Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells
Background: The mechanisms of drug resistance of cancer have not been yet elucidated in details. Recently, the role of mast cells (MCs) in the development of drug resistance has been brought in the limelight. The aim of the study was to examine the morphological features of doxorubicin (DOX)-resista...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2018
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| Цитувати: | Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells / N.V. Boroday, V.F. Chekhun // Experimental Oncology. — 2018 — Т. 40, № 1. — С. 42-47. — Бібліогр.: 30 назв. — англ. |
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irk-123456789-1392462018-06-20T03:06:44Z Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells Boroday, N.V. Chekhun, V.F. Original contributions Background: The mechanisms of drug resistance of cancer have not been yet elucidated in details. Recently, the role of mast cells (MCs) in the development of drug resistance has been brought in the limelight. The aim of the study was to examine the morphological features of doxorubicin (DOX)-resistant Walker 256 carcinosarcoma and to assess the response of MCs and histamine content in these cells in relation to the development of resistance to DOX as well as in DOX-resistant tumors. Materials and Methods: The DOX resistance was induced by serial passages of Walker 256 carcinosarcoma in rats in the setting of DOX treatment in vivo. MCs in tumors were detected in the sections by staining with Toluidine Blue O. Histamine content in MCs stained with solution of Water Blue-Orcein was assessed by Astaldi semiquantitative method taking into account different staining intensity. Results: Formation of DOX resistance in the course of serial passages of Walker 256 carcinosarcoma was accompanied by the increase in the number of MCs in tumors and histamine content. Nevertheless, in tumors with phenotype of complete DOX resistance the number of histamine-containing MCs decreased to the same level as in tumors of the original strain that are DOX-sensitive. Conclusion: MCs are involved in formation of DOX resistance in Walker 256 carcinosarcoma. 2018 Article Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells / N.V. Boroday, V.F. Chekhun // Experimental Oncology. — 2018 — Т. 40, № 1. — С. 42-47. — Бібліогр.: 30 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/139246 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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DSpace DC |
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English |
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Original contributions Original contributions |
| spellingShingle |
Original contributions Original contributions Boroday, N.V. Chekhun, V.F. Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells Experimental Oncology |
| description |
Background: The mechanisms of drug resistance of cancer have not been yet elucidated in details. Recently, the role of mast cells (MCs) in the development of drug resistance has been brought in the limelight. The aim of the study was to examine the morphological features of doxorubicin (DOX)-resistant Walker 256 carcinosarcoma and to assess the response of MCs and histamine content in these cells in relation to the development of resistance to DOX as well as in DOX-resistant tumors. Materials and Methods: The DOX resistance was induced by serial passages of Walker 256 carcinosarcoma in rats in the setting of DOX treatment in vivo. MCs in tumors were detected in the sections by staining with Toluidine Blue O. Histamine content in MCs stained with solution of Water Blue-Orcein was assessed by Astaldi semiquantitative method taking into account different staining intensity. Results: Formation of DOX resistance in the course of serial passages of Walker 256 carcinosarcoma was accompanied by the increase in the number of MCs in tumors and histamine content. Nevertheless, in tumors with phenotype of complete DOX resistance the number of histamine-containing MCs decreased to the same level as in tumors of the original strain that are DOX-sensitive. Conclusion: MCs are involved in formation of DOX resistance in Walker 256 carcinosarcoma. |
| format |
Article |
| author |
Boroday, N.V. Chekhun, V.F. |
| author_facet |
Boroday, N.V. Chekhun, V.F. |
| author_sort |
Boroday, N.V. |
| title |
Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells |
| title_short |
Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells |
| title_full |
Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells |
| title_fullStr |
Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells |
| title_full_unstemmed |
Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells |
| title_sort |
morphological features of doxorubicin-resistant walker 256 carcinosarcoma and response of mast cells |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2018 |
| topic_facet |
Original contributions |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/139246 |
| citation_txt |
Morphological features of doxorubicin-resistant Walker 256 carcinosarcoma and response of mast cells / N.V. Boroday, V.F. Chekhun // Experimental Oncology. — 2018 — Т. 40, № 1. — С. 42-47. — Бібліогр.: 30 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
AT borodaynv morphologicalfeaturesofdoxorubicinresistantwalker256carcinosarcomaandresponseofmastcells AT chekhunvf morphologicalfeaturesofdoxorubicinresistantwalker256carcinosarcomaandresponseofmastcells |
| first_indexed |
2025-07-10T07:52:53Z |
| last_indexed |
2025-07-10T07:52:53Z |
| _version_ |
1837245639746912256 |
| fulltext |
42 Experimental Oncology 40, 42–47, 2018 (March)
MORPHOLOGICAL FEATURES OF DOXORUBICIN-RESISTANT
WALKER 256 CARCINOSARCOMA
AND RESPONSE OF MAST CELLS
N.V. Boroday*, V.F. Chekhun
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine,
Kyiv 03022, Ukraine
Background: The mechanisms of drug resistance of cancer have not been yet elucidated in details. Recently, the role of mast cells
(MCs) in the development of drug resistance has been brought in the limelight. The aim of the study was to examine the morphologi-
cal features of doxorubicin (DOX)-resistant Walker 256 carcinosarcoma and to assess the response of MCs and histamine content
in these cells in relation to the development of resistance to DOX as well as in DOX-resistant tumors. Materials and Methods:
The DOX resistance was induced by serial passages of Walker 256 carcinosarcoma in rats in the setting of DOX treatment in vivo.
MCs in tumors were detected in the sections by staining with Toluidine Blue O. Histamine content in MCs stained with solution
of Water Blue-Orcein was assessed by Astaldi semiquantitative method taking into account different staining intensity. Results:
Formation of DOX resistance in the course of serial passages of Walker 256 carcinosarcoma was accompanied by the increase in the
number of MCs in tumors and histamine content. Nevertheless, in tumors with phenotype of complete DOX resistance the number
of histamine-containing MCs decreased to the same level as in tumors of the original strain that are DOX-sensitive. Conclusion:
MCs are involved in formation of DOX resistance in Walker 256 carcinosarcoma.
Key Words: mast cells, drug resistance, doxorubicin, histamine.
Cancer drug resistance is one of the main factors
limiting effectiveness of antineoplastic therapy. The
mechanisms of both primary resistance featuring
the intrinsic properties of cancer cells and acquired
resistance arising as an adaptive response to che-
motherapeutics of various groups are in the spotlight.
Nevertheless, the mechanisms involved in the devel-
opment of the resistance to anthracyclines have not
been studied in depth.
Besides, in recent years, interest in properties
of the mast cells (MCs) has increased due to their
multifunctionality and involvement in adaptive re-
sponses and pathological processes. MCs population
is heterogeneous. According to their protease content,
human MCs have been divided into two phenotypes:
those containing only tryptase, termed MCT, and
those containing both tryptase and chymase, termed
MCTC. MCT are found in mucosa while MCTC are con-
sidered as MCs of connective tissue type [1]. MCs are
heterogeneous by many other factors: localizations,
content of granules, response to different stimulation
and pharmacological agents [2]. Nevertheless, such
classification is rather conventional since MCs can
dynamically change their properties according to the
conditions of microenvironment [1]. On their surface,
MCs express receptors to chemokines, immunoglobu-
lins (IgA, IgE, IgG), adrenaline, adenosine, estrogen,
leptin, histamine, serotonin, stem cell factor, etc [2].
MCs are capable of affecting tumor development,
angiogenesis, and adaptive immune reactions [3, 4].
Due to proteases, MCs are involved in stroma remod-
elling, which promotes invasion and metastasizing
of tumor cells [5, 6]. It is shown that density of MCs
increases with tumor progression [7]. Nevertheless,
there is no consistent view on the mechanisms of how
MCs affect tumor growth. Two hypotheses of interac-
tions between tumor and MCs exist. According to the
first hypothesis, MCs stimulate carcinogenesis due
to the expression of proteases, angiogenic and growth
factors. According to the second hypothesis, MCs,
on the contrary, possess cancer suppressive proper-
ties. The biological effects of MCs are mediated by the
range of substances released by these cells. Therefore,
the net effect of MCs on tumor growth depends on the
complex interactions between these substances and
stroma cells (endotheliocytes and fibroblasts) [8]. The
heterogeneous phenotype both of cancer cells and
MCs as well as the opposite effects of various factors
in such interplay should also be taken into account.
Histamine is one of the mediators involved in regu-
lation of wide range of physiological and pathological
processes in which MCs are the main factors. This low-
molecular monoamine participates in cell proliferation
and differentiation, hematopoiesis, regeneration,
wound healing, signal transduction in aminergic neu-
rons, and also in a number of brain functions, in inflam-
mation and modulation of the immune response [9].
Histamine plays an important role in physiological
and pathological processes in mammary glands being
involved in growth regulation, differentiation and func-
tion in pregnancy and lactation [10]. Four subtypes
of histamine receptors (H1, H2, H3 and H4) have been
revealed in mammary glands. Activation of H2 and
H3 receptors promotes proliferation of cancer cells
in vitro, while activation of H1 and H4 receptors inhib-
its such proliferation [11]. Also, histamine may affect
tumor indirectly via activation of angiogenesis [12].
Histamine possesses both proangiogenic and antian-
Submitted: December 21, 2017
Correspondence: E-mail: boroday1@ukr.net
Abbreviations used: DOX — doxorubicin; MCs — mast cells.
Exp Oncol 2018
40, 1, 42–47
Experimental Oncology 40, 42–47, 2018 (March)40, 42–47, 2018 (March) (March) 43
giogenic properties depending on its concentration,
availability of cofactors and tumor microenvironment.
Nevertheless, histamine seems to affect angiogenesis
not directly but via endothelial cells in cooperation with
other proangiogenic factors.
The role of histamine in metastasizing is well known.
The expression of MMR-2 and MMR-9 in breast cancer
cells in vitro varies depending on histamine concentra-
tion, this effect being mediated by H2 and H4 [13]. Due
to H1 activation, histamine modulates MMR-2 activity
not only in cancer cells but also in fibroblasts [14].
The role of MCs in the development of resistance
to chemotherapeutic agents has not yet been clarified.
Study of the factors triggering doxorubicin (DOX) resis-
tance suggests that tumor cell-to-extracellular matrix
interactions are important. The purpose of the study
was to examine the morphological features of DOX-
resistant Walker 256 carcinosarcoma and to assess
the response of MCs and histamine content in these
cells in relation to the development of DOX resistance.
MATERIALS AND METHODS
The rats from the animal facility of R.E. Kavetsky
Institute of Experimental Pathology, Oncology and
Radiobio logy, the National Academy of Sciences
of Ukraine (Kyiv, Ukraine) were used in the study. The
use and care of the experimental animals was per-
formed in accordance with the standard international
rules of biologic ethics and was approved by Institu-
tional Animal Care and Use Committee. Wal ker 256 car-
cinosarcoma was obtained from National Repository
of Cell Lines and Transplanted Tumors of R.E. Kavetsky
Institute of Experimental Pathology, Oncology and
Radiobio logy. The rats were inoculated with original
Walker 256 tumor cells or cells of DOX- resistant strain.
Walker 256 carcinosarcoma with induced resistance
to DOX was obtained as described earlier by serial
transplantation of tumor cells in DOX-treated animals
(12 in vivo passages in total) [15, 16]. The tumors of the
parental strain, the tumors after four in vivo passages
in DOX setting (partial DOX resistance), and the tumors
of the refractory phenotype (complete DOX resistance)
were studied. The separate group of the rats bearing
Walker 256 carcinosarcoma of the refractory phe-
notype was treated with DOX at a dose of 1.5 mg/kg
(accumulated dose of 7.5 mg/kg), and such tumors
were also evaluated.
MCs in tumors were detected by staining with 1%
solution of Toluidine Blue O, (Sigma-Aldrich, USA)
in 0.5 M HCl by the standard technique [17] and
counted as per 1 mm2 of tumor tissue [18].
Histamine in MCs was detected in the sections
stained with solution of Water Blue — Orcein [19].
The results of cytochemical reaction were expressed
as a percentage of positive cells [20]. Histamine con-
tent in MCs was assessed by Astaldi semiquantitative
method, taking into account different staining intensity
(weak, moderate or intensive) in 100 cells.
The statistical analysis of data was carried out
by method of variation statistics using Microsoft Excel
2010 (Microsoft Corp., USA). The arithmetic mean
and its error (M ± m) were calculated. The statistical
significances of differences between mean values
were assessed with Student’s t-test. Differences
at p < 0.05 were considered significant.
RESULTS
Morphological features of Walker 256 carci-
nosarcoma. In 6 days after transplantation of paren-
tal strain of Walker 256 carcinosarcoma, tumor node
of 2.3 ± 0.3 cm was evident. Tumor cells were bunched
in bundles and layers or solid complexes plunged into
muscular tissue here and there and sprouted along
muscle fibers and blood vessels disarranging the struc-
tural architectonics of skeletal muscles. Sarcomatous
cells with hyperchromic nuclei varied in form (spindle,
stellated) and size. Epithelioid cells were distinguished
by moderate polymorphism and had mainly hypochro-
mic nuclei. In these cells, individual mitoses have been
observed. On the periphery of tumor node, epithelioid
cells formed so-called pseudo-follicular structures and
solid layers. The tumors were, as a rule, limited to the
surrounding sarcomatous stroma (Fig. 1). At the center
of the tumor, the tissue was less structured than on the
periphery, polymorphism of cells and nuclei was less
expressed. Necrotic foci of various sizes were observed.
Fig. 1. Walker 256 carcinosarcoma: pseudo-follicular structures (1)
and solid bundles (2) (stained with hematoxylin and eosin, x 200)
In 16–18 days after transplantation, the tumor
node took on more structured patterns with formation
of pseudo-follicular structures and outgrowth of the
bundles of sarcomatous cells with light epithelioid
cells located between them. At sites of proliferation,
active angiogenesis with ingrowth of blood vessels
into the tumor has been observed. The remaining
muscle fibers were lysed. In the center of tumor node,
necrotic foci have been observed. The increased
number of apoptotic cells has been evident.
MCs in Walker 256 carcinosarcoma tumors.
A small number of MCs was detected at the central
sites of the tumors (6.7 ± 1.3 cells per 1 mm2), some
of them with metachromatically stained granularity;
some grains sticking together (Fig. 2). Most of these
cells were destroyed partially or completely. In MCs
that seem to be intact, specific granules exited outside.
44 Experimental Oncology 40, 42–47, 2018 (March)
Fig. 2. MCs in a section of Walker 256 carcinosarcoma (control)
(stained with Toluidine Blue O, × 400)
It is necessary to notice that MC count (up to 15–
20 per 1 mm2) increased when the inflammation foci
were observed. In such cases, MCs were larger and
contained specific metachromatic granularity in a cy-
toplasm. Observed degranulation of MCs with the
release of the specific granules seems to indicate their
functional activity. Such cells were found mainly at the
peripheral sites of tumor node.
Morphological changes of Walker 256 carcino-
sarcoma throughout formation of DOX resistance
and in DOX-resistant tumors upon DOX treatment.
In tumors with partial DOX resistance (four in vivo
passages in DOX setting), tumor cells were less poly-
morphic. Light epithelial cells in clusters prevailed,
in particular, on the periphery of tumor node. At the
central sites of tumor, cells with moderate nuclear
polymorphism were predominant along with the ne-
crotic foci. The number of blood vessels of capillary
type increased. Also, in comparison with original
tumors, partially resistant tumors were characterized
by more structured patterns with formation of pseudo-
follicular structures and the cords of sarcomatous cells
with dark hyperchromatic nuclei.
In tumors with complete DOX resistance, vascular-
ization further enhanced, mainly at the expense of the
vessels of capillary type with increasing hemorrhages
accompanying with infiltration and plasmatic suffu-
sion of the tumor. In the central zones of the tumor,
the necrotic foci of various size, mainly infiltrated with
neutrophils and monocytes were observed. On the
periphery, tumor cells were bunched in bundles and
layers of solid complexes that plunged into muscular
tissue, while the remnants of muscles were also evident
(Fig. 3). Epithelioid cells of moderate polymorphism
and with prevalence of hypochromic nuclei prevailed
and formed pseudofollicular structures and solid lay-
ers with bundles of sarcomatous cells. The number
of mitoses in these cells increased. The pathological
mitoses were also evident.
DOX treatment of the rats bearing DOX-resistant
Walker 256 tumors has not changed substantially
the morphology of tumors. Epithelioid cells remained
prevalent but fibrotization increased in comparison
with resistant type (Fig. 4). Furthermore, the signifi-
cant vascularization mainly at the expense of capillary
vessels and increasing numbers of hemorrhages was
evident.
Fig. 3. The cords of Walker 256 carcinosarcoma DOX-resistant
tumor cells growing into muscular tissue (stained with hema-
toxylin and eosin, × 400)
Thus, with increasing DOX resistance of Walk-
er 256 carcinosarcoma the number of capillary vessels
increased, epithelioid cells being the main component
of tumors. The fibrotization of tumor node was also
observed. Upon DOX treatment of DOX-resistant type
of Walker 256 carcinosarcoma, the features above
became more prominent.
Fig. 4. Elements of connective tissue (1), hemorrhages (2),
vessels (arrows) in DOX-resistant Walker 256 carcinosarcoma
upon DOX treatment (stained with hematoxylin and eosin, × 400)
MCs in Walker 256 carcinosarcoma throughout
formation of DOX resistance and in DOX-resistant
tumors upon DOX treatment. In tumors with partial
DOX resistance, MCs located mainly in connective
tissue layers. Occasionally, MCs were observed near
blood vessels. Granules in cytoplasm did not stick
together suggesting MC metabolical activation. Some
granules were found extracellularly while MCs remain
intact.
In tumors with complete DOX resistance, MCs lo-
cated centrally or among pseudo-follicular structures,
or near blood vessels. The insignificant part of such
cells was degranulated with the granules exiting the
cells while a fraction of granules remaining in a cy-
Experimental Oncology 40, 42–47, 2018 (March)40, 42–47, 2018 (March) (March) 45
toplasm (Fig. 5). MCs were also found in connective
tissue structures surrounding tumor node.
Fig. 5. MCs (arrows) in central zones of DOX-resistant Wal-
ker 256 carcinosarcoma (stained with Toluidine Blue O, × 400)
After DOX treatment, the number of MCs in DOX-
resistant tumors increased compared to that in DOX-
resistant tumors of animals which were not DOX treated
(Fig. 6). These cells localized predominantly in the
connective tissue components of tumors and near the
vessels of capillary type crowded with erythrocytes.
In cytoplasm of the most part of these MCs, disperse
granules were visible with increased number of the
degranulated cells wherein specific granules being
released extracellularly.
0
5
10
15
20
Control
*
*
Partial DOX
resistance
Complete DOX
resistance
Complete DOX
resistance + DOX
treatment
M
C
nu
m
be
r p
er
1
m
m
2
Fig. 6. The number of MCs in DOX-resistant Walker 256 carcinosar-
coma in animals treated or non-treated with DOX. *р ≤ 0.05 com-
paring with control
Histamine content in MCs in Walker 256 car-
cinosarcoma throughout formation of DOX resis-
tance and in DOX-resistant tumors upon DOX treat-
ment. In animals bearing original Walker 256 tumor,
histamine-containing MCs were observed mainly near
blood vessels, on the periphery of tumor node, and also
at the sites where tumor cords grew into the muscles
(Fig. 7). In partially resistant tumors, the number of his-
tamine-containing MCs increased significantly (Fig. 8,
9) while the count of MCs in completely resistant tumors
dropped to the initial level in the original tumor (Fig. 9).
DOX treatment of animals with DOX-resistant tumor
did not further affect histamine-containing MC count.
The percentage of histamine-positive MCs with inten-
sive staining increased with increasing DOX resistance
with the concomitant decrease in the number of moder-
ate and poorly stained histamine-positive MCs (Fig. 10).
DISCUSSION
As was shown earlier, the formation of complete
DOX resistance of Walker 56 carcinosarcoma in rats
requires 12 passages in the setting of DOX treatment
in vivo [15]. Four courses of chemotherapy result
in partial DOX resistance with tumor growth inhibi-
tion by about 30%, while DOX treatment inhibited
the growth of parental Walker-256 carcinosarcoma
by about 65% [16]. These two time-points correspond-
ing to partial (4 passages) and complete (12 pas-
sages) DOX resistance were selected for studying the
morphological features of Walker 256 carcinosarcoma
Fig. 7. MCs (arrows) at the site of tumor ingrowth into muscular
tissue (stained with Water Blue — Orcein, × 200)
Fig. 8. MCs (arrows) in central zones of Walker 256 carcinosar-
coma with phenotype of partial resistance (stained with Water
Blue — Orcein, × 200)
Control Partial DOX
resistance
Complete DOX
resistance
Complete DOX
resistance + DOX
treatment
Nu
m
be
r o
f h
is
ta
m
in
e-
co
nt
ai
ni
ng
M
Cs
pe
r 1
m
m
2
0
5
10
15
20
25
30
*
Fig. 9. The number of histamine-containing MCs in DOX-
resistant Walker 256 carcinosarcoma in animals treated or non-
treated with DOX. *р ≤ 0.05 comparing with control
46 Experimental Oncology 40, 42–47, 2018 (March)
and the biology of MCs in dynamics of the formation
of DOX resistance.
Morphologically, in DOX-resistant tumors we have
found more structured patterns with formation of pseu-
do-follicular structures, slight outgrowth of the bundles
of sarcomatous cells, and the activation of neoan-
giogenesis. MCs response was also evident with the
increasing functional activity of these cells manifested
as their degranulation with the release of the specific
granules. The data obtained correspond to the re-
sults of other researchers who showed the increased
MC count, basically, at the peripheral sites of a tumor
and surrounding tissues [21, 22] with degranulation
of single MCs in the centre of tumors [23]. The slight
increase in the number of these cells with small content
of intracytoplasmic granules was observable mainly
near small vessels.
In DOX-resistant tumors, the outgrowth of connec-
tive tissue component with fibrotization of tumor node
was noted. This fact can be explained by MC contri-
bution to remodeling of connective tissue structures
due to the activity of the matrix proteinases and
proteinases located in MC granules as well as the
intensification of collagen synthesis by activated fi-
broblasts [24]. The significant vascularization in DOX-
resistant tumors seemed to be associated in part with
known MC effects on angiogenesis due to histamine
and VEGF [25, 26].
Two biologically active agents produced by MCs,
heparin and histamine, acting oppositely play an im-
portant role in regulation of MC effects [27]. The an-
tagonism of these substances is the cornerstone of the
functional duality of MCs acting both in stimulation and
inhibition mode.
The number of MCs in DOX-resistant tumors se-
quentially increased, and the number of MCs contain-
ing histamine decreased. It is known that DOX may
stimulate secretion of histamine by MCs [28, 29] simi-
larly to substance 48/80, which leads to degranulation
of MCs. This occurs because heparin and chondroitin
sulfate are protein glycanes with strong negative
charge while DOX has a positive charge facilitating its
binding with heparin and protein glycanes that, in turn,
stimulates histamine release from MCs.
The phenotype of DOX resistance in Walk-
er 256 carcinosarcoma is associated also with
changes in chemical signals produced by the
cells of resistant tumors. The mediators secreted
by tumor cells may affect immunocompetent cells
changing the patterns of their synthesis and se-
cretion [30]. In this setting, the response of MCs
is expressed not only as an increase in MC number
but also in changing patterns of mediators located
in cytoplasm of these cells (both preformed and
newly synthesized). In particular, we have shown
that histamine content in MCs upon DOX treatment
decreased. But the decrease in histamine content
does not imply the decrease in their functional ac-
tivity. On the contrary, histamine is released from
cytoplasmatic granules into extracellular space. The
released histamine may enhance proliferation of tu-
mor cells, stimulate angiogenesis, increase activity
of matrix metalloproteinases (ММР-2 and ММР-9)
and provide for immunomodulating effect on im-
mune competent cells (macrophages, T-cells) [9,
13]. As to the stimulation of angiogenesis, we in fact
demonstrated the increased density of capillaries
in DOX-resistant tumors. Also, histamine is capable
to increase expression of periostin and collagen
І in fibroblasts [31] possibly contributing in the in-
creased fibrotization of tumor node demonstrated
in DOX-resistant tumors.
Our data are in line with the current opinion that
MСs are important in regulating different functions
relevant to pathogenesis of tumors and adaptation
of tumor-bearing host to the extreme factors including
chemotherapeutical agents. We have shown that MCs
involved in extracellular matrix remodeling contribute
to the formation of DOX resistance of Walker 256 car-
cinosarcoma.
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ls
Control
Partial DOX resistance
Complete DOX resistance
Complete DOX resistance + DOX
*
*
* * *
*
*
*
Fig. 10. Distribution of histamine-containing cells by staining
intensity.*р ≤ 0.05 comparing with control
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