Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice
Aim: It has been shown previously that chronic social defeat stress produces development ofstrong anxiety and increases intensity of experimental metastasis in the losers in comparison with the winners and control mice. The question was: isit possible to decrease the number ofmetastases inthe lose...
Збережено в:
| Дата: | 2009 |
|---|---|
| Автори: | , , , , , |
| Формат: | Стаття |
| Мова: | English |
| Опубліковано: |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2009
|
| Назва видання: | Experimental Oncology |
| Теми: | |
| Онлайн доступ: | http://dspace.nbuv.gov.ua/handle/123456789/135107 |
| Теги: |
Додати тег
Немає тегів, Будьте першим, хто поставить тег для цього запису!
|
| Назва журналу: | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| Цитувати: | Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice / V.I. Kaledin, S.I. llnitskaya, V.P. Nikolin, N.A. Popova, D.A. Smagin, N.N. Kudryavtseva // Experimental Oncology. — 2009. — Т. 31, № 1. — С. 62–64. — Бібліогр.: 18 назв. — англ. |
Репозитарії
Digital Library of Periodicals of National Academy of Sciences of Ukraine| id |
irk-123456789-135107 |
|---|---|
| record_format |
dspace |
| spelling |
irk-123456789-1351072018-06-15T03:10:17Z Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice Kaledin, V.I. llnitskaya, S.I. Nikolin, V.P. Popova, N.A. Smagin, D.A. Kudryavtseva, N.N. Short communications Aim: It has been shown previously that chronic social defeat stress produces development ofstrong anxiety and increases intensity of experimental metastasis in the losers in comparison with the winners and control mice. The question was: isit possible to decrease the number ofmetastases inthe losers by chronic or acute diazepam treatment. Materials and Methods: Sensory contact model was used for generating male mice with repeated experience of social victories or defeats in daily agonistic interactions. Tumor cells of Lewis Lung Carcinoma (LLC) were injected into the tail vein of animals after 10 days of agonistic interactions. Then mice were treated acutely or chronically (7 days) with diazepam (1 mg/kg, i. p). Number ofmetastases in the lung was calculated in 16 days after tumor cell transplantation. Results: Diazepam decreased the number of LLC metastases in anxious losers, whereas in the winners and control mice, without anxiety state, diazepam was ineffective. Conclusion: Well-known anxiolytic diazepam may decrease intensity of metastasis in anxious mice. 2009 Article Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice / V.I. Kaledin, S.I. llnitskaya, V.P. Nikolin, N.A. Popova, D.A. Smagin, N.N. Kudryavtseva // Experimental Oncology. — 2009. — Т. 31, № 1. — С. 62–64. — Бібліогр.: 18 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/135107 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| institution |
Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Short communications Short communications |
| spellingShingle |
Short communications Short communications Kaledin, V.I. llnitskaya, S.I. Nikolin, V.P. Popova, N.A. Smagin, D.A. Kudryavtseva, N.N. Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice Experimental Oncology |
| description |
Aim: It has been shown previously that chronic social defeat stress produces development ofstrong anxiety and increases intensity of experimental
metastasis in the losers in comparison with the winners and control mice. The question was: isit possible to decrease the number
ofmetastases inthe losers by chronic or acute diazepam treatment. Materials and Methods: Sensory contact model was used for generating
male mice with repeated experience of social victories or defeats in daily agonistic interactions. Tumor cells of Lewis Lung Carcinoma
(LLC) were injected into the tail vein of animals after 10 days of agonistic interactions. Then mice were treated acutely or chronically
(7 days) with diazepam (1 mg/kg, i. p). Number ofmetastases in the lung was calculated in 16 days after tumor cell transplantation. Results:
Diazepam decreased the number of LLC metastases in anxious losers, whereas in the winners and control mice, without anxiety state,
diazepam was ineffective. Conclusion: Well-known anxiolytic diazepam may decrease intensity of metastasis in anxious mice. |
| format |
Article |
| author |
Kaledin, V.I. llnitskaya, S.I. Nikolin, V.P. Popova, N.A. Smagin, D.A. Kudryavtseva, N.N. |
| author_facet |
Kaledin, V.I. llnitskaya, S.I. Nikolin, V.P. Popova, N.A. Smagin, D.A. Kudryavtseva, N.N. |
| author_sort |
Kaledin, V.I. |
| title |
Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice |
| title_short |
Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice |
| title_full |
Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice |
| title_fullStr |
Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice |
| title_full_unstemmed |
Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice |
| title_sort |
limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2009 |
| topic_facet |
Short communications |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/135107 |
| citation_txt |
Limiting effect of diazepam on lewis lung carcinoma metastasis in anxious male mice / V.I. Kaledin, S.I. llnitskaya, V.P. Nikolin, N.A. Popova, D.A. Smagin, N.N. Kudryavtseva // Experimental Oncology. — 2009. — Т. 31, № 1. — С. 62–64. — Бібліогр.: 18 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
AT kaledinvi limitingeffectofdiazepamonlewislungcarcinomametastasisinanxiousmalemice AT llnitskayasi limitingeffectofdiazepamonlewislungcarcinomametastasisinanxiousmalemice AT nikolinvp limitingeffectofdiazepamonlewislungcarcinomametastasisinanxiousmalemice AT popovana limitingeffectofdiazepamonlewislungcarcinomametastasisinanxiousmalemice AT smaginda limitingeffectofdiazepamonlewislungcarcinomametastasisinanxiousmalemice AT kudryavtsevann limitingeffectofdiazepamonlewislungcarcinomametastasisinanxiousmalemice |
| first_indexed |
2025-07-09T22:43:15Z |
| last_indexed |
2025-07-09T22:43:15Z |
| _version_ |
1837211057275273216 |
| fulltext |
62 Experimental Oncology 31, 62–64, 2009 (March)
It has been experimentally shown that chronic stress
of different etiology is the factor provoking a rapid tumor
growth and metastasis [1–7]. Our data obtained earlier
indicated expressed and replicable effects of the so-
cial status of mice on experimental metastasis of two
transplantable tumors Lewis lung carcinoma (LLC) and
Hepatocarcinoma-29: in the losers with repeated experi-
ence of social defeats the number of metastases in the
lungs was significantly higher than that in the winners with
repeated social victories or control mice [8, 9]. Winners
and losers are considered to be animals with positive
or negative emotional states, respectively [10]. In has
been shown the development of strong anxiety in the los-
ers under social defeat stress [11–13]. Anxious behavior
in the losers was sensitive to anxiolytic drugs: chronic [14]
but not acute treatment [14, 15] by different anxiolytics
had positive effects on losers’ psychoemotional state.
Question was: does the intensity of experimental me-
tastasis depend on duration of agonistic interactions and
could pharmacological correction of psycho emotional
state decrease the number of metastases in anxious
animals. It this experiment we studied animals after ten
agonistic interactions because the losers, but not the win-
ners, demonstrated high level of anxiety estimated in dif-
ferent behavioral tests (plus maze, partition tests etc)
in comparison with controls [11]. Anxiolytic diazepam,
which is shown to be active in this model [16] was used.
The intensity of experimental metastasis was studied
after chronic and acute injections of diazepam.
Experimental animals. Mice of C57BL/6J strain
were bred and kept under the standard vivar ium con-
ditions at the Institute of Cytology and Genet ics, Si-
berian Division, Russian Academy of Sciences. Mice
were housed on a 12 h/12 h light/dark regimen and
received the standard food (pellets) and water ad libi-
tum. Weaned at the age of one month, the males were
housed before the experiment in groups of 8–10 indivi-
duals in 36 x 23 x 12 cm cages. Mice used in experiments
were 10–12 weeks old. All experi mental procedures were
in compliance with the Euro pean Communities Council
Directive of November 24, 1986 (86/609/EEC).
Technique for generation of male mice with re-
peated experience of agonistic interactions. Win-
ners and losers were generated using the sensory con-
tact model [10]. Pairs of animals were placed in steel
cages (28 x 14 x 10 cm) divided in two compartments
by a perforated transparent partition allowing the
animals to see, hear and smell their neighbor, but not
to contact them physically. Test sessions commenced
2 days after adaptation of the animals to these new
housing conditions (sensory contact). Every afternoon
(between 2.00 p. m. and 5.00 p. m.) the steel cover
of the cage was replaced by a transparent one, and
5 min later (the period needed for adaptation to the
lighting condition) the partition was removed for 10 min
to allow agonistic interactions. Superiority of one of the
partners was evident within 2–3 daily test sessions.
One partner attacked, bit, and chased the other, who
displayed defensive behavior only (sideways, upright
postures, withdrawal, lying on the back or freezing).
Agonistic interactions were discontinued by lowering
the partition if intensive attacks lasted more than 3 min.
Every day after the test session, each defeated mouse
was placed in another two compartment cage with
a partition, in which another winner was present in the
other compartment. The winners remained in their
own compartments. The procedure yielded equal
numbers of males with an opposite social experience
of aggression, evidenced by victories (aggressors,
winners) and defeats (defeated mice, losers) in ago-
nistic interactions. In this experi ment, the control group
was represented by the males that lived together for
a long time in groups with stable dominant-subordinate
relationships. It was shown that, irrespective of the so-
LIMITING EFFECT OF DIAZEPAM ON LEWIS LUNG CARCINOMA
METASTASIS IN ANXIOUS MALE MICE
V.I. Kaledin, S.I. llnitskaya, V.P. Nikolin, N.A. Popova, D.A. Smagin, N.N. Kudryavtseva*
Institute of Cytology and Genetics, Siberian Department of the Russian Academy of Sciences, Novosibirsk
630090, Russia
Aim: It has been shown previously that chronic social defeat stress produces development of strong anxiety and increases intensity of ex-
perimental metastasis in the losers in comparison with the winners and control mice. The question was: is it possible to decrease the number
of metastases in the losers by chronic or acute diazepam treatment. Materials and Methods: Sensory contact model was used for generating
male mice with repeated experience of social victories or defeats in daily agonistic interactions. Tumor cells of Lewis Lung Carcinoma
(LLC) were injected into the tail vein of animals after 10 days of agonistic interactions. Then mice were treated acutely or chronically
(7 days) with diazepam (1 mg/kg, i. p). Number of metastases in the lung was calculated in 16 days after tumor cell transplantation. Results:
Diazepam decreased the number of LLC metastases in anxious losers, whereas in the winners and control mice, without anxiety state,
diazepam was ineffective. Conclusion: Well-known anxiolytic diazepam may decrease intensity of metastasis in anxious mice.
Key Words: agonistic interactions, anxiety, Lewis lung carcinoma, metastasis, diazepam.
Received: July 7, 2008.
*Correspondence: Fax: 73833331278
E-mail: natnik@bionet.nsc.ru
Abbreviations used: LLC — Lewis lung carcinoma; T10 winners,
T10 losers — animals after 10 days of agonistic interactions; T20 win-
ners, T20 losers — animals after 20 days of agonistic interactions.
Exp Oncol 2009
31, 1, 62–64
Experimental Oncology 31, 62–64, 2009 (March) 63
cial status (dominant or subordinate), animals in such
groups are not stressed [17].
Tumor model and animal’s treatment. First
we studied experimental metastasis of Lewis lung
carcinoma (LLC) in male mice after 10 and 20 days
of agonistic interactions (T10 and T20 losers and
T10 and T20 winners). LLC was maintained by i.m. pas-
sages on C57BL/6J mice. To produce the tumor cell
inocula, tumor transplants were removed, cut up with
scissors; the suspension was filtered through a stain-
less steel mesh and spun two times at 500 g. The pellet
was washed in physiological solution and resuspended
to a final concentration of 5 x 105 cell/mL. LLC was
transplanted to mice via i. v. injections (0.5 ml of cell
suspension) into tail vein. In each experiment, the
tumor cells were grafted simultaneously to mice of all
groups: the winners, losers and controls. After tumor
transplantation, the animals were kept in their com-
partments without agonistic interactions for 16 days
until the end of the experiment. On day 17, the mice
were decapitated; the lungs were fixed in 10% forma-
lin and tested for the presence of metastases using
a binocular magnifier (magnification 8 ×).
Chronic and acute injections of diazepam. After
cell inoculation, each group of T10 winners and T10 los-
ers as well as control mice were divided into two sub-
groups. Beginning the day after tumor transplantation,
the mice of one of the subgroups received 7 daily i. p.
injections of diazepam (Polfa Tarchomin S.A.) at dose
of 1 mg/kg, and the animals of every other subgroup
received saline (10 ml/kg body weight) in the same way.
The experiment was terminated on 17 days after tumor
challenge and the number of metastases in the lungs
was calculated. Similar experimental design was used
to study effect of acute injection of diazepam: beginning
the day after tumor transplantation, the mice of one
of the subgroups received acute injection of diazepam
(1 mg/kg), and the animals of every other subgroup
received acutely saline. Experiment was terminated
on 17 days after tumor challenge and the number
of metastases in the lungs was calculated.
Statistical analysis. Number of metastases was
statistically processed using one-way ANOVA with the
factor “social group” (winners, losers, control) for ani-
mals with T10 and T20 days of agonistic interactions.
Two-way ANOVA was used for experiments with phar-
macological treatment with the factor “social group”
(winners, losers, control), factor “treatment” (saline,
diazepam) and interaction of these factors. Then,
paired comparisons of groups were performed using
Student’s t-test. Differences between experimental
groups were considered to be significant if p < 0.05.
The data on the number of experimental lung metas-
tases in the control group, winners and losers inoculated
i.v. with tumor cells after 10 and 20 days of agonistic in-
teractions are presented in Fig 1, a, b. One-way ANOVA
showed that the number of meta stases was reliably
determined by the social group of animals with 10 days
of agonistic interactions [F(2,34) = 4.0; p < 0.027]
and 20 days of agonistic interactions [F(2.68) = 6.85;
p < 0.002]. Comparisons of the data for mice after
10 days of social expe rience using Student’s t-test did
not show differences in the number of metastases be-
tween the control and T10 winners (p > 0.05). However,
the T10 winners vs T10 losers, as well as the control
mice vs T10 lo sers differed significantly by this param-
eter (for both comparisons p < 0.01). No differences
were found in the number of metastases between the
T20 winners and control mice (p > 0.05), whereas the
T20 losers and T20 winners as well as the T20 losers
and control mice differed significantly (p ≤ 0.01 and
p ≤ 0.001, respectively). Thus, the data obtained are
in agreement with our previous findings [8, 9]. Moreover
the intensity of experimental metastasis in the losers de-
pends on duration of agonistic interactions. The number
of metastasis was about 2,6 times more in T20 losers
and about 2.0 times more in T10 losers in comparison
with the respective controls.
0
2
4
6
8
10
Control Winners Losers
# *
M
et
as
ta
se
s,
N
M
et
as
ta
se
s,
N
0
2
4
6
8
10
Control Winners Losers
*#
a
b
Fig. 1. Number of LLC metastases in lung of mice with repeated
social defeats (losers), repeated social victories (winners) and
the control after 10 (a) and 20 (b) days of agonistic interactions.
*p < 0.01; vs Control; #p < 0.01; vs Winners.
Chronic diazepam injections during 7 days had limi-
ting effects on metastasis in T10 losers. Two-way ANO-
VA showed that the number of metastases was reliably
determined by the social group of animals [F(2,61) =
3.32; p < 0.05], and interaction between effects of dia-
ze pam and groups of animals [F(2.61) = 3.63; p < 0.05]
was found. In diazepam-treated losers, the number
of metastases is decreased significantly in comparison
with saline-treated losers (p < 0.05) and was similar
with the metastasis levels in the winners and control
animals. In the winners and control mice, however, dia-
zepam neither potentiated nor inhibited lung metastasis
(p > 0.05) (Fig. 2, b). Acute injection of dia zepam was
not too effective in decreasing the number of metastasis
in the losers: there were no influence of interactions
64 Experimental Oncology 31, 62–64, 2009 (March)
between effects of diazepam and groups of animals
[F(2.61) = 0.54; p > 0.05] (Fig. 2, a).
a
b
0
2
4
6
8
10
12
Control Winners Losers
*
M
et
as
ta
se
s,
N
M
et
as
ta
se
s,
N
0
5
10
15
20
25
Control Winners Losers
*
#
Fig. 2. Number of LLC metastases in lung of mice with repeated
social defeats (losers), repeated social victories (winners) and
the control after acute (a) and chronic (b) diazepam treatment.
Light column — saline injections; dark columns — diazepam
injections. *p < 0.05 vs Control and Winners; #p < 0.05 vs saline-
treated Losers.
Noteworthy, T10 losers, but not T10 winners, had
high level of anxiety in comparison with control [11] and
diazepam specifically decreased the number of me-
tastases only in anxious mice, defeated losers. It has
been shown recently [18] that other drug — ethanol
chronically injected during 7 days starting with day
of tumor cells injections also decreased the number
of lung metastases in T20 losers. Since ethanol has
been demonstrated to have anxiolytic effects [13], these
results are in agreement with effects of dia zepam. Thus,
it may be supposed that pharmacological correction
of psychoemotional state with anxiolytics may decrease
the intensity of metastasis in anxious individuals.
REFERENCES
Azpiroz A, De Miguel Z, Fano E, Vegas O. 1. Relations
between different coping strategies for social stress, tumor
development and neuroendocrine and immune activity in male
mice. Brain Behav Immun 2008; 22: 690–8.
Azpiroz A, Garmendia L, Fano E, Sanches-Martin JR. 2.
Relation between aggressive behavior, immune activity, and
disease susceptibility. Aggress Violent Behav 2003; 8: 433–53.
Giraldi T, Perissin L, Zorzet S, 3. et al. Metastasis and
neuroendocrine system in stressed mice. Int J Neurosci 1994;
74: 265–78.
Ishihara Y, Matsunaga K, Iijima H, 4. et al. Time-depen-
dent effects of stressor application on metastasis of tumor cells
in the lung and its regulation by an immunomodulator in mice.
Psychoneuroendocrinol 1999; 24: 713–26.
Palermo-Neto J, de Oliveira Massoco C, Robespierre 5.
de Souza W. Effects of physical and psychological stressors
on behavior, macrophage activity, and Ehrlich tumor growth.
Brain Behav Immun 2003; 17: 43–54.
Stefanski V, Ben-Eliyahu S. 6. Social confrontation and
tumor metastasis in rats: defeat and beta-adrenergic mecha-
nism. Physiol Behav 1996; 60: 277–82.
Vegas O, Beitia G, Sanchez-Martin JR, 7. et al. Behavioral
and neurochemical responses in mice bearing tumors submit-
ted to social stress. Behav Brain Res 2004; 155: 125–34.
Kaledin VI, Tenditnik MV, Nikolin VP, 8. et al. Effect
of psychoemotional state on growth and metastasis of Lewis tu-
mor in mice. Dokl Acad Sci 2006; 406: 57–9 (In Russian).
Kudryavtseva NN, Tenditnik MV, Nikolin VP, et al. 9. The
influence of psychoemotional status on metastasis of Lewis Lung
Carcinoma and Hepatocarcinoma-29 in mice of C57BL/6J and
CBA/LAC strains. Exp Oncol 2007; 29: 35–8.
Kudryavtseva NN. 10. Agonistic behavior: a model, ex-
perimental studies, and perspectives. Neurosci Behav Physiol
2000; 30: 293–305.
Avgustinovich DF, Gorbach OV, Kudryavtseva NN. 11.
Comparative analysis of anxiety-like behavior in partition and
plus-maze tests after agonistic interactions in mice. Physiol
Behav 1997; 61: 37–43.
Kudryavtseva NN, Avgustinovich DF. 12. Behavioral and
physiological markers of experimental depression induced
by social conflicts (DISC). Aggr Behav 1998; 24: 271–86.
Kudryavtseva NN, Gerrits MAFM, Avgustinovich DF, 13.
et al. Anxiety and ethanol consumption in victorious and
defeated mice: effect of k-opioid receptor activation. Eur
Neuropsychopharmacology 2006; 16: 504–11.
Kudryavtseva NN, Avgustinovich DF, Bondar NP, 14. et al.
An experimental approach for the study of psychotropic drug
effects under simulated clinical conditions. Curr Drug Metab
2008; 9: 4: 352–60.
Rygula R, Abumaria N, Havemann-Reinecke U, 15. et al.
Pharmacological validation of a chronic social stress model
of depression in rats: effects of reboxetine, haloperidol and
diazepam. Behav Pharmacol 2008; 19: 183–96.
Amikishieva AV, Semendyaeva SN. 16. Psychotropic effects
of diazepam in male mice with alternative psychoemotional
states. Russ Physiol J 2006; 92: 488–98 (In Russian).
Bartolomucci A, Palanza P, Parmigiani S. 17. Group
housed mice: are they really stressed? Aggress Violent Behav
2003; 14: 341–50.
Ilnitskaya SI, Nikolin VP, Popova NA, 18. et al. Effect
of ethanol on metastasis of Lewis lung carcinoma in male
mice with different emotional. Ross Physiol J im. Seche-
nova IM 2008; 95: 1: 74–8.
Copyright © Experimental Oncology, 2009
Подп. в печать 11.03.2009. Формат 60×841/8. Печать офс. Бумага офс. Усл. печ. л. 7,44. Уч.–изд. л. 8,33. Тираж 300 экз. Зак. № 29-4730.
Отпечатано с готовых фотоформ в издательско-полиграфическом центре “Киевский университет”, 01033, Киев, бульв. Т. Шевченко, 14.
|