Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients
Aim: To investigate the effect of sodium humate on the level of cytogenetic damage in culture of lymphocytes of patients with thyroid cancer after γ-irradiation. Materials and Methods: Metaphase analysis of chromosome aberrations in cultured peripheral blood lymphocytes of 10 individuals with thyroi...
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nasplib_isofts_kiev_ua-123456789-1380032025-02-09T17:26:27Z Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients Shkarupa, V.M. Klymenko, S.V. Original contributions Aim: To investigate the effect of sodium humate on the level of cytogenetic damage in culture of lymphocytes of patients with thyroid cancer after γ-irradiation. Materials and Methods: Metaphase analysis of chromosome aberrations in cultured peripheral blood lymphocytes of 10 individuals with thyroid cancer was performed after irradiation of lymphocytes in vitro at a dose of 1 Gy from ¹³⁷Cs source at the early G₀ phase of cell cycle. Sodium humate was added to cell culture for 30 ± 15 min after phytohemagglutinin stimulation at concentrations of 10 and 100 μg/ml. Results: Sodium humate exhibited antimutagenic properties. The preparation at a concentration of 10 μg/ml was more effective than at a concentration of 100 μg/ml, reducing the average incidence of radiation-induced chromosome aberrations by 51.88 and 38.77%, respectively. The most pronounced antimutagenic effect of sodium humate was the reduction of the frequency of chromosomal type aberrations, however, such efficiency varied between individual patients with thyroid cancer. Conclusions: Sodium humate could be considered as a potential therapeutic modifier of radiation damage. 2016 Article Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients / V.M. Shkarupa, S.V. Klymenko // Experimental Oncology. — 2016 — Т. 38, № 2. — С. 108–111. — Бібліогр.: 22 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/138003 en Experimental Oncology application/pdf Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Original contributions Original contributions |
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Original contributions Original contributions Shkarupa, V.M. Klymenko, S.V. Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients Experimental Oncology |
| description |
Aim: To investigate the effect of sodium humate on the level of cytogenetic damage in culture of lymphocytes of patients with thyroid cancer after γ-irradiation. Materials and Methods: Metaphase analysis of chromosome aberrations in cultured peripheral blood lymphocytes of 10 individuals with thyroid cancer was performed after irradiation of lymphocytes in vitro at a dose of 1 Gy from ¹³⁷Cs source at the early G₀ phase of cell cycle. Sodium humate was added to cell culture for 30 ± 15 min after phytohemagglutinin stimulation at concentrations of 10 and 100 μg/ml. Results: Sodium humate exhibited antimutagenic properties. The preparation at a concentration of 10 μg/ml was more effective than at a concentration of 100 μg/ml, reducing the average incidence of radiation-induced chromosome aberrations by 51.88 and 38.77%, respectively. The most pronounced antimutagenic effect of sodium humate was the reduction of the frequency of chromosomal type aberrations, however, such efficiency varied between individual patients with thyroid cancer. Conclusions: Sodium humate could be considered as a potential therapeutic modifier of radiation damage. |
| format |
Article |
| author |
Shkarupa, V.M. Klymenko, S.V. |
| author_facet |
Shkarupa, V.M. Klymenko, S.V. |
| author_sort |
Shkarupa, V.M. |
| title |
Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients |
| title_short |
Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients |
| title_full |
Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients |
| title_fullStr |
Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients |
| title_full_unstemmed |
Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients |
| title_sort |
radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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2016 |
| topic_facet |
Original contributions |
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https://nasplib.isofts.kiev.ua/handle/123456789/138003 |
| citation_txt |
Radioprotective properties of sodium humate in radiation-induced mutagenesis in cultured lymphocytes of thyroid cancer patients / V.M. Shkarupa, S.V. Klymenko // Experimental Oncology. — 2016 — Т. 38, № 2. — С. 108–111. — Бібліогр.: 22 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
AT shkarupavm radioprotectivepropertiesofsodiumhumateinradiationinducedmutagenesisinculturedlymphocytesofthyroidcancerpatients AT klymenkosv radioprotectivepropertiesofsodiumhumateinradiationinducedmutagenesisinculturedlymphocytesofthyroidcancerpatients |
| first_indexed |
2025-11-28T15:57:00Z |
| last_indexed |
2025-11-28T15:57:00Z |
| _version_ |
1850050279506968576 |
| fulltext |
108 Experimental Oncology 38, 108–111, 2016 (June)
RADIOPROTECTIVE PROPERTIES OF SODIUM HUMATE
IN RADIATION-INDUCED MUTAGENESIS IN CULTURED
LYMPHOCYTES OF THYROID CANCER PATIENTS
V.M. Shkarupa*, S.V. Klymenko
State Institution “National Research Centre for Radiation Medicine of National Academy of Medical
Sciences of Ukraine”, Kyiv 04050, Ukraine
Aim: To investigate the effect of sodium humate on the level of cytogenetic damage in culture of lymphocytes of patients with thyroid
cancer after γ-irradiation. Materials and Methods: Metaphase analysis of chromosome aberrations in cultured peripheral blood
lymphocytes of 10 individuals with thyroid cancer was performed after irradiation of lymphocytes in vitro at a dose of 1 Gy from 137Cs
source at the early G0 phase of cell cycle. Sodium humate was added to cell culture for 30 ± 15 min after phytohemagglutinin stimula-
tion at concentrations of 10 and 100 μg/ml. Results: Sodium humate exhibited antimutagenic properties. The preparation at a con-
centration of 10 μg/ml was more effective than at a concentration of 100 μg/ml, reducing the average incidence of radiation-induced
chromosome aberrations by 51.88 and 38.77%, respectively. The most pronounced antimutagenic effect of sodium humate was the
reduction of the frequency of chromosomal type aberrations, however, such efficiency varied between individual patients with thyroid
cancer. Conclusions: Sodium humate could be considered as a potential therapeutic modifier of radiation damage.
Key Words: thyroid cancer, γ-irradiation, chromosome aberrations, sodium humate.
Currently, the scientific literature presents numer-
ous data on biological activity of humic substances.
Their preparations are being used both in medicine
and agriculture [1–4]. Humic substances are a com-
plex of natural polymer compounds, products of hu-
mification, which are present in the soil, peat, brown
coal, sapropel, etc. The main component of the humic
substances is humic acid, which physiologically active
form is salts (humates). The feature of humic sub-
stances is the saturation of their molecules by most
diverse functional groups, namely carboxyl, phenolic
and alcoholic hydroxyls, quinoid groups, methoxy, ami-
no and amido groups [1] that ultimately provides the
widest range of their biological activity [2–4]. Particular
attention is drawn to antitumor properties of humic
substances. Their antiangiogenic and proapoptotic
properties are found as well [5–7]. Antitumor activity
of humic substances was showed in various models
of transplantable tumors, spontaneous and induced
tumors, in human tumor cell lines [8]. Moreover, hu-
mic substances are characterized as non-toxic [4].
Noteworthy that preparations of humic substances
with antitumor activity do not have long-term adverse
effects [9].
In the last decade intense studies of humic sub-
stances have shown the potential of a significant
extending of their pharmacological application. The
scientific literature describes their antioxidant, im-
munostimulatory, anti-inflammatory and antiviral
effects [1–4, 10–12]. It was shown that humic sub-
stances may change the expression profile of more
than 30 genes, and affect DNA methylation [13, 14].
Considering adaptogenic and immune-stimulating
properties of humic substances, their ability to in-
crease non-specific resistance of a human body could
be assumed [1–4, 10–12]. There is an increasing in-
terest in the study of their use in cancer radiotherapy
to protect normal tissues, to improve the tolerability
of the treatment, and to reduce the risk of early and late
side effects. One of these effects of radiotherapy is its
significant mutagenic effect on non-transformed cells,
in particular of hematopoietic system, which increases
the risk of secondary cancer. It should be noted, that
most of studies testing this hypothesis were performed
in culture of lymphocytes of healthy individuals. There
are few publications describing studies of spontane-
ous and induced mutagenesis in diseased persons,
especially in cancer patients [15, 16].
Thyroid cancer is one of the most severe con-
sequences of the Chornobyl accident. Increased
quantity of TCR-mutant cells in patients with thyroid
cancer was revealed that may be due to genotoxic
effects, increased by genetic instability (especially
radiation-induced) [17]. In patients with thyroid cancer
living in areas contaminated with radionuclides, there
was found an increased level of cytogenetic damage
in cultured peripheral blood lymphocytes [15]. This
makes reasonable the attempts to modify radiation-
induced mutagenesis in somatic cells of patients with
thyroid cancer.
The aim of this work was to investigate the effect
of sodium humate on the level of cytogenetic damage
in cultured lymphocytes of patients with thyroid cancer
after γ-irradiation.
MATERIALS AND METHODS
Metaphase analysis of chromosomal aberrations
in peripheral blood lymphocyte cultures was carried
out in 10 thyroid cancer patients. The patients were
treated in National Research Center for Radiation
Medicine of National Academy of Medical Sciences
of Ukraine. The study has been performed in accor-
dance with ethics rules for biomedical research. All
Submitted: April 12, 2016.
*Correspondence: E-mail: Shkarupa_vlad@bigmir.net
Exp Oncol 2016
38, 2, 108–111
Experimental Oncology 38, 108–111, 2016 (June)38, 108–111, 2016 (June) (June) 109
patients gave an informed consent for the participa-
tion in the study.
Irradiation of lymphocytes was held in vitro
at a dose of 1 Gy from 137Cs source (IBL 437C, France,
dose rate — 2.46 Gy/min) in the early G0 phase of the
cell cycle. After phytohemagglutinin stimulation, dur-
ing 30 ± 15 min after radiation exposure, sodium hu-
mate (Agrohіmpak, Ukraine) was added to the culture
of lymphocytes at concentrations of 10 or 100 μg/ml.
In each case, at least 100 metaphases were examined
for numerical as well as structural aberrations. The
ave rage group indexes were calculated as the ratio
of the total number of chromosomal aberrations to the
total number of analyzed cells. Antimutagenic acti-
vity was determined by a reduction in the frequency
of chromosome aberrations that was calculated
as a percentage. Probability of differences was as-
sessed by the criterion of Fisher.
RESULTS AND DISCUSSION
Table 1 shows the results of analysis of the sodium
humate impact on the frequency of spontaneous and
radiation-induced chromosome aberrations in lympho-
cyte cultures of thyroid cancer patients.
Table 1. Antimutagenic effect of sodium humate in the culture of periphe-
ral blood lymphocytes of patients with thyroid cancer after irradiation in vi-
tro at a dose of 1 Gy from 137Cs
Treatment
Frequency
of chromosome
aberrations per
100 cells
Anti-
muta-
genic ef-
fect, %
Control 3.75 ± 0.51 –
Sodium humate, 10 μg/ml 2.61 ± 0.46* –
Sodium humate, 100 μg/ml 3.65 ± 0.52 –
Irradiation, 1 Gy 21.51 ± 0.95 –
Irradiation, 1 Gy + sodium humate, 10 μg/ml 10.35 ± 0.66** 51.88***
Irradiation, 1 Gy + sodium humate, 100 μg/ml 13.17 ± 0.69** 38.77
Note: *р = 0.08 compared to untreated control; **p < 0.05 compared to the
effect of irradiation; ***р < 0.05 compared to irradiation, 1 Gy with addition
of sodium humate, 100 μg/ml.
The average spontaneous frequency of chro-
mosome aberrations was 3.75 ± 0.51 aberrations
per 100 cells. However, a range of minimal and
maximal individual values of this index varied from
0.00 to 8.15 ± 2.36 aberrations per 100 cells. Sodium
humate at a concentration of 100 μg/ml did not reduce
the mean values of spontaneous mutagenesis. The
tendency to reduce the average frequency of sponta-
neous aberrations in cultured lymphocytes of thyroid
cancer patients (2.61 ± 0.46 aberrations/100 cells)
modified by humic preparation at a concentration
of 10 μg/ml had insufficient level of statistical signifi-
cance (p = 0.08). The average frequency of chromo-
some aberrations in cultured lymphocytes of patients
with thyroid cancer after irradiation in vitro was 21.51 ±
0.95 aberrations/100 cells. The range of minimal and
maximal individual values of this index varied from
13.16 ± 2.49 to 27.50 ± 4.08 aberrations per 100 cells.
Addition of sodium humate after exposure has shown
an expressed antimutagenic effect. Antimutagenic
efficiency of sodium humate at a concentration
of 10 μg/ml (antimutagenic effect — 51.88%) was
significantly higher (p < 0.05) than by the impact of the
preparation at a concentration of 100 μg/ml (antimu-
tagenic effect — 38.77%).
Reduction of the radiation-induced chromosome
aberrations frequency as a result of the sodium humate
action was mainly due to the decrease of aberrations
of chromosomal type (Table 2). Reduction of the
average frequency of aberrations of chromatide type
to 26.44% by addition of the preparation at a concen-
tration of 10 μg/ml, and to 24.92% by addition of the
preparation at a concentration of 100 μg/ml had not
reached statistical significance.
Table 2. Effect of sodium humate on the frequency of radiation-induced
chromosomal and chromatide type aberrations (average value)
Treatment
Frequen-
cy of chroma-
tide type aber-
rations per
100 cells
Anti-
muta-
genic
effect,
%
Frequen-
cy of chromo-
somal type
aber rations
per 100 cells
Anti-
muta-
genic
effect,
%
Irradiation, 1 Gy 3.29 ± 0.41 – 18.22 ± 0.90 –
Irradiation, 1 Gy + sodi-
um humate, 10 μg/ml
2.42 ± 0.33 26.44 7.92 ± 0.58* 56.53*
Irradiation, 1 Gy + sodi-
um humate, 100 μg/ml
2.47 ± 0.32 24.92 10.70 ± 0.63* 41.27*
Note: *p < 0.05 as compared to the effect of irradiation.
Analysis of cytogenetic indexes at the individual
level (Table 3 and 4) found significant differences in the
response on radiation exposure and antimutagenic
effect of sodium humate.
Table 3. Individual indexes of antimutagenic effect of sodium humate
(10 μg/ml) in the culture of peripheral blood lymphocytes of patients with
thyroid cancer after irradiation in vitro at a dose of 1 Gy from 137Cs
Culture
of lym-
phocytes,
pa-
tient’s ID
Frequency of chromosome aberrations per 100 cells Anti-
muta-
genic
effect,
%
Control
Sodium hu-
mate
(10 μg/ml)
Irradiation,
1 Gy
Irradiation,
1 Gy + sodium
humate
(10 μg/ml)
Т1 0.00 ± 0.00 1.00 ± 0.99 18.00 ± 2.72 9.55 ± 1.98** 46.94
Т2 0.39 ± 0.38 0.83 ± 0.83 13.16 ± 2.49 4.76 ± 1.47** 63.83
Т3 0.73 ± 0.72 0.95 ± 0.95 16.33 ± 2.13 6.67 ± 1.44** 59.16
Т4 1.00 ± 0.99 2.00 ± 1.40 22.50 ± 3.30 16.67 ± 2.54*** 25.91
Т5 2.99 ± 1.47 2.00 ± 1.40 19.00 ± 2.77 8.00 ± 1.92** 57.90
Т6 3.85 ± 1.69 2.00 ± 1.40 16.00 ± 2.99 6.00 ± 1.37** 62.50
Т7 4.88 ± 1.51 3.00 ± 1.71 19.00 ± 3.92 16.00 ± 3.67*** 15.79
Т8 5.80 ± 1.56 5.00 ± 1.62 15.00 ± 3.57 14.00 ± 2.45*** 6.67
Т9 6.60 ± 1.46 2.73 ± 1.09* 27.25 ± 2.09 10.00 ± 2.12** 63.30
Т10 8.15 ± 2.36 6.00 ± 2.38 27.50 ± 4.08 18.50 ± 2.75** 32.73
Note: *р < 0.05 compared to untreated control; **p < 0.05 compared to the
effect of irradiation; ***р > 0.05 compared to the effect of irradiation.
Table 4. Individual indexes of antimutagenic effects of sodium humate
(100 μg/ml) in the culture of peripheral blood lymphocytes of patients with
thyroid cancer after irradiation in vitro at a dose of 1 Gy 137Cs
Culture
of lym-
phocytes,
pati-
ent’s ID
Frequency of chromosome aberrations per 100 cells Anti-
muta-
genic
effect,
%
Control
Sodium
humate
(100 μg/ml)
Irradiation,
1 Gy
Irradiation,
1 Gy + sodi-
um humate
(100 μg/ml)
Т1 0.00 ± 0.00 1.00 ± 0.99 18.00 ± 2.72 10.00 ± 2.02** 44.44
Т2 0.39 ± 0.38 0.87 ± 0.87 13.16 ± 2.49 5.58 ± 1.64** 57.60
Т3 0.73 ± 0.72 1.82 ± 1.28 16.33 ± 2.13 10.00 ± 2.37** 38.76
Т4 1.00 ± 0.99 3.33 ± 1.79 22.50 ± 3.30 15.83 ± 3.33*** 29.64
Т5 2.99 ± 1.47 4.00 ± 1.96 19.00 ± 2.77 9.50 ± 2.07** 50.00
Т6 3.85 ± 1.69 4.17 ± 1.82 16.00 ± 2.99 6.00 ± 1.37** 62.50
Т7 4.88 ± 1.51 4.00 ± 1.96 19.00 ± 3.92 15.83 ± 3.33*** 16.68
Т8 5.80 ± 1.56 6.79 ± 1.98 15.00 ± 3.57 17.60 ± 1.46*** −17.33
Т9 6.60 ± 1.46 3.79 ± 1.18* 27.25 ± 2.09 16.89 ± 2.50** 38.02
Т10 8.15 ± 2.36 7.00 ± 2.55 27.50 ± 4.08 18.45 ± 2.70 32.91
Note:*р < 0.05 compared to untreated control; **p < 0.05 compared to the
effect of irradiation; ***р > 0.05 compared to the effect of irradiation.
Most researchers believe that if the preparation
reduces the induced damage by less than 10%,
110 Experimental Oncology 38, 108–111, 2016 (June)
it could not be considered antimutagenic. Substances
that reduce the frequency of aberrations at the range
of 10.1–20.0% usually are being called “very weak”,
and at in the range of 20.1–40.0% — “weak” anti-
mutagens. Reducing the frequency of aberrations
by 40.1–80.0% is typical for the majority of antimu-
tagens of “average” performance. Substances, that
can reduce the level of induced damage by more
than 80%, are related to the group of “strong” anti-
mutagens [18]. In cultures of lymphocytes of patients
T7 and T8 sodium humate did not show antimutagenic
properties. In a lymphocyte culture of patient T4 weak
antimutagenic activity of humic preparation (25.91%
at a concentration of 10 μg/ml and 29.64% at a con-
centration of 100 μg/ml) had insufficient statistical
significance. In other cases, sodium humate has
showed the properties of antimutagen of “average
efficiency”. Antimutagenic effect of sodium humate
did not depend on the level of spontaneous or initial
radiation-induced mutagenesis.
In the context of possible modifying of the spon-
taneous and induced mutagenesis in the culture
of human lymphocytes the study on radioprotective
properties of timalin should be mentioned [15, 16].
In the culture of lymphocytes of thyroid cancer pa-
tients, who reside in the territories of Ukraine with
increased density of radioactive contamination from
the Chornobyl accident, timalin reduced the average
level of spontaneous chromosome aberrations from
6.0 ± 0.7 to 2.0 ± 0.5% [15]. In our study, the aver-
age frequency of chromosome aberrations induced
by sodium humate at a concentration of 10 μg/ml was
similar and amounted to 2.61 ± 0.46 aberrations
per 100 cells, although the spontaneous frequency
was lower and amounted to 3.75 ± 0.51 aberrations
per 100 cells. It is possible that studied substances
can reduce the frequency of spontaneous chromo-
some aberrations only to a certain level, or under
particular circumstances of increased spontaneous
mutagenesis.
In a situation, where the individual frequency
of spontaneous chromosome aberrations in the cul-
tured lymphocytes of thyroid cancer patients was with-
in the average population values (0–3 aberrations per
100 cells), it showed no reduction of spontaneous level
by addition of sodium humate. Among those with high
levels of spontaneous mutagenesis, antimutagenic
effect of sodium humate manifested in lymphocyte
cultures of two patients (T9 and T10), in one of which
(T9) preparation in both studied concentrations leads
to the significant reduction of spontaneous frequency
of chromosome aberrations.
Dyomina [15] have shown that the timalin added
to cultured lymphocytes of thyroid cancer patients
irradiated at a dose of 0.3 Gy, reduced the percent-
age of aberrant cells from 9.0 ± 1.7 to 2.1 ± 1.0%,
compared to irradiated control. So, timalin could
be effective if added before irradiation [19]. During
timalin action on lymphocytes at G1 phase of cell
cycle subsequently exposed to a high dose irradiation
(2 Gy), the author observed an opposite cytogenetic
effect, an increased frequency of radiation-induced
aberrations from 18.0 ± 1.9 to 26.0 ± 2.5%, which was
explained by timalin influence on radiation-induced
mitotic delay [16]. The essential difference between
these data and our study was that sodium humate
showed antimutagenic effectiveness by its action after
irradiation exposure. This allows considering it as a po-
tential therapeutic modifier of radiation damage [19].
It is believed that effectiveness of radioprotec-
tive substances after cell irradiation could be caused
by activation of DNA repair or other irradiation reco-
very processes, in particular stimulation of repopula-
tion [19]. Repopulation as a reaction of organisms
to irradiation exposure was studied in detail in many
model systems, including its stimulation by antimu-
tagens [19–21]. Well-known adaptogenic proper-
ties of humates suppose their impact also on other
processes of post-radiation recovery [1–4]. The view
on mechanisms of radioprotective action of sodium hu-
mate in the culture of human lymphocytes is supported
by the results of our previous studies done in plant
test-system. In particular, the multiplicity of genopro-
tective mechanisms of humic substances, including
the activating reparation processes and stimulation
of repopulation has been shown [22].
In conclusion, antimutagenic effect of sodium
humate was revealed, what allows to consider it as po-
tential therapeutic modifier of radiation damage. The
preparation at a concentration of 10 μg/ml was more
effective than at a concentration of 100 μg/ml, re-
ducing the average incidence of radiation-induced
chromosome aberrations by 51.88 and 38.77%,
respectively. Antimutagenic effect was manifested
mainly by reduction of the frequency of chromosome
type aberrations. There were individual differences
in antimutagenic efficacy of sodium humate in cultured
lymphocytes of thyroid cancer patients.
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