Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue
Aim: The aim of this study was to evaluate the frequency of existence of thyroid extratumoral normo- and microfollicular tissue in patients with thyroid carcinoma and peculiarities of apoptosis in mentioned tissue. Materials and Methods: Using samples of normo- and microfollicular thyroid tissue it...
Збережено в:
| Дата: | 2012 |
|---|---|
| Автори: | , , |
| Формат: | Стаття |
| Мова: | English |
| Опубліковано: |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2012
|
| Назва видання: | Experimental Oncology |
| Теми: | |
| Онлайн доступ: | http://dspace.nbuv.gov.ua/handle/123456789/138690 |
| 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: | Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue / T.M. Myshunina, O.V. Kalinichenko, N.D. Tronko // Experimental Oncology. — 2012. — Т. 34, № 2. — С. 112-115. — Бібліогр.: 21 назв. — англ. |
Репозитарії
Digital Library of Periodicals of National Academy of Sciences of Ukraine| id |
irk-123456789-138690 |
|---|---|
| record_format |
dspace |
| spelling |
irk-123456789-1386902018-06-20T03:07:26Z Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue Myshunina, T.M. Kalinichenko, O.V. Tronko, N.D. Original contributions Aim: The aim of this study was to evaluate the frequency of existence of thyroid extratumoral normo- and microfollicular tissue in patients with thyroid carcinoma and peculiarities of apoptosis in mentioned tissue. Materials and Methods: Using samples of normo- and microfollicular thyroid tissue it was determined the content of fragmented DNA and intensity of stimulated internucleosomal DNA fragmentation; activities of caspase-3 and cysteine lysosomal cathepsins. Results: It was found that normofollicular tissue is observed more often in patients with nodal euthyroid goiter but microfollicular tissue is more common for patients with carcinoma. Extratumoral microfollicular tissue was found in the thyroid of patients above 50 years old mostly, and more rarely in young ones. The fragmented DNA concentration in microfollicular tissue was lower by a factor of 3.5 and intensity of stimulated internucleosomal DNA fragmentation was also decreased. Activity both of cathepsin B in lisosomes and caspase-3 in lysates of such tissue was also decreased. Conclusions: The decrease of intensity of spontaneous apoptosis and the absence of its modulation/induction following proapoptotic factors in extratumoral microfollicular thyroid tissue may be considered as a respond of the thyroid gland tissue to an existence of carcinoma. 2012 Article Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue / T.M. Myshunina, O.V. Kalinichenko, N.D. Tronko // Experimental Oncology. — 2012. — Т. 34, № 2. — С. 112-115. — Бібліогр.: 21 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/138690 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 |
Original contributions Original contributions |
| spellingShingle |
Original contributions Original contributions Myshunina, T.M. Kalinichenko, O.V. Tronko, N.D. Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue Experimental Oncology |
| description |
Aim: The aim of this study was to evaluate the frequency of existence of thyroid extratumoral normo- and microfollicular tissue in patients with thyroid carcinoma and peculiarities of apoptosis in mentioned tissue. Materials and Methods: Using samples of normo- and microfollicular thyroid tissue it was determined the content of fragmented DNA and intensity of stimulated internucleosomal DNA fragmentation; activities of caspase-3 and cysteine lysosomal cathepsins. Results: It was found that normofollicular tissue is observed more often in patients with nodal euthyroid goiter but microfollicular tissue is more common for patients with carcinoma. Extratumoral microfollicular tissue was found in the thyroid of patients above 50 years old mostly, and more rarely in young ones. The fragmented DNA concentration in microfollicular tissue was lower by a factor of 3.5 and intensity of stimulated internucleosomal DNA fragmentation was also decreased. Activity both of cathepsin B in lisosomes and caspase-3 in lysates of such tissue was also decreased. Conclusions: The decrease of intensity of spontaneous apoptosis and the absence of its modulation/induction following proapoptotic factors in extratumoral microfollicular thyroid tissue may be considered as a respond of the thyroid gland tissue to an existence of carcinoma. |
| format |
Article |
| author |
Myshunina, T.M. Kalinichenko, O.V. Tronko, N.D. |
| author_facet |
Myshunina, T.M. Kalinichenko, O.V. Tronko, N.D. |
| author_sort |
Myshunina, T.M. |
| title |
Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue |
| title_short |
Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue |
| title_full |
Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue |
| title_fullStr |
Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue |
| title_full_unstemmed |
Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue |
| title_sort |
abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2012 |
| topic_facet |
Original contributions |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/138690 |
| citation_txt |
Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue / T.M. Myshunina, O.V. Kalinichenko, N.D. Tronko // Experimental Oncology. — 2012. — Т. 34, № 2. — С. 112-115. — Бібліогр.: 21 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
AT myshuninatm abnormalitiesofapoptosisofthethyroidglandcellsfromextratumoralmicrofolliculartissue AT kalinichenkoov abnormalitiesofapoptosisofthethyroidglandcellsfromextratumoralmicrofolliculartissue AT tronkond abnormalitiesofapoptosisofthethyroidglandcellsfromextratumoralmicrofolliculartissue |
| first_indexed |
2025-07-10T06:22:24Z |
| last_indexed |
2025-07-10T06:22:24Z |
| _version_ |
1837239947514347520 |
| fulltext |
112 Experimental Oncology 34, 112–115, 2012 (June)
ABNORMALITIES OF APOPTOSIS OF THE THYROID GLAND CELLS
FROM EXTRATUMORAL MICROFOLLICULAR TISSUE
T.M. Myshunina, O.V. Kalinichenko, N.D. Tronko
State Institution “V.P. Komisarenko Institute of Endocrinology and Metabolism of National Academy
Medical Science”, Kyiv 04114, Ukraine
Aim: The aim of this study was to evaluate the frequency of existence of thyroid extratumoral normo- and microfollicular tissue in patients
with thyroid carcinoma and peculiarities of apoptosis in mentioned tissue. Materials and Methods: Using samples of normo- and micro-
follicular thyroid tissue it was determined the content of fragmented DNA and intensity of stimulated internucleosomal DNA fragmenta-
tion; activities of caspase-3 and cysteine lysosomal cathepsins. Results: It was found that normofollicular tissue is observed more often
in patients with nodal euthyroid goiter but microfollicular tissue is more common for patients with carcinoma. Extratumoral microfol-
licular tissue was found in the thyroid of patients above 50 years old mostly, and more rarely in young ones. The fragmented DNA con-
centration in microfollicular tissue was lower by a factor of 3.5 and intensity of stimulated internucleosomal DNA fragmentation was also
decreased. Activity both of cathepsin B in lisosomes and caspase-3 in lysates of such tissue was also decreased. Conclusions: The decrease
of intensity of spontaneous apoptosis and the absence of its modulation/induction following proapoptotic factors in extratumoral micro-
follicular thyroid tissue may be considered as a respond of the thyroid gland tissue to an existence of carcinoma.
Key Words: thyroid extratumoral microfollicular tissue, internucleosomal DNA fragmentation, caspase-3, lysosomal cathepsins, apoptosis.
A century ago, it was stated by Isaac Levin that tumor
development and growth depend not only on host’s im-
mune status, but also on local interactions between
tumor and host [1]. However, we have meanwhile only
a few information concerning metabolic changes in cells
of tumor-bearing organs. During last 50–60 years re-
searchers’ attention was mostly focused on the study
of tumor cell microenvironment that differs from normal
cell microenvironment without any doubt [2].
As an example of cell metabolism damage in any
organ-bearing tumor may be the data concerning lac-
tate formation in tumor cells due to aerobic glycolysis
activation that leads to the pH decrease not only in tu-
mor cells, but also in areas being outside of the tumor
by means of proton “leakage” [3]. Moreover more high
lactate dehydrogenase activity is registered in extratu-
mor tissue also [4]. Such acid microenvironment is toxic
for normal cells that leads to their destruction [3] and
well correlated with metastasis.Very often the availability
of the tumor is accompanied by various metabolic dis-
turbances even in areas being far from tumor setting [5].
An increased quantity of aberrant thyrocytes is de-
scribed for tumor-affected thyroid gland (TG) in the
all non-affected areas [6]; there are data concerning
both increased expression of some molecules taking
part in proliferation and apoptosis regulation [7] and
increased concentration of fragmented DNA in com-
pare to such in extranodular tissue in patients with
euthyroid goiter [8].
The TG activity is known may be changed in physio-
logical conditions or due to the thyroid pathology; the
follicular structure of TG tissue may also be changed:
the gland may have normo-, micro-, macro-, or hetero-
follicular structure. During the study of disturbances
in mechanisms of apoptosis in patients with benign
as well as malignant TG neoplasms we have observed
the existence of extratumoral tissue without histological
features of pathology in patients with carcinoma that
had more often microfollicular structure compare to pa-
tients with goiter. The aim of this work was to analyse
frequency of existence of thyroid extratumoral normo-
and microfollicular tissue of patients with thyroid car-
cinomas and peculiarities of apoptosis of such tissues.
MATERIALS AND METHODS
Tissue. It have been investigated extratumoral/
extranodular tissues (258 samples) obtained during
total or hemithyroidectomy to determine frequency
of existence of thyroid tissue of different follicular
structures without pathological changes when car-
cinoma or benign node (e.g., euthyroid goiter) are
present. All patients were informed about investigation
for scientific purposes. This study has been apptoved
by Institute’s Committee on Biological & Medical Ethics.
All tissue samples for biochemical investigation
(90 samples) had no macro- or microscopical fea-
tures of pathological changes. Depending on the type
of follicular structure all samples were divided into
three groups — tissue of normofollicular structure
(42 samples, 1st group), tissue of micro- or micro-
normofollicular structure (30 samples, 2nd group), and
tissue of macro- or macro-normofollicular structure
(18 samples, 3rd group).
Fragmented DNA assay. Concentration of low
molecular weight DNA fragments was analysed in su-
pernatant fluid following cell lysis during tissue homog-
enization in в 10 mM Na-ЕDTA-Tris-HCl buffer, рН 7.8,
containing 0.5% Triton Х-100 and centrifugation
(5.000 rpm, 15 min). The pellet containing high mo-
lecular weight DNA inside chromatin was suspended
in the initial buffer (but without Triton Х-100). Quan-
titative DNA content in both fractions was stimated
Received: February 15, 2012.
*Correspondence: E-mail: mishunina@list.ru
Abbreviation used: TG — thyroid gland.
Exp Oncol 2012
34, 2, 112–115
Experimental Oncology 34, 112–115, 2012 (June) 113
by means of a diphenylamine reaction [9] and given
as 1 μg/1 g of tissue. The part of fragmented DNA was
calculated as percent to the total amount.
The intensity of stimulated internucleosomal
DNA fragmentation assay. To determine the intensity
of stimulated internucleosomal DNA fragmentation, tis-
sue slices (50–100 mg) were previously incubated dur-
ing 4 h at 37 0С in Krebs-Ringer PBS (1 ml, pH 6.0) that
permited to increase the quantity of cells on the onset
of terminal step of apoptosis under hypoxia. The next
steps of DNA isolation and purification were the same
as discribed earlier [10]. DNA electrophoresis (1 h,
90 mV) was carried out in 1.7% agarose gel (“Sigma”,
USA, or “Lachema”, Czech Republic) using 10 mМ Tris-
EDTA buffer, рН 7.0–7.2 containing ethidium bromide
(1 μg/ml). The markers as large as 100–1000 base pairs
(bps) that have been used corresponded to DNA frag-
ments. Quantitative analysis of low molecular weight
DNA fragments (200, 400, 600, and 800 bps) was car-
ried out using the program “Photo Capt Mw”. Relative
nucleosome contents were calculated from total area
of all DNA peaks taken as 100%.
Caspase-3 and cysteine lysosomal cathepsin
activity assay. Obtaining of tissue lysates and deter-
mination of caspase-3 activity (acetyl-Asp-Glu-Val-
Asp-p-nitroanilide being as a substrate) were provided
using kit for estimation of caspase activity (“CASP-3”,
Sigma, USA). Activity of cysteine lysosomal cathepsins
was estimated both in cytosol and lysosomes accor-
ding to [11] using synthetic substrates N2-benzoyl-
DL-arginine-4-nitroanilide (for the cathepsin В) and L-
leucine-4-nitroanilide (for the cathepsin Н) or azocasein
being previously denatured by 6 M urea solution (for the
cathepsin L). Activity of caspase-3, cathepsin В, and
cathepsin Н was given as μmoles of p-nitroaniline that
are cleaved from the substrate during 1 h of incuba-
tion per 1 mg of protein. Activity of cathepsin L is given
in conventional units of low molecular weight peptide
(non-precipitated by trichloracetic acid) following
1 h of incubation per 1 mg of protein.
The allocation of lysosomal and cytosol frac-
tions, and protein concentration assay. Lysosomal
and cytosol fractions were isolated by means of dif-
ferential centrifugation and protein content was esti-
mated according to Lowry modification [12].
Statistical analysis. Data were given as mean ±
standard error (S.E.) and analyzed statistically using
a one-tail analysis of the Student’s t-test and χ2-test
for comparison between groups. Differences were
considered significant at p < 0.05.
RESULTS
Among 258 extranodal TG samples studied, dif-
ferent features of pathological changes (lymphoid
infiltration, invasion of tumor cells, chronic thyroiditis,
autoimmune thyroiditis, sclerotic changes of stroma
tissue, marked hyperplasia, cysts) were detected
in 168 ones (65,1%). Pathological events in extranodal
tissue were found mostly in patients with carcinomas
(131 samples/191; 68.5%) comparing to patients with
euthyroid goiter (37 samples/67; 55.2% p=0,0483,
χ2-test). Among the intact tissue samples with normal
follicular structure were mostly detected in nodular
euthyroid goiter (Table 1, p= 0.0071, χ2-test) and
tissue with microfollicular structure has been mainly
found at carcinomas (p = 0.00003). In such patho-
logical conditions extratumoral/extranodular tissue
of macrofollicular structure was seen with the same
frequency (p = 0.576).
Table 1. Existence of an extratumoral/extranodular tissue of a different
follicular structure in a TG of patients with carcinoma and a nodular euthy-
roid goiter
Patients 1st group 2nd group 3rd group
patients with carcinoma (n=60) 26 (43.3%) 26 (43.3%) 8 (13.4%)
patients with nodular goiter (n=30) 20 (66.7%)* 3 (10.0%)* 7 (23.3%)
Note: * — significant difference in the 1st and the 2nd group of patients,
p<0.05, χ2-test; n — number of observations.
Such differences in distributions of normo- and
microfollicular tissue in TG affected with carcinoma
do not depend on patients’ sex (p=0.182, χ2-test),
on tumor metastatic potential (p = 0.281), tumor inva-
sive properties (p = 0.943), or tumor location — tumor-
containing lobe or contralateral one (p=0.111). At the
same time, it may be clearly detected the dependence
of existence of tissue of different follicular structure
on patients’ age (extratumor microfollicular tissue
is mostly present in TG of patients above 50 years and
significantly rarer in young patients). On the contrary,
it was found an opposite situation for distribution
of normofollicular tissue (Table 2). No dependence has
been noted in the distribution of extranodal tissue with
different follicular structures with age of patients hav-
ing euthyroid goiter. The differences in existence of tis-
sue with different follicular structures are statistically
significant in 30–50 years old patients as well as above
50 years (р=0.0295 and 0.0183, respectively, χ2-text).
Table 2. Existence of an extratumoral/extranodular tissue of a different
follicular structure in a TG of different age patients
Age Cancer Goiter
n 1st group 2nd group n 1st group 2nd group
Under 30 year 18 13 (72.2%) 5 (27.7%) 6 5 (83.3%) 1 (16.6%)
31–50 years 21 12 (57.1%) 9 (42.9%) 14 13 (92.9%) 1 (7.1%)
Up 50 year 13 1 (7.7%)*# 12 (92.3%)*# 3 2 (66.7%) 1 (33.3%)
Note: * — significantly different from group under 30 year; # — from group
31–50 years, p<0.05, χ2-test, n — number of observations.
Concentration of fragmented DNA in tissue of mi-
crofollicular structure was by a 3.5 fold lower, its part
in total DNA content was by a 2.4 fold also lower com-
paring to tissue of normofollicular structure (Table 3).
It has been found that mononucleosome content
in samples of DNA from microfollicular tissue has been
decreased by 55%, total oligonucleosome content (the
nucleosome size was 200, 400, 600, and 800 bps) has
been by 23% lower than in samples of DNA isolated
from normofollicular tissues (Table 4).
Table 3. Concentration of the DNA in an extratumoral thyroid tissue
of a different follicular structure
Parameters 1st group (n=6) 2nd group (n=4)
High-molecular DNA (μg/g) 1.00 ± 0.09 0.94 ± 0.01
Low-molecular DNA (μg/g) 0.57 ± 0.07 0.16 ± 0.01*
Part of fragmented DNA (%) 35.81 ± 2.03 14.92 ± 0.51*
Note: * — significantly different from the 1st group, p < 0.05, Student’s t- test;
n — number of observations.
114 Experimental Oncology 34, 112–115, 2012 (June)
Table 4. Level of oligonucleosoms in DNA extracted from an extratumoral
thyroid tissue of a different follicular structure
Size of nucleosomes 1st group (n=6) 2nd group (n=4)
200 bp. (%) 8.77 ± 0.64 3.94 ± 0.94*
400 bp. (%) 5.45 ± 0.48 3.49 ± 1.34
600 bp. (%) 8.17 ± 0.41 7.19 ± 1.24
800 bp. (%) 6.63 ± 0.65 6.89 ± 1.12
Total amount (200–800) bps. (%) 29.0 ± 0.39 22.3 ± 2.87*
Note: * — significantly different from the 1st group, p < 0.05, Student’s t- test,
n — number of observations.
Caspase-3 activity in lysates of microfollicu-
lar tissue was significantly lower than in tissue
of normofollicular structure (0.038±0.022 versus
0.156±0.028 μmoles p-nitroaniline/h/mg of protein,
respectively, p<0.05). Activity both of cathepsins Н and
L in lysosomal and cytosol fractions isolated from nor-
mo- or microfollicular tissues do not differ significantly
(data not shown). Cathepsin B activity was drastically
lower in lysosomal fraction isolated from microfol-
licular tissue (22.45±3.29 versus 11.1±2.89 μmoles
p-nitroaniline/h/mg of protein, respectively, p<0.05).
DISCUSSION
While studying both intensity and mechanisms
of regulation of apoptosis under thyroid pathology
it was detected the prevalence of extratumor micro-
follicular tissue in presence of carcinoma that was
confirmed statistically. The extratumor microfollicular
tissue was seen more often (in a factor of 4.5) than
extranodular tissue of the similar structure in case
of euthyroid goiter. As it is known the microfollicular
structure of TG of healthy persons is characteristic
for children TG. Following aging, the TG becomes
more homogenous, follicles of average size are pre-
dominant. However, in healthy grown-up and older
persons it was observed the microfollicular changes
of TG architectonics more oft [13]. Such changes are
thought to be correlated with physiological TG changes
stipulated by aging. The increase of microfollicular
TG elements and the decrease of normofollicular ones
in extratumoral tissue of patients with carcinoma may
be related to their age. However, such dependence
is absent in patients with euthyroid goiter. That is why
the age cannot be the only cause of more oft occur-
rence of microfollicular extratumoral tissue in TG with
carcinoma. Other factors, such as patients’ sex, tumor
aggressiveness, location of tumor in TG are not crucial.
As it is known the decrease of follicle sizes takes
place due to the factors stimulating the TG func-
tion. It is shown that level of serum thyrotropin (TSH)
in patients with carcinoma may be increased, being,
however, not significantly higher than the upper normal
level [14–16]. Some authors think that such increase
is observed only in patients with papillary but not with
follicular carcinomas [15]. Other investigators have
found the increase of serum TSH concentration in pa-
tients with papillary, follicular, and undifferentiated car-
cinomas [16]. Consequently, despite of the absence
of clinical symptoms indicating the damage of thyroid
function in patients with carcinoma level of thyroid hor-
mones remains within normal ranges. It is impossible
to deny a possibility of definite stimulation TG by the
TSH (in a case of subclinical hypothyroidism) that
resulted in the decrease of follicle volume.
In extratumoral TG areas of microfollicular struc-
ture, it was not found any significant changes of lyso-
somal cysteine cathepsins H and L activity under such
stimulation. Taking into account their role in thyro-
globulin processing ( cathepsin L takes part in all steps,
but cathepsin H is active during complete protein
degradation [17]), it may be an indirect evidence that
there is absent significant quantitative shifts in synthe-
sis of thyroid hormones. At the same time, a drastical
droping of cathepsin B activity in lysosomes isolated
from microfollicular tissue may lead to small decrease
of content of thyroid hormones in blood: cathepsin
B takes part in this process at steps both of protein
solubilization and thyroxine formation [17]. It has been
earlier shown that thyroxine, but not triiodothyronine
content is something lower in blood of patients with
carcinoma [16]. Such decrease of circulating thyro-
xine (something lower than normal range) is proposed
to be as a cause of a little increase of TSH secretion
and, consequently, diminished follicle volume.
On the other hand, as it was ascertained earlier the
release of lysosomal proteases into the cytosol due
to permeability of lysosomal membrane is dominant
intracellular phenomenon but not as a part of the fol-
lowing later stages of cell death throughout the apop-
tosis. Releasing of proteases including cathepsins
from lysosomes causes the mitochondrial disfunction
followed by activation of caspases and cell death [18].
In this work, no cathepsin release into the cytosol from
microfollicular tissue has been registered. This fact
and our data obtained concerning the transmembrane
potential as well as intensity of swelling of mitochondria
isolated from such tissue [19] suggest that there are
absent activation of apoptosis in thyroid extratumoral
microfollicular tissue.
However, the spontaneous apoptosis in such tis-
sue may by inhibited. Such assumption is confirmed
by a drastic drop of caspase-3 (whose participa-
tion in apoptosis has been confirmed of numerous
researchers [20, 21]), as well as by low cathepsin
B activity (its role in regulation of apoptosis has been
already established [18]). The decrease of activity
of these proteases in microfollicular thyroid tissue
coincides also with the data concerning decreased
level of fragmented DNA (Table 3) and gradual delay
of swelling of mitochondria [19]. All mentioned above
and significant decline of mitochondrial transmem-
brane potential [19] may be an evidence of the change
of membrane permeability and decrease of proapoptic
molecule releasing from mitochondria.
In addition, some disturbances of mitochondrial
function in response to apoptosis modulating factors
in thyroid tissue have been shown earlier. It may be im-
portant for realization of cell death following proapop-
tosis signals. First of all, mitochondria in microfollicular
tissue becomes to be resistant to the affect of calcium
ions (pore opening inducers) and antioxidants that
can modulate this process. Intensity of mitochondria
Experimental Oncology 34, 112–115, 2012 (June) 115
swelling (the most adequate parameter permitting
to estimate the nonspecific mitochondrial membrane
permeability) has not been changed following calcium
ions, melatonin or α-tocopherol addition to the incuba-
tion medium that is inherent for normofollicular tissue.
The transmembrane potential is an integral parameter
describing energetic potential of mitochondria. Its de-
crease causes the release of proapoptic factors from
mitochondria into the cytosol. However calcium ions
promoted the increase of transmembrane potential
in some tissue samples [19]. The resistance of micro-
follicular cells to proapoptic signals is confirmed by our
data obtained: decrease of the intensity of terminal
stage of apoptosis — stimulated internucleosomic
DNA fragmentation.
It has been derermined the decrease of spontane-
ous apoptosis in extratumoral microfollicular thyroid
tissue and its resistance to proapoptotic factors may
be as a result of the tumor influence. It may be pro-
moted further tumor progression, as well as thyroid
tissue stimulation by means of TSH.
ACKNOWLEDGMENTS
We wish to thank Mrs. O.A. Ornatskaya for excellent
technical assistance and pathologist L.Yu. Zyrnadzhu
for detailed pathohistological examination of thyroid
tissue samples.
REFERENCES
1. Levin I. Changes in the tissue surrounding a growing
tumor and the significance of the “precancerous state”. J Exp
Med 1912; 16: 149–54.
2. Osinsky S, Vaupel P. Tumor Microphysiology. Naukova
Dumka, Kiev, 254 p, 2009 (in Russian).
3. Gatenby R, Gawlinski E, Gmitro A, et al. Acid-mediated
tumor invasion: a multidisciplinary study. Cancer Res 2006;
66: 5216–23.
4. Shatova O, Zinkovich I, Borzenko B. Glycolysis
peculiarities at a mammary gland cancer. Pathology 2008;
5: 133 (In Russian).
5. Bakurova E, Sokolovsky L, Nikonova A, et al. Peculiari-
ties of carbohydrates metabolism in processes of the regulated
and non-regulated proliferation in a stomach mucosa. Ukrai-
nian Biochem J 2002; 74: 22 (In Russian).
6. Belyakov I. Clinical and morphological features of nodu-
lar neoplasms of a thyroid gland and tactical approaches
to their operative treatment. Phd Thesis, Moscow, 2006, 26 p
(In Russian).
7. Paltsev M, Kogan Е, Tuntsova O, et al. Morphological
and molecular-genetic characteristic of a cancer, adenomas
and a surrounding tissue of a thyroid gland. Archiv pathol
1998; 60: 5–10 (In Russian).
8. Komarevtseva E. Detection of an apoptosis in nodal
goiter of a thyroid gland. Ukrainian Med. almanac 2006;
7: 168–70 (In Russian).
9. Barton K. Definition of concentration of DNA with
the help of diphenylamine. In: Methods in Nucleic Acids
Research. Mir, Moscow, 7–9, 1970 (In Russian).
10. Kostyuchenko N, Pushkarev V, Kashevarov G, et al.
Effects of N-acyletanolamines and various antimitotic agents
on apoptotic DNA fragmentation in conventionally normal and
tumor tissue of human adrenals. Exp Oncol 2005; 27: 215–9.
11. Barret A, Kirschke H, Cathepsins BH, and Cathep-
sins L. In: Methods in Еnzymology. L Loran (ed). Academic
Press, New-York, London, 1981; 80 (part C), 35–61.
12. Hartree T. Determination of protein: a modification
of Lowry method that gives a linear photometric response.
Anal Biochem 1972; 48: 422–7.
13. Blumenthal H, Perlstein I. The aging thyroid. A de-
scription of lesions and an analysis of their age and sex distri-
bution. J Am Geriatr Soc 1987; 35: 843–54.
14. Ivanov P, Kazeev K, Karetnikov U. The tiroliberin
test in diagnostics of a cancer of a thyroid gland. Problemy
Endocrinilogii 1984; 30: 34–8 (In Russian).
15. Komisarenko I, Slavnov V, Rubakov S, Luchitskij E.
Radioimmunoassay definition of the functional status of pitu-
itary-thyroid system in the thyroid gland cancer patients. Ukr
Radiol J 1997; 5: 22–4 (In Ukrainian).
16. Grinevich G, Bundyug G, Jugrinova L, et al. Pecu-
liarities of the endocrine status of the thyroid cancer patients
of different age. Oncologiya 2003; 5: 284–7 (In Russian).
17. Friedrichs B, Tepel C, Reinheckel T, et al. Thyroid
functions of mouse cathepsins B, K, and L. J Clin Invest 2003;
111: 1733–45.
18. Guicciardi M, Leis M, Gores G. Lysosomes in cell
death. Oncogene 2004; 23: 2881–90.
19. Myshunina T, Kalinichenko O. Peculiarities of permea-
bility of membranes of mitochondria of cells from areas of nor-
mal thyroid parenchyma with micro- and normofollicular
structure. Endocrinologiya 2008; 13: 35–44 (In Ukrainian).
20. Abu-Qare A, Abou-Donia M. Biomarkers of apopto-
sis: release of cytochrome c, activation of caspase-3, induction
of 8-hydroxy-2’-deoxyguanosine, increased 3-nitrotyrosine,
and alteration of p53 gene. J Toxicol Environ Health B Crit
Rev 2001; 4: 313–32.
21. Kuribayashi K, Mayes P, El-Deiry W. What are cas-
pases 3 and 7 doing upstream of the mitochondria? Cancer
Biol Ther 2006; 5: 763–5.
Copyright © Experimental Oncology, 2012
|