in cells has been shown in [23]. Both
proteins were found in the cytoplasm, but p190 Bcr-Abl
has an equable distribution in the cytoplasm of cell while
protein p210 is characterized by mainly perinuclear dis-
tribution. So, Bcr part which is present in p210 and ab-
sent in p190 Bcr-Abl, has certain influence on the distri-
bution of proteins. It has been supposed that PH domain
takes part in Bcr-Abl localization. Cos-1 cells morpholo-
gy did not change during transfection. Commonly, cells
are round-shaped and have uniform distribution of actin
filaments, cell membranes form neither lamellopoda
nor phylopoda.
The influence of DH and PH domains was also tes-
ted in NIH3T3 cells which have been transfected with
GFP vectors carrying these domains. The absence of ob-
vious morphological changes was confirmed. This may
mean that Rho family GTPases, involved in the reor-
ganization of actin cytoskeleton, in this case were not
279
DEVELOPMENT OF MOLECULAR ONCOHEMATOLOGY IN UKRAINE
activated, which would be expected if the DH domain of
p210 Bcr-Abl protein had GEF-activity.
Determination of lipids interacting with PH domain
of Bcr. PH domain has been described for more than 200
proteins, and it is involved in cell signaling, cell–cell in-
teractions and cytoskeleton organization. It is known that
in most proteins PH domain binds to phosphatydylino-
sitols, so we decided to check if it is true for Bcr-Abl. In-
deed, it was determined that Bcr-Abl binds to PI(3)P,
PI(4)P and PI(5)P phosphatydylinositol-monophospha-
tes [23, 24]. It has been shown that this interaction is
highly affine [24]. The specificity of binding only phos-
phatydylinositol-monophosphate is not typical for PH
family. Usually, PH domains bind with high affinity
three phosphatydylinositol-three- and bisphosphates that
are the products of PI3-kinase. The cellular lipids provide
the distribution of signaling molecules and organization
of compartment – specific signaling complexes [25].
That is, binding of Bcr or p210 Bcr-Abl to PI(3)P
and PI(4)P can be a result of early endosomal or pha-
gosomal or Golgi complex membrane localization of
these proteins. The role of the binding of PH domain to
PIP(5), which is localized predominantly in the nuc-
leus, may be significant.
Investigation and analysis of the ability of Bcr PH do-
main to bind cellular proteins. It is known that PH do-
mains of some proteins in addition to binding to lipids
may be involved in protein-protein interactions [26]. To
test this fact, the «pull-down» approach was used for
the precipitation of proteins from K562 cells lysate and
their subsequent analysis by proteomics methods. As a re-
sult, 26 proteins that interact with this domain were iden-
tified [23]. The next step was an analysis of interactions
with proteins SMC1 (structure maintenance of chromo-
somes), �-tubulin, zizimin 1 and phospholipase C� (PLC
�). To analyze the binding of PH domain, the DHPH con-
struct, containing both PH and DH domains, was used.
As a control, the DH only construct was used. Using a
pull-down analysis, immunoblotting, and co-localiza-
tion assay in HEK293T cells, the binding of PH domain
to all proteins listed above was confirmed [23, 24]. The
proteins identified belong to several functional groups,
most proteins are involved in metabolic processes, cell
proliferation, adhesion, signal transduction.
Using bioinformatic methods, the network of pro-
teins and signaling pathways that interact with the PH
domain has been constructed. It has three nodal centers:
ERK, NF�B and p38MAPK. Interaction with cortactin
and protein FBP17, which are involved in the forma-
tion of early endosoms, may suggest that Bcr and Bcr-
Abl participate in this process. This assertion was sup-
ported by established interaction with Bcr PH domain
of Bcr with PIP(3) – typical endosomal and phagoso-
mal membrane lipids [23] and with ESCRT complex
(proteins TSG-101 and Vps28) [27]. PLC� also partici-
pates in this process [23]. PLC� catalyzes the cleavage
of phosphatydylinositol-4,5-bisphosphate to diacylgly-
cerol and inositol-1,4,5-triphosphate (Ins(1,4,5)P3),
which is a secondary messenger in many signaling path-
ways. PLC� has a bifunctional role: it is involved in
lipid metabolism and also acts as GEF-factor for Ras
GTPases. So, PLC� can regulate Ras (through its GEF-
domain) and bind Ras-GTP, which potentially creates a
complex mechanism of feedback regulation of a multi-
component system.
Localization of p190 and p210 Bcr-Abl in the cell
relative to the Golgi apparatus. As it has been mentio-
ned above we have shown that the PH domain has the
ability to bind PI(4)P, which is the main component of
lipid membranes of Golgi complex. It was therefore
decided to investigate the co-localization of proteins
p190 and p210 Bcr-Abl with Golgi apparatus. To do
this, Cos-1 cells were transfected with the constructs ex-
pressing proteins p190 and p210 Bcr-Abl. For visualiza-
tion of the Golgi apparatus the antibodies to the matrix
protein GM130 were used.
It has been noticed that the protein p210 was co-
localized with the Golgi apparatus, which typical shape
was observed; in addition, p210 was also localized in
cytoplasm. However, the highest concentration of pro-
tein was observed in the Golgi complex. p190 Bcr-Abl
did not have such obvious location around the nucleus,
it was evenly distributed throughout the cytoplasm of
cells [23]. The specificity of the interaction was tested
by adding wortmannin which led to delocalization of
p210 Bcr-Abl in the area where it has been detected
before treatment. The results of co-transfection with
costructs bearing shRNA specific to PI4K and PTEN al-
so showed the effect of reduced binding of p210 to Gol-
gi complex.
Thus, it was shown that the presence of PH domain
affects the subcellular localization of this protein, cau-
ses the interaction with various proteins and organelles
of the cell. First, it is a participation in the endosomes
280
TELEGEEV G. D. ET AL.
formation. The proteins that interact with the PH-do-
main of Bcr (cortactin, FBP 17, PLC�, �-tubulin) take
part in this process. Thus, the interaction of p210 Bcr-
Abl protein with TSG10, a component of the complex
ESCRT1, might be explained [27]. Despite the lack of
C2 domain of Bcr protein, which is responsible for Bcr
binding to the protein TSG101, the interaction is still
observed in the cells expressing p210 Bcr-Abl. This fact
is well explained by the data established by binding
PH domain to PI(3)P, a dominant early endosomal li-
pid. The absence of this domain in protein p190 Bcr-
Abl alters endosomal sorting and direction of growth
factors receptors transport and differentiation. Differen-
tiation of hematopoietic cells depends not only on the
growth factors receptors, but also on their internaliza-
tion and degradation in time. Thus, aggravation of en-
dosomal transport and accumulation of differentiation
factors, which cannot be degraded, on the cell surface
may affect the differentiation of hematopoietic cells.
The presence of PH domain determines the high affini-
ty of binding Bcr to PI(3)P on early endosomes and pha-
gosomes. Lack of GAP domain at COOH end of Bcr
part leads to deregulation of Rac GTPase, resulting in
the constitutive activity of Rac in hematopoietic cells,
increased formation of reactive oxygen species and
emerging of «oxidative burst» [28].
Improving the detection of Philadelphia chromosome
in patients with CML and ALL and monitoring patients
during treatment with Abl kinase inhibitors. One of the
main problems in the treatment of CML is emerging of
new mutations that make the drugs, including Abl kina-
se inhibitors, unable to work. More than 50 mutations
within the kinase domain of Bcr-Abl protein, most of
which lead to lost or reduced efficiency of Imatinib, one
of the main drugs for CML treatment, have been descri-
bed. Therefore, detection of these mutations is impor-
tant for selecting the treatment strategy. Since most of
these mutations arise in region 200– 400AK of Abl, and
mutations M244V, G250E, Y253F/H, E255K/V, T315I,
M351T, F359V are more than 85 % of the mutational
changes, the primers for nested PCR of this site were
designed. The most common mutation is T315I, which
leads to the replacement of threonine by isoleucine and
causes a loss of effectiveness of the imatinib [29].
Thus, in this study, it has been shown for the first ti-
me that Bcr part of the Bcr-Abl fusion protein plays a
significant role in the mechanisms of tumor develop-
ment during CML progression. It determines the speci-
ficity of different oncoproteins (p190, p210, p230), due
to their different localization in the cell and participa-
tion in different signaling cascades. This fact, in addi-
tion to the traditional use of Abl-kinase inhibitors, in-
creases the possibility of targeted therapy of Ph'-positi-
ve leukemia. Given the high genetic instability in pati-
ents with myeloproliferative disorders, the test systems
for a) detection and differential diagnosis of Ph'-positi-
ve leukemias by RT-PCR, and b) monitoring changes in
ATP-binding region of Abl protein were developed, al-
lowing clinicians to adjust therapy when mutations in
this region occure during tyrosine kinase inhibitor
treatment. Under the current classification of the World
Health Organization (4.5) molecular diagnostics is an
obligatory component of modern diagnostics of leuke-
mias. In collaboration with the colleagues from the Im-
munocytochemistry department of Kavetsky Institute
of Experimental Pathology, Oncology and Radiobiolo-
gy of NAS of Ukraine) we have developed a protocol
for detection of the most characteristic MPN mutations,
namely Jak2, mpl, tet2, asxl1 [30], and mutations in
acute leukemia mll/af4 (t (4, 11) (q21; q23); mll/af9 (t
(9; 11) (p22; q23); cbfb/myh11 (inv16) [31]. The effec-
tiveness of this protocol was tested on a large number of
patients. This greatly improved diagnostic and therapeu-
tic protocols for leukemias of various origins in Ukraine.
Ã. Ä. Òåëåãåºâ, Ì. Â. Äèáêîâ, À. Ì. Äóáðîâñüêà, Ä. Î. ̳ðîøíè÷åíêî,
À. Ï. Òþòþííèêîâà, Ñ. Ñ. Ìàëþòà
Ðîçâèòîê ìîëåêóëÿðíî¿ îíêîãåìàòîëî㳿 â Óêðà¿í³
Ðåçþìå
Îáîâ’ÿçêîâèì ³ õàðàêòåðíèì åëåìåíòîì çëîÿê³ñíî¿ òðàíñôîðìà-
ö³¿ º ïîðóøåííÿ ãåíåòè÷íîãî êîìïîíåíòà êë³òèíè. Äëÿ á³ëüøîñò³
íåîïëàç³é ñèñòåìè êðîâ³ ãåíåòè÷í³ ïîðóøåííÿ º â³äîìèìè, ùî äîç-
âîëÿº âèêîðèñòîâóâàòè ¿õ äëÿ ä³àãíîñòèêè ³ â³äïîâ³äíî¿ òåðàﳿ.
Íàâåäåíî àâòîðñüê³ äàí³ ñòîñîâíî ðîë³ äîìåí³â á³ëêà Bcr-Abl (ãî-
ëîâíîãî åò³îëîã³÷íîãî ôàêòîðà â ïàòîãåíåç³ ëåéêåì³é ç ô³ëàäåëü-
ô³éñüêîþ õðîìîñîìîþ) òà ïðåäñòàâëåíî àïðîáîâàí³ ìåòîäè ä³àã-
íîñòèêè 쳺ëîïðîë³ôåðàòèâíèõ çàõâîðþâàíü ³ ãîñòðèõ ëåéêåì³é.
Êëþ÷îâ³ ñëîâà: 쳺ëîïðîë³ôåðàòèâí³ íåîïëàçìè, Bcr-Abl, Jak2,
ìîëåêóëÿðíèé ïàòîãåíåç, ä³àãíîñòèêà.
Ã. Ä. Òåëåãååâ, Ì. Â. Äûáêîâ, À. Í. Äóáðîâñêàÿ, Ä. À. Ìèðîøíè÷åíêî,
À. Ï. Òþòþííèêîâà, Ñ. Ñ. Ìàëþòà
Ðàçâèòèå ìîëåêóëÿðíîé îíêîãåìàòîëîãèè â Óêðàèíå
Ðåçþìå
Îáÿçàòåëüíûì è õàðàêòåðíûì ýëåìåíòîì çëîêà÷åñòâåííîé
òðàíñôîðìàöèè ÿâëÿþòñÿ íàðóøåíèÿ ãåíåòè÷åñêîãî êîìïîíåí-
281
DEVELOPMENT OF MOLECULAR ONCOHEMATOLOGY IN UKRAINE
òà êëåòêè. Äëÿ áîëüøèíñòâà íåîïëàçèé ñèñòåìû êðîâè èçâåñòíû
ãåíåòè÷åñêèå íàðóøåíèÿ, ÷òî ïîçâîëÿåò èñïîëüçîâàòü èõ äëÿ äè-
àãíîñòèêè è ïðîâåäåíèÿ ñîîòâåòñòâóþùåé òåðàïèè. Ïðèâåäåíû
àâòîðñêèå äàííûå î ðîëè äîìåíîâ áåëêà Bcr-Abl (ãëàâíîãî ýòèî-
ëîãè÷åñêîãî ôàêòîðà â ïàòîãåíåçå ëåéêåìèé ñ ôèëàäåëüôèéñêîé
õðîìîñîìîé) è ïðåäñòàâëåíû àïðîáèðîâàííûå ìåòîäû äèàãíî-
ñòèêè ìèåëîïðîëèôåðàòèâíûõ çàáîëåâàíèé è îñòðûõ ëåéêåìèé.
Êëþ÷åâûå ñëîâà: ìèåëîïðîëèôåðàòèâíûå íåîïëàçìû, Bcr-Abl,
Jak2, ìîëåêóëÿðíûé ïàòîãåíåç, äèàãíîñòèêà.
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