Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma
Significant variability of anticancer efficacy of dichloroacetate (DCA) stimulated an active search for the agents capable to enhance it antitumor action. Therefore, the aim of this work is the study of capability of aconitine-containing antiangiogenic agent BC1 to enhance anticancer activity of DCA...
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| Cite this: | Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma / O.N. Pyaskovskaya, I.V. Boychuk, A.G. Fedorchuk, D.L. Kolesnik, O.I. Dasyukevich, G.I. Solyanik // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 192-196. — Бібліогр.: 27 назв. — англ. |
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Pyaskovskaya, O.N. Boychuk, I.V. Fedorchuk, A.G. Kolesnik, D.L. Dasyukevich, O.I. Solyanik, G.I. 2019-01-22T12:57:25Z 2019-01-22T12:57:25Z 2015 Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma / O.N. Pyaskovskaya, I.V. Boychuk, A.G. Fedorchuk, D.L. Kolesnik, O.I. Dasyukevich, G.I. Solyanik // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 192-196. — Бібліогр.: 27 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/145487 Significant variability of anticancer efficacy of dichloroacetate (DCA) stimulated an active search for the agents capable to enhance it antitumor action. Therefore, the aim of this work is the study of capability of aconitine-containing antiangiogenic agent BC1 to enhance anticancer activity of DCA against Ehrlich carcinoma. Materials and Methods: DCA (total dose was 1.3 g/kg of b.w.) and BC1 (total dose was 0.9 mg/kg of b.w.) were administered per os starting from the 2nd and 3rd days, respectively (8 administrations for each agent). Antitumor efficacy of agents was estimated. Lactate level, LDH activity and the state of mitochondrial electron transport chain in tumor cells as well as phagocytic activity and reactive oxygen species (ROS) production of tumor-associated macrophages (TAM) were studied. Results: Combined administration of DCA and ВС1 resulted in 89.8% tumor growth inhibition (p < 0.001), what is by 22.5% (p < 0.05) higher that that of DCA alone. This combined treatment was accompanied with a decrease of lactate level in tumor tissue by 30% (p < 0.05) and significant elevation of LDH activity by 70% (p < 0.01). Increased level of NO-Fe-S clusters and 2-fold reduction of Fe-S cluster content were revealed in tumor tissue of mice after DCA and BC1 administration. It was shown that combined therapy did not effect TAM quantity and their phagocytic activity but stimulated ROS production by TAMs by 78% (p < 0.05) compared to this index in control animals. Conclusion: Antiangiogenic agent ВС1 in combination with DCA considerably enhances antitumor activity of DCA via significant decrease of Fe-S-containing protein level resulted from substantial elevation of nitrosylation of these proteins. Key Words: Ehrlich carcinoma, dichloroacetate, aconitine-containing agent, tumor-associated macrophages, mitochondrial electron transport chain. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma Article published earlier |
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Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma |
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Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma Pyaskovskaya, O.N. Boychuk, I.V. Fedorchuk, A.G. Kolesnik, D.L. Dasyukevich, O.I. Solyanik, G.I. Original contributions |
| title_short |
Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma |
| title_full |
Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma |
| title_fullStr |
Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma |
| title_full_unstemmed |
Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma |
| title_sort |
aconitine-containing agent enhances antitumor activity of dichloroacetate against ehrlich carcinoma |
| author |
Pyaskovskaya, O.N. Boychuk, I.V. Fedorchuk, A.G. Kolesnik, D.L. Dasyukevich, O.I. Solyanik, G.I. |
| author_facet |
Pyaskovskaya, O.N. Boychuk, I.V. Fedorchuk, A.G. Kolesnik, D.L. Dasyukevich, O.I. Solyanik, G.I. |
| topic |
Original contributions |
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Original contributions |
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2015 |
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Experimental Oncology |
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Article |
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Significant variability of anticancer efficacy of dichloroacetate (DCA) stimulated an active search for the agents capable to enhance it antitumor action. Therefore, the aim of this work is the study of capability of aconitine-containing antiangiogenic agent BC1 to enhance anticancer activity of DCA against Ehrlich carcinoma. Materials and Methods: DCA (total dose was 1.3 g/kg of b.w.) and BC1 (total dose was 0.9 mg/kg of b.w.) were administered per os starting from the 2nd and 3rd days, respectively (8 administrations for each agent). Antitumor efficacy of agents was estimated. Lactate level, LDH activity and the state of mitochondrial electron transport chain in tumor cells as well as phagocytic activity and reactive oxygen species (ROS) production of tumor-associated macrophages (TAM) were studied. Results: Combined administration of DCA and ВС1 resulted in 89.8% tumor growth inhibition (p < 0.001), what is by 22.5% (p < 0.05) higher that that of DCA alone. This combined treatment was accompanied with a decrease of lactate level in tumor tissue by 30% (p < 0.05) and significant elevation of LDH activity by 70% (p < 0.01). Increased level of NO-Fe-S clusters and 2-fold reduction of Fe-S cluster content were revealed in tumor tissue of mice after DCA and BC1 administration. It was shown that combined therapy did not effect TAM quantity and their phagocytic activity but stimulated ROS production by TAMs by 78% (p < 0.05) compared to this index in control animals. Conclusion: Antiangiogenic agent ВС1 in combination with DCA considerably enhances antitumor activity of DCA via significant decrease of Fe-S-containing protein level resulted from substantial elevation of nitrosylation of these proteins. Key Words: Ehrlich carcinoma, dichloroacetate, aconitine-containing agent, tumor-associated macrophages, mitochondrial electron transport chain.
|
| issn |
1812-9269 |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/145487 |
| citation_txt |
Aconitine-containing agent enhances antitumor activity of dichloroacetate against Ehrlich carcinoma / O.N. Pyaskovskaya, I.V. Boychuk, A.G. Fedorchuk, D.L. Kolesnik, O.I. Dasyukevich, G.I. Solyanik // Experimental Oncology. — 2015. — Т. 37, № 3. — С. 192-196. — Бібліогр.: 27 назв. — англ. |
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| fulltext |
192 Experimental Oncology 37, 192–196, 2015 (September)
ACONITINE-CONTAINING AGENT ENHANCES ANTITUMOR
ACTIVITY OF DICHLOROACETATE AGAINST EHRLICH CARCINOMA
O.N. Pyaskovskaya, I.V. Boychuk, A.G. Fedorchuk, D.L. Kolesnik, O.I. Dasyukevich, G.I. Solyanik*
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv
03022, Ukraine
Significant variability of anticancer efficacy of dichloroacetate (DCA) stimulated an active search for the agents capable to enhance
it antitumor action. Therefore, the aim of this work is the study of capability of aconitine-containing antiangiogenic agent BC1 to en-
hance anticancer activity of DCA against Ehrlich carcinoma. Materials and Methods: DCA (total dose was 1.3 g/kg of b.w.) and
BC1 (total dose was 0.9 mg/kg of b.w.) were administered per os starting from the 2nd and 3rd days, respectively (8 admini strations
for each agent). Antitumor efficacy of agents was estimated. Lactate level, LDH activity and the state of mitochondrial electron
transport chain in tumor cells as well as phagocytic activity and reactive oxygen species (ROS) production of tumor-associated
macrophages (TAM) were studied. Results: Combined administration of DCA and ВС1 resulted in 89.8% tumor growth inhibition
(p < 0.001), what is by 22.5% (p < 0.05) higher that that of DCA alone. This combined treatment was accompanied with a decrease
of lactate level in tumor tissue by 30% (p < 0.05) and significant elevation of LDH activity by 70% (p < 0.01). Increased level
of NO-Fe-S clusters and 2-fold reduction of Fe-S cluster content were revealed in tumor tissue of mice after DCA and BC1 ad-
ministration. It was shown that combined therapy did not effect TAM quantity and their phagocytic activity but stimulated ROS
production by TAMs by 78% (p < 0.05) compared to this index in control animals. Conclusion: Antiangiogenic agent ВС1 in com-
bination with DCA considerably enhances antitumor activity of DCA via significant decrease of Fe-S-containing protein level re-
sulted from substantial elevation of nitrosylation of these proteins.
Key Words: Ehrlich carcinoma, dichloroacetate, aconitine-containing agent, tumor-associated macrophages, mitochondrial
electron transport chain.
The achievements of recent years in regard
to genetic mechanisms of cell metabolic regulation,
mitochondrial functioning and synthesis of macromo-
lecules have turn an attention to energy metabolism
of tumor cells and Warburg effect [1]. Numerous
metabolic pathways which are supposed to be impor-
tant for tumor growth and progression are considered
as perspective targets for the development of effective
anticancer drugs [2, 3]. Among such drugs dichloro-
acetate (DCA) could be mentioned [4, 5]. It is known
that DCA inhibits pyruvate dehydrogenase kinase what
results in elevation of pyruvate dehydrogenase acti-
vity what in turn promotes the shift of cell metabolism
from glycolysis toward oxidative phosphorylation, i.e.
the state of tumor cells when their proliferative acti-
vity is decreased and apoptosis risk is enhanced [6,
7]. Apart from glycolysis inhibition, DCA may cause
significant decrease of lactacidosis level in tumor cell
microenvironment which is known to promote tumor
cell viability [8].
For 30 years DCA has been used for correction
of lacticemia, caused by high intensity of glycolysis
or defective cell respiration. In 2008 Canadian scien-
tists have shown that DCA is highly effective against
many tumor types and possesses low toxicity toward
normal tissues [9]. Such features of DCA have moti-
vated an initiation of active scientific studies of this
drug in oncology.
At present time, the data on anticancer efficacy
of DCA are in large part contradictory. From one
side, there are conclusive evidences obtained in ex-
periments in vitro and in vivo on feasibility of DCA use
as anticancer drug [10]. There has been documented
an ability of DCA to suppress proliferation of many
tumor cell lines with no impact on their death rates [6,
11]. Also, it has been shown that DCA induces apop-
tosis only at high doses and is active against cells with
defective electron-transport chain [12, 13]. From other
side, as it has been shown in preclinical studies DCA
is ineffective against many experimental tumor models
including lung cancer, neuroblastoma, lymphoma, sar-
coma [14]. An analysis of the results of clinical studies
performed in recent years has also revealed significant
variability of anticancer efficacy of DCA: from its high
efficacy to complete inefficacy [15, 16].
In the last years there is an active search for
the agents capable to enhance anticancer action
of DCA. Among such agents a special place is oc-
cupied by inhibitors of tumor angiogenesis providing
the deficiency of the main energetic substrates (glu-
cose and oxygen) via suppression of tumor vessel
development [17].
Therefore, the aim of this work is the study of ca-
pability of antiangiogenic agent to enhance anticancer
activity of DCA against Ehrlich carcinoma.
Aconitine-containing agent ВС1 has been used
as an inhibitor of tumor angiogenesis. As it has been
shown earlier, ВС1 exerts its antiangiogenic action via
inhibition of endothelial cell proliferation which under-
lies its significant antitumor activity against tumors with
angiogenesis-dependent growth [18, 19].
Submitted: May 18, 2015.
*Correspondence: E-mail: gsolyanik@gmail.com
Abbreviations used: DCA — dichloroacetate; LDH — lactate dehydro-
genase; MtETC — mitochondrial electron-transport chain; ROS — re-
active oxigen species; TAM — tumor-associated macrophages.
Exp Oncol 2015
37, 3, 192–196
Experimental Oncology 37, 192–196, 2015 (September) 193
MATERIALS AND METHODS
Experimental animals and tumor strain. The study
was carried out using 2.0–2.5 months white outbred
mice weighting 18.5–21 g, bred at animal facility
of R.E. Kavetsky Institute of Experimental Pathology,
Oncology and Radiobiology of the NAS of Ukraine
(Kyiv, Ukraine). Animal study protocols and opera-
tion procedures were carried out in accordance with
the main requirements to keeping and working with
laboratory animals and to the rules of local Bioethics
Committee.
E h r l i c h c a rc i n o m a c e l l s ( 1 . 2 • 10 6 c e l l s
in 0.2 ml of physiological solution per animal) were
transplanted subcutaneously in dorsal forelimb region.
Design of the study. After tumor cell transplanta-
tion the animals were distributed into 4 groups: control
one and three experimental (Table 1). 0.4 ml of aque-
ous DCA (Sigma, USA) solution were administered
per os to animals from 1st and 3rd groups in a day
starting from 2nd day after tumor cell transplantation
(8 administrations in total). The total DCA dose was
1.3 g/kg of animal body weight. The mice from 3rd
group were also treated with aconitine-containing
agent ВС1 (Aksomed, Ukraine) each second day
starting from 3rd day after tumor cell transplantation
(8 administrations in total). Total ВС1 dose (by aconi-
tine content) was 0.9 mg/kg of animal body weight.
The animals from 2nd group were administered with
ВС1 only, using the same dosage and regimen
of administration as these for 3rd group. Animals from
control group received water for injections by the cor-
responding scheme.
Table 1. The design of experiment
Groups of mice Number of mice/group Type of treatment
Control 12 H2O
Experimental-1 10 DCA
Experimental-2 7 ВС1
Experimental-3 10 DCA + ВС1
After tumor appearance its diameter has been
measured triply per week using caliper. Tumor volumes
were calculated by formula:
V = 3.14 • d3/8,
where d — tumor diameter.
Antitumor activity was evaluated at 17th day after
tumor cell transplantation with the use of tumor inhibi-
tion index (D), calculated by the formula:
D = (Vk – V) • 100% / Vk,
where Vk and V — average tumor volumes in control
and experimental groups, respectively.
At the 2nd day after termination of the treatment
(17 day of tumor growth) the animals were sacrificed
under light ether narcosis. Tumor tissue was isolated.
Then the effect of DCA on lactate level, lactate dehy-
drogenase (LDH) activity, functional state of mitochon-
drial electron-transport chain (MtETC), counts and
functional activity of tumor-associated macrophages
(TAM) have been analyzed. Each index has been evalu-
ated in tumor tissue from 4–6 mice per each group.
Lactate level in tumor tissue was determined
by spectrophotometry using LDH (Sigma, USA). The
method is based on the reaction of lactate oxidation
into pyruvate by LDH resulting in generation of NADH
from NAD [20].
Total LDH activity in frozen tumor tissue was mea-
sured by spectrophotometric method which is based
on determination of enzyme kinetics when pyruvate
is converted to lactate [20].
For analysis of functional activity of MtETC
in tumor cells, tumor tissue was cut into the samples
of cylindrical shape (d = 4.0 mm, l = 25–35 mm), frozen
and stored at −70 °C. EPR analysis of the samples was
performed at 77 К using spectrometer Е-109 Varian
(USA) at potential sweep speed of 500 Е/min, modu-
lation amplitude of 1.25•10 Е, power of super-high-
frequency radiation of 10.0 mW, constant session
of apparatus of 1.0 s.
The levels of reduced non heme iron-sulfur (Fe-S)
centers (g = 1.94) of MtETC proteins, nitrosyl (NO)
complexes of heme iron (gсер = 2.01) and Fe-S nitrosyl
complexes (gсер = 2.03) were determined by the data
of EPR spectra.
Phagocytic activity (engulfing capacity) of TAM
was determined by flow cytometer FACSCalibur (Bec-
ton Dickinson, USA). TAMs were identified with the use
of anti-СD14 antibodies according to the instructions
of the manufacturer (Becton Dickinson, USA). Analysis
of phagocytic activity of СD14+ cells was performed
with the use of FITC-labeled staphylococcus [21].
Reactive oxigen species (ROS) production by TAM
was carried out using 2.7-dichlorodihydrofluorescein
diacetate (Sigma, USA) [21] by flow cytometry.
Statistical analysis of the data was performed
with the use of descriptive statistics, parametric
Student’s t-test and nonparametric Mann — Whitney
U test, with the use of Microsoft Excel, Microcal Origin
and Statistica. The data in the tables and figures are
presented as mean ± standard error.
RESULTS AND DISCUSSION
The performed studies have shown sufficiently high
antitumor activity of DCA against Ehrlich carcinoma.
The percent of tumor growth inhibition after the therapy
(17th day of tumor growth) achieved 73.3% (Table 2),
what is in complete accordance with the results pre-
sented in the study [22].
Table 2. Antitumor efficacy of DCA, BC1 and their combination against
Ehrlich carcinoma
Groups of mice Tumor volume, mm3 Inhibition index, %
Control 7828.0 ± 893.0 −
Experimental-1 2092.0 ± 380.3* 73.3
Experimental-2 5306.8 ± 1595.3 −
Experimental-3 805.1 ± 137.8* 89.8
Note: *p < 0.05.
In the group of animals treated with ВС1, there was
observed a tendency to inhibition of Ehrlich carcinoma
growth which did not reach statistical significance due
to variability of tumor volumes. This result is in agree-
ment with our previous data on anticancer activity
of BC1 against solid form of Ehrlich carcinoma. In this
study growth kinetic analysis has shown that adminis-
tration of BC1 (at the total dose of 0.9 mg/kg of b.w.)
194 Experimental Oncology 37, 192–196, 2015 (September)
did not affect tumor volume till 15th day and only after
that tumor inhibition indices constantly increased
growing up to 77.3% in a week after therapy comple-
tion [23].
Despite the fact that administration of antiangio-
genic aconitine-containing drug ВС1 to mice did not
cause significant inhibition of tumor growth, intro-
duction of this agent into DCA-based therapy signifi-
cantly enhanced anticancer action of DCA. Combined
administration of DCA and ВС1 resulted in 89.8%
tumor growth inhibition (p < 0.001), what is by 22.5%
(p < 0.05) higher that that of DCA alone (see Table 2).
Antitumor activity of DCA administered alone
or in combination with BC1 was accompanied with sta-
tistically significant decrease of lactate level in tumor tis-
sue by 22% (p < 0.05) and 30% (p < 0.05), respectively
(Fig. 1, a), which may be caused by activation of DCA-
dependent oxidative decarboxylation of pyruvate and
its conversion to acetyl-coenzyme А (the main substrate
for Krebs cycle). Adaptive reaction of the cell on lactate
level decrease is elevation of LDH activity; LDH activity
increase by 120% (p < 0.001) and 70% (p < 0.01) was
observed in mice treated with DCA and DCA + ВС1,
respectively (Fig. 1, b). Such activation of the enzyme
could not compensate completely the decrease of lac-
tate level at the background of DCA administration what
could be possibly caused by higher rate of the pyruvate
conversion to acetyl-coenzyme А than to lactate. In the
work [24] it has been shown that the DCA-dependent
decrease of lactate level and increase of LDH activity
in human colorectal cancer cells and prostate carci-
noma cells has been accompanied with significant
decrease of pyruvate-to-lactate exchange rate.
0
20
40
60
80
100
120
Control DCA DCA + BC1
Tu
m
or
la
ct
at
e
le
ve
l (
%
)
0
50
100
150
200
250
Control DCA DCA + BC1
LD
H
ac
tiv
ity
(%
)
*
*
*
*
Fig. 1. Lactate level (a) and LDH activity (b) in tumor tissue
of LLC/R9 bearing mice: control, DCA-treated and DCA + BC1-
treated groups. *p < 0.05 compared to control
Our study has shown that administration of DCA
alone or DCA in combination with ВС1 resulted in signifi-
cant increase of ROS production in tumor cells as well
as normal cells involved in tumor process (TAM). High
ROS production in tumor cells was manifested in en-
hancement of nitrosylation both heme proteins (with
generation of NO-heme complexes) and non-heme
proteins (with generation of dinitrosyl-iron complexes;
DNIC). The latest play a key role in MtETC functioning.
As one may see on Fig. 2, administration of DCA
alone or in combination with ВС1 caused statistically
significant elevation of DNIC level (Fig. 2, a) by 140%
(p < 0.05) and 180% (p < 0.01), as well as NO-heme
complexes (Fig. 2, b) by 85% (p < 0.05) and 43%
(p < 0.05), respectively.
0
50
100
150
200
250
300
350
Control DCA DCA + BC1
DN
IC
le
ve
l (
%
)
0
50
100
150
200
250
300
350
Control DCA DCA + BC1
Le
ve
l o
f N
O
-h
em
e
(%
)*
*
*
*
Fig. 2. Levels of DNIC (a) and NO-heme (b) in tumor tissue
of LLC/R9 bearing mice: control, DCA-treated and DCA + BC1-
treated groups. *p < 0.05 compared to control
It is known that generation of NO-Fe-S clusters
(DNIC) could lead to inactivation of Fe-S cluster
proteins in MtETC, disturb coupling of oxidative
phosphorylation with respiration and cause tumor
cell death [25]. As it is shown on Fig. 3, significant in-
crease of DNIC level in tumor tissue of mice after DCA
administration did not lead to decrease of Fe-S cluster
content, but at the same time Fe-S cluster content was
2-fold lower (p < 0.01) in tumor tissue of mice treated
with DCA and ВС1 than that indices in control group
of animals. These results have indicated that stimula-
tion of ROS production induced by DCA administration
did not cause MtETC dysfunction and had insignificant
impact in anticancer activity of this agent. In contrary,
stimulation of ROS production observed after com-
bined administration of DCA and ВС1 and decreased
levels of Fe-S clusters made a contribution in the
antitumor activity of combined therapy.
0
20
40
60
80
100
120
140
Control DCA DCA + BC1
Le
ve
l o
f F
e-
S
cl
us
te
rs
(%
)
*
Fig. 3. Levels of Fe-S clusters in tumor tissue of LLC/R9 be aring
mice: control, DCA-treated and DCA + BC1-treated groups.
*p < 0.01 compared to control
It is known that nitric oxide species appear in tumor
tissue via its increased synthesis by mitochondrial NO-
synthase (a constitutive isoform localized on internal
mitochondrial membrane) and also by inducible NO-
synthase of TAM.
TAMs develop from circulating monocytes recruited
in the region of tumor growth because recognition and
elimination of transformed cells is among physiologic
functions of immune system [26]. Recruited mono-
Experimental Oncology 37, 192–196, 2015 (September) 195
cytes in tumor lesion are capable to undergo functional
diversification in two populations: macrophages with
М2-phenotype which reveal proangiogenic action
promoting tumor growth and progression; macro-
phages with М1-phenotype which participate in acti-
vation of antitumor immunity and reveal an expressed
cytotoxic action against tumor cells [27]. Antitumor
action of TAMs could be exerted by two mechanisms:
phagocytic activity and ROS production.
Analysis of functional activity of TAM has shown
significant increase of ROS production level by 241.0%
(p < 0.01) in tumor tissue of mice after DCA admini-
stration at monoregimen compared to control ani-
mals (Fig. 4, c). The high level of ROS is determined
by an expressed tendency to increased TAM counts
in tumor tissue (Fig. 4, a) as well as activation of ROS
production by TAM. DCA administration also led
to activation of TAM phagocytic activity (Fig. 4, b):
in DCA-treated animals phagocytic activity of TAM
was by 70.5% (p < 0.05) higher than that in control
mice. Such activation of cytotoxic activity of TAM
evidences on significant impact of TAM on antitumor
activity of DCA.
0
50
100
150
200
250
300
Nu
m
be
r o
f T
AM
(%
)
0
50
100
150
200
250
En
gu
lfi
ng
c
ap
ac
ity
o
f T
AM
(%
)
0
100
200
300
400
500
Le
ve
l o
f R
O
S
(%
)
Co
nt
ro
l
DC
A
DC
A
+
BC
1
Co
nt
ro
l
DC
A
DC
A
+
BC
1
Co
nt
ro
l
DC
A
DC
A
+
BC
1
*
*
*
Fig. 4. Number of TAM (a), their phagocytic activity (b) and ROS
production (c): control, DCA-treated and DCA + BC1-treated
groups. *p < 0.05 compared to control
It is necessary to note that an effect of DCA com-
bined with ВС1 toward counts and functional activity
of TAM differed from that of DCA alone. As it is shown
on Fig. 4, the combined therapy has no effect on TAM
quantity and their phagocytic activity. After the com-
bined therapy ROS production by TAM increased
by 78% (p < 0.05) compared to this index in control
animals but was significantly lower compared to cor-
responding index in mice treated with DCA alone. So,
this fact points on insignificant impact of cytotoxic
activity of TAM in sufficiently high antitumor activity
of DCA in combination with ВС1.
In conclusion, the presented results have demon-
strated that DCA exerts significant antitumor activity
against Ehrlich carcinoma mainly via induced activa-
tion of cytotoxic activity of TAM. Antiangiogenic agent
ВС1 in combination with DCA significantly enhances
antitumor activity of DCA via significant decrease of Fe-
S-containing protein level resulted from significant
elevation of nitrosylation of these proteins.
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