Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis
Aim:To study the influence of polyamine metabolism inhibitors on the growth, metastasis and ornithine decarboxylase (ODC) activity of Lewis lung carcinoma. Materials and Methods: Experiments were performed on female mice C57Bl/6 with Lewis lung carcinoma. Nω-hydroxy-nor-arginine (nor-NOHA) and α-dif...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
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| Cite this: | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis / О.А. Samoilenko, O.A. Milinevska, O.V. Karnaushenko, V.A. Shlyakhovenko, S.P. Zaletok // Experimental Oncology. — 2015. — Т. 37, № 2. — С. 151-153. — Бібліогр.: 20 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1859612832427933696 |
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| author | Samoilenko, О.А. Milinevska, O.A. Karnaushenko, O.V. Shlyakhovenko, V.A. Zaletok, S.P. |
| author_facet | Samoilenko, О.А. Milinevska, O.A. Karnaushenko, O.V. Shlyakhovenko, V.A. Zaletok, S.P. |
| citation_txt | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis / О.А. Samoilenko, O.A. Milinevska, O.V. Karnaushenko, V.A. Shlyakhovenko, S.P. Zaletok // Experimental Oncology. — 2015. — Т. 37, № 2. — С. 151-153. — Бібліогр.: 20 назв. — англ. |
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| description | Aim:To study the influence of polyamine metabolism inhibitors on the growth, metastasis and ornithine decarboxylase (ODC) activity of Lewis lung carcinoma. Materials and Methods: Experiments were performed on female mice C57Bl/6 with Lewis lung carcinoma. Nω-hydroxy-nor-arginine (nor-NOHA) and α-difluoromethylornithine (DFMO) were used as arginase and ODC inhibitors, correspondently. Inhibition of tumor growth was calculated by comparison of tumor volume in the treated and control groups. The average number of metastases per animal in the group and the average volume of pulmonary metastases per animal in the group have been determined. Determination of ODC — the key enzyme of the polyamine synthesis — in the samples of experimental tumors was performed by method of Luqman S. Results: Administration of DFMO or it’s combination with nor-NOHA resulted in the decrease of tumor growth rate, number and volume of lung metastases and was accompanied with reduced ODC activity in tumor tissue. Сonclusion: Modifiers of polyamine metabolism may be considered as promising targeted cancer therapy. Key Words: Lewis lung carcinoma, metastases, polyamine metabolism, ornithine decarboxylase, α-difluoromethylornithine, Nωhydroxy-nor-arginine.
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Experimental Oncology 37, 151–153, 2015 (June) 151
EFFECT OF POLYAMINE METABOLISM INHIBITORS ON LEWIS
LUNG CARCINOMA GROWTH AND METASTASIS
О.А. Samoilenko*, O.A. Milinevska, O.V. Karnaushenko, V.A. Shlyakhovenko, S.P. Zaletok
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv
03022, Ukraine
Aim: To study the influence of polyamine metabolism inhibitors on the growth, metastasis and ornithine decarboxylase (ODC)
activity of Lewis lung carcinoma. Materials and Methods: Experiments were performed on female mice C57Bl/6 with Lewis lung
carcinoma. Nω-hydroxy-nor-arginine (nor-NOHA) and α-difluoromethylornithine (DFMO) were used as arginase and ODC
inhibitors, correspondently. Inhibition of tumor growth was calculated by comparison of tumor volume in the treated and control
groups. The average number of metastases per animal in the group and the average volume of pulmonary metastases per animal
in the group have been determined. Determination of ODC — the key enzyme of the polyamine synthesis — in the samples of ex-
perimental tumors was performed by method of Luqman S. Results: Administration of DFMO or it’s combination with nor-NOHA
resulted in the decrease of tumor growth rate, number and volume of lung metastases and was accompanied with reduced ODC
activity in tumor tissue. Сonclusion: Modifiers of polyamine metabolism may be considered as promising targeted cancer therapy.
Key Words: Lewis lung carcinoma, metastases, polyamine metabolism, ornithine decarboxylase, α-difluoromethylornithine, Nω-
hydroxy-nor-arginine.
Polyamines (PA) — putrescine, spermidine and
spermine — natural polycations, which play a crucial
role in many fundamental biological processes: DNA
replication, transcription, translation, membrane protein
function, protein folding and others [1–6]. The level
of PA largely vary during physiological and pathological
processes (embryogenesis, ageing [7–9], regenera-
tion, infections, malignant transformation [10, 11]).
The increased level of PA in tumor growth is used
as a marker for monitoring in course of a disease
and efficiency of therapy [12–14]. Inhibition the level
of PA by blocking the activity of the key enzyme for pu-
trescine synthesis ornithine decarboxylase (ODC) —
one of the most promising directions in the develop-
ment of new approaches to the cancer treatment.
PA metabolism is regulated at different levels by a com-
plex system of enzymes, which are prima rily enzymes
of PA synthesis and transformation. The modification
of genes regulating PA metabolism by external factors,
not studied enough. The aim of this work was to study
the effect of inhibitors of PA metabolism on the growth
and metastasis of experimental Lewis lung carcinoma
in mice.
MATERIALS AND METHODS
Experiments were per formed on female
C57Bl/6 mice 16.2–23 g. Tumor strain Lewis lung car-
cinoma was obtained from National Bank of Cell Lines
and Transplanted Tumors of R.E. Kavetsky Institute
of Experimental Pathology, Oncology and Radiobio-
logy, NAS of Ukraine (Kyiv, Ukraine). All experiments
were conducted in accordance to international rules
of work with laboratory animals [15]. Lewis lung car-
cinoma transplanted intramuscularly into the shin
of the right rear leg 3•105 tumor cells in a volume
of 0.2 ml of sterile isotonic sodium chloride solution.
Animals were kept on a normal diet with free access
to water. Each treatment group had 3–6 animals, con-
trol group — 4 mice.
Inhibitors applied: arginase inhibitor — Nω-
hydroxy-nor-arginine (nor-NOHA) (Cayman Chemical,
USA), inhibitor of ODC — α-difluoromethylornithine —
DFMO (Centre de Recherche Merrell International,
16 rue d’Ankara, 67084 Strasbourg Cedex, France).
nor-NOHA prepared immediately before use and
injected into the abdominal cavity of mice in a dose
of 60 mg/kg (1.2 mg per mouse in 0.2 ml of isotonic so-
dium chloride solution) starting 7 days after tumor trans-
plantation. A comparative analysis was performed versus
a group of non-treated tumor-bearing mice (control).
In each treatment group, 5 injections of an inhibitor
were made. Control animals injected with 0.2 ml of ste-
rile isotonic sodium chloride solution. The volume
of tumors registered throughout the experiment.
The animals were decapitated 1 day after the last in-
jection of the inhibitors that is 14th day, and on the 26th
day after tumor transplantation.
The kinetics of tumor growth evaluated by means
of measuring tumor diameters. Inhibition of tumor
growth was calculated by comparison of tumor vo lume
in the treated and control groups. To assess the inten-
sity of the process of metastasis was used the following
criteria: the average number of pulmonary metasta-
ses per animal in the group and the average volume
of metastases per animal in the group. In these studies,
the number and volume of lung metastases in un-
treated animals and in animals treated as described
above for the tumor growth delay studies were scored
on day 26 post-tumor implant. The number and volume
of metastases were counted in the transmitted light
Submitted: January 23, 2015.
*Correspondence: E-mail: a-samoilenko@ukr.net
Abbreviations used: DFMO — alpha-difluoromethylornithine; nor-
NOHA — Nω-hydroxy-nor-arginine; ODC — ornithine decarboxy-
lase; PA — polyamines.
Exp Oncol 2015
37, 2, 151–153
SHORT COMMUNICATIONS
152 Experimental Oncology 37, 151–153, 2015 (June)
after removal of the lung, fixing it between the glass
plates. Tumor volume was calculated by the formula:
V = a2 · b · π/6,
where a — short, b — longer diameter of tumor.
The metastas is inhibition index (MII) was calculated
by the formula:
MII = ((Ac − Ae)/Ac) · 100,
where Ac — the number of metastases to the lung
in mice of the control group and Ae — the frequency
of metastases in a treated group.
Determination of the ODC activity in the samples
of expe rimental tumors was performed by meth-
od S. Luqman [16, 17]. The method is based on the fact
that the correspon ding enzyme converts L-ornithine
hydrochloride (substrate) to the yellow colored pu-
trescine adduct soluble in pentanol, which measured
by spectrophotometer.
The statistic processing of obtained results was
conducted with the help of Student’s t-criterion. The
data were reported as the M ± m.
RESULTS AND DISCUSSION
Arginine is a precursor for ornithine which may
be then converted into putrescine — the first partici-
pant in metabolic transformations of PA [18]. It was
found that DFMO — specific inhibitor of ODC — pre-
vents progression of cancer cells in model systems
[19]. L-arginine is a nonessential amino acid that plays
a central role in several biological systems includ-
ing the immune response. L-arginine is metabolized
by arginase I and arginase II with farther creation of or-
nithine [20]. In our experiments, the effect of PA syn-
thesis inhibitors — DFMO and a competitive inhibitor
of ornithine synthesis nor-NOHA — on the Lewis lung
carcinoma growth has been performed. The results
are presented in Fig. 1, Table 1.
As can be seen, there is a significant inhibition of tu-
mor growth begining of the eleventh day of the experi-
ment. During the experiment, both inhibitors studied
demonstrate inhibi ting activity (within 28–43%).
In this regard, it was important to follow the changes
in the dynamics of tumor growth that presented in Fig. 1.
0
1000
2000
3000
4000
5000
6000
11 13 15 18 20 22 24 26
Day after transplantation
Tu
m
or
v
ol
um
e,
V
m
m
3
Control
Nor-NOHA
DFMO
DFMO+nor-NOHA
*
*
*
*
Fig. 1. Lewis lung carcinoma gro wth in mice under the influence
of inhibitors of PA metabolism. *Significant difference (p < 0.05)
with each experimental group (excluding the DFMO + nor-NOHA
group at the 11th day, see Table 1)
Assessing the growth rate of the primary tumor
node, it was found that average daily increase of tumor
volume in the control group of animals within 9 days
(11–20 days) was 497.2 ± 56.0 mm3. The use of both
studied inhibitors and their combination led to a sig-
nificant reduction in the rate of growth, t he latter was
about 300 mm3 d uring the same time period. Signifi-
cant difference in the impact on growth inhibition when
using different inhibitors was not found (Fig. 2).
Table 1. Inhibition of growth of Lewis lung carcinoma in mice under influ-
ence of inhibitors of PA metabolism
Groups
Nu
m
be
r
of
a
ni
m
al
s The volume of tumors, mm3 Inhibition, %
11th day 13th day 20th day 11th
day
13th
day
20th
day
Control 4 1067 ± 133 2123 ± 165 5542 ± 439 − − −
nor-NOHA 5 690 ± 129* 1194 ± 163* 3163 ± 662* 35.3 43.0 43.0
DFMO 4 633 ± 114* 1534 ± 232* 3305 ± 461* 40.7 28.0 40.0
D F M O +
nor-NOHA 3 754 ± 138 1439 ± 149* 3504 ± 281* 0 32.0 37.0
Note: *p < 0.05 as compared to control.
0
100
200
300
400
500
600
Control nor-NOHA DFMO DFMO + nor-NOHA
Tu
m
or
g
ro
wt
h
ra
te
, m
m
3 /d
ay
Fig. 2. T he rate of tumor growth of Lewis lung carcinoma in mice
during 11–20 days compared to control
It is known that transplanted Lewis lung carcinoma
refers to metastatic tumors, so anti-metastatic effect,
as to this tumor strain, is an important indicator of antitu-
mor effect. The results of the next section of experiments
devoted to examining the influence of inhibitors of PA me-
tabolism on the growth of lung metastases (Table 2).
Table 2. E ffect of inhibitors of PA metabolism on the number and volume
of metastases of Lewis lung carcinoma in mice (26th day)
Groups
Number
of ani-
mals
The average number
of metastases
per 1 animal
The average volume
of metastases
per 1 animal
n MІІ, % mm3 МІІ, %
Control 4 11.0 ± 2.2 − 160.1 ± 44.1 −
nor-NOHA 5 16.6 ± 3.2 0 138.8 ± 44.2 0
DFMO 4 12.0 ± 0.7 0 9.8 ± 2.3* 94
DFMO + nor-NOHA 3 11.0 ± 2.3 0 11.9 ± 5.3* 93
Note: *p < 0.01 as compared to control.
As it is seen, the number of metastases in the
lungs of all treated mice did not differ versus control.
At the same time, volume of the metastatic lesions
in the DFMO and DFMO + nor-NOHA groups was near
90% less versus control.
Taking in mind the importance of the ODC enzyme
and PA synthesis for cell proliferation, the next sec-
tion of research was devoted to study of ODC activity
in the cells of Lewis lung cancer after the action of in-
hibitors of PA metabolism. The experimental results
are presented in Table 3.
The data obtained show that ODC activity in the tumor
increases during the tumor development. On 26th day
of experiment, ODC activity in tumors in all treated groups
Experimental Oncology 37, 151–153, 2015 (June) 153
was decreased versus control (see Table 3). It may
be expected that inhibition of ODC activity is the main
cause of inhibition of tumor growth. Our preclinical data
indicate that DFMO in combination with inhibitors of ar-
ginase has potential for chemoprevention of cancer and
should be evaluated in other models and in combination
with other drugs in anticipation of future clinical trials.
Table 3. Effect of inhibitors of PA metabolism on ODC activity in cells
of experimental Lewis lung carcinoma
Groups
Number
of ani-
mals
13th day 26th day
ODC activity,
A.U./mg pro-
tein
Inhi-
bition,
%
ODC activity,
A.U./mg protein
Inhi-
bition,
%
Control 4 0.034 ± 0.002 − 0.578 ± 0.070 −
nor-NOHA 5 0.070 ± 0.012# 0 0.346 ± 0.046* 40
DFMO 4 0.010 ± 0.001# 0 0.355 ± 0.060** 39
DFMO + nor-
NOHA 3 0.019 ± 0.001# 0 0.348 ± 0.050* 40
Note: #0.1 < p <0.25; *0.025 < p < 0.01; **0.01 < p < 0.005 as compared to control.
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| id | nasplib_isofts_kiev_ua-123456789-145472 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-11-28T13:54:39Z |
| publishDate | 2015 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Samoilenko, О.А. Milinevska, O.A. Karnaushenko, O.V. Shlyakhovenko, V.A. Zaletok, S.P. 2019-01-22T11:38:49Z 2019-01-22T11:38:49Z 2015 Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis / О.А. Samoilenko, O.A. Milinevska, O.V. Karnaushenko, V.A. Shlyakhovenko, S.P. Zaletok // Experimental Oncology. — 2015. — Т. 37, № 2. — С. 151-153. — Бібліогр.: 20 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/145472 Aim:To study the influence of polyamine metabolism inhibitors on the growth, metastasis and ornithine decarboxylase (ODC) activity of Lewis lung carcinoma. Materials and Methods: Experiments were performed on female mice C57Bl/6 with Lewis lung carcinoma. Nω-hydroxy-nor-arginine (nor-NOHA) and α-difluoromethylornithine (DFMO) were used as arginase and ODC inhibitors, correspondently. Inhibition of tumor growth was calculated by comparison of tumor volume in the treated and control groups. The average number of metastases per animal in the group and the average volume of pulmonary metastases per animal in the group have been determined. Determination of ODC — the key enzyme of the polyamine synthesis — in the samples of experimental tumors was performed by method of Luqman S. Results: Administration of DFMO or it’s combination with nor-NOHA resulted in the decrease of tumor growth rate, number and volume of lung metastases and was accompanied with reduced ODC activity in tumor tissue. Сonclusion: Modifiers of polyamine metabolism may be considered as promising targeted cancer therapy. Key Words: Lewis lung carcinoma, metastases, polyamine metabolism, ornithine decarboxylase, α-difluoromethylornithine, Nωhydroxy-nor-arginine. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Short communications Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis Article published earlier |
| spellingShingle | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis Samoilenko, О.А. Milinevska, O.A. Karnaushenko, O.V. Shlyakhovenko, V.A. Zaletok, S.P. Short communications |
| title | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis |
| title_full | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis |
| title_fullStr | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis |
| title_full_unstemmed | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis |
| title_short | Effect of polyamine metabolism inhibitors on Lewis lung carcinoma growth and metastasis |
| title_sort | effect of polyamine metabolism inhibitors on lewis lung carcinoma growth and metastasis |
| topic | Short communications |
| topic_facet | Short communications |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/145472 |
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