Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues
Cytochrome CYP2E1 gene as well as c-fos and c-myc oncogenes expression have been studied in different tissues of Wistar rats kept at Chernobyl's exclusion zone. Reliable differences are revealed in above-mentioned genes expression on RNA level by comparing data obtained from experimental and co...
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| Zitieren: | Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues / V.I. Prima, L.G. Glushakova // Биополимеры и клетка. — 1998. — Т. 14, № 6. — С. 540-543. — Бібліогр.: 23 назв. — англ. |
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irk-123456789-1574862019-07-04T21:58:16Z Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues Prima, V.I. Glushakova, L.G. Структура и функции биополимеров Cytochrome CYP2E1 gene as well as c-fos and c-myc oncogenes expression have been studied in different tissues of Wistar rats kept at Chernobyl's exclusion zone. Reliable differences are revealed in above-mentioned genes expression on RNA level by comparing data obtained from experimental and control animal groups. Вивчали експресію гена цитохрому CYP2E1 а також онкогенів c-myc і c-fos у різних органах щурів лінії Wistar, що утримувалися в умовах Чорнобильської зони відчуження. При порівняні даних для дослідних і контрольних груп тварин виявлено вірогідні розбіжності в експресії вказаних генів на рівні РНК Изучали экспрессию гена цитохрома CYP2E1,a также онкогенов c-myc и c-fos в различных органах крыс линии Wistar, содержавшихся в условиях Чернобыльской зоны отчуждения При сравнении данных для опытных и контрольных групп животных выявлены достоверные различия в экспрессии, указанных генов на уровне РНК. 1998 Article Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues / V.I. Prima, L.G. Glushakova // Биополимеры и клетка. — 1998. — Т. 14, № 6. — С. 540-543. — Бібліогр.: 23 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.0004F7 http://dspace.nbuv.gov.ua/handle/123456789/157486 en Биополимеры и клетка Інститут молекулярної біології і генетики НАН України |
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Структура и функции биополимеров Структура и функции биополимеров Prima, V.I. Glushakova, L.G. Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues Биополимеры и клетка |
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Cytochrome CYP2E1 gene as well as c-fos and c-myc oncogenes expression have been studied in different tissues of Wistar rats kept at Chernobyl's exclusion zone. Reliable differences are revealed in above-mentioned genes expression on RNA level by comparing data obtained from experimental and control animal groups. |
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Article |
| author |
Prima, V.I. Glushakova, L.G. |
| author_facet |
Prima, V.I. Glushakova, L.G. |
| author_sort |
Prima, V.I. |
| title |
Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues |
| title_short |
Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues |
| title_full |
Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues |
| title_fullStr |
Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues |
| title_full_unstemmed |
Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues |
| title_sort |
radiation-induced peculiarities of cytochrome p-4502e1 and oncogenes mrna accumulation in different rat tissues |
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Інститут молекулярної біології і генетики НАН України |
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1998 |
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Структура и функции биополимеров |
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http://dspace.nbuv.gov.ua/handle/123456789/157486 |
| citation_txt |
Radiation-induced peculiarities of cytochrome P-4502E1 and oncogenes mRNA accumulation in different rat tissues / V.I. Prima, L.G. Glushakova // Биополимеры и клетка. — 1998. — Т. 14, № 6. — С. 540-543. — Бібліогр.: 23 назв. — англ. |
| series |
Биополимеры и клетка |
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AT primavi radiationinducedpeculiaritiesofcytochromep4502e1andoncogenesmrnaaccumulationindifferentrattissues AT glushakovalg radiationinducedpeculiaritiesofcytochromep4502e1andoncogenesmrnaaccumulationindifferentrattissues |
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2025-07-14T08:24:48Z |
| last_indexed |
2025-07-14T08:24:48Z |
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1837610030368555008 |
| fulltext |
ISSN 0233-7657 . Биополимеры и клетка. 1998. Т. 14. № 6
Radiation-induced peculiarities of cytochrome
P-450IIE1 and oncogenes mRNA accumulation
in different rat tissues
V. I. Prima, L. G. Glushakova
Institute of Molecular Biology and Genetics of National Academy of Sciences of Ukraine
150, Zabolotny str., Kyiv, 252143 , Ukraine
Cytochrome CYP2E1 gene as well as c-fos and c-myc oncogenes expression have been studied in different
tissues of Wistar rats kept at Chernobyts exclusion zone. Reliable differences are revealed in above-
mentioned genes expression on RNA level by comparing data obtained from experimental and control
animal groups.
The connection between the accumulation of c-fos
mRNA and Р-450ІІЕІ m R N A (B2mRNA x , CYP2E1
mRNA) during the prereplicative period of rat liver
cells division has been shown in previous work [1 ] .
Now we are interested in relat ionship between on
cogenes (c-fos and c-myc) and CYP2E1 gene exp
ression under radiat ion influence which takes place in
Chernobyl ' s exclusion zone. T h e interest is due to the
key role of both C-FOS, C-MYC and P-450IIE1
proteins in cell transformation [2, 31 . In this work the
attempt has been done to s tudy if any relationship
takes place between the expression of these genes on
RNA level after long action of low level radiat ion.
Materials and Methods . Animals. Two groups of
albino rats (Wistar) that have common ancestry were
bred at the animal care facilities of the Inst i tute of
Experimental Pathology, Oncology and Radiobiology
at Kyiv and at Chernobyl ' s exclusion zone. Male rats
were 4 months of age for all experiments reported
here. T h e whole-body dose of radioactivity absorbed
for this period by the ra ts living in Chernobyl through
inhalation and feeding amounted to 1.2 ± 0.3 cGy for
C s 1 3 7 . This is approximately 10 t imes higher than the
dose absorbed by the control ra t s .
Tissue acquisition. Different t issue samples were
used from the animals grown in Kyiv (control) and
Chernobyl ( i r radia ted) . T h e livers, bra ins , spleens
© V. I. P R I M A , L. G. G L U S H A K O V A , 1998
and testicles were removed, r insed in phosphate-
buffered saline ( N a 2 H P 0 4 , 1.5 g/1; NaCl, 8 g/1;
M g C l 2 6 H 2 0 , 0.1 g /1 ; K H 2 P 0 4 , 0.2 g /1 ; KC1, 0.2 g/ i ;
C a C l 2 - 2 H 2 0 , 0.132 g/1) and frozen in liquid nitrogen
These tissues were mainta ined at - 7 0 ° С All data
points involved pooled t issue samples of each kind
from 4—6 ra ts .
RNA isolation. Total RNA from the samples of
i r radiated and control animals was isolated using
guanidine thiocyanate by a procedure [4 j and quan
tified by measuring its absorbance at 260 nm; purity
was assessed by determining the 260 n.m/280 nm
ratio. After electrophoresis on denatur ing gels, RNA
integrity and loading equivalence were assessed by
ethidium bromide staining.
Northern blotting and hybridization, Samples of
total RNA were electroforetically size separated in 2Л
M formaldehyde —- 1 % agarose gels \5 I The RNA
was transferred to H y b o n d - N mem bra о i s u.Amer
sham», Great Britan) by a vacuum blotting method
and crosslinked to the membranes using ultravioSei
radiat ion. cDNA probes were randomly primed Ь
specific activity of at least 10 8 cpm [6 ]. Prehyir
ridization, hybridizat ion, and washing processes were
performed as described in [7] . Hybridized blots were
exposed to X-ray film at - 7 0 °С. Densitometer scans
of autoradiographs were performed on an LKB I Itro
scan XL Laser Densitometer . T h e valley-to-valley
method was used to calculate baselines, and da la
were normalized to the sum of signals. Ail blots were
540
R A D I A T I O N - I N D U C E D PECULIARITIES O F C Y T O C H R O M E Р-450ИЕІ
de-hybridized by incubation in H 2 0 at 90 °С for two
minutes twice. They were checked for total removal of
the probe by overnight exposure to X-ray film. Those
blots showing total removal of the initial probe were
then re-hybridized to a different cDNA clone.
cDNA clones. Probes were prepared from the
following clones: 1-kb PstI fragment of v-fos [ 8 ] ; the
Sall-Psfl fragment of pv-myc corresponding to myc
third-exon [9 J; 0.3-kb PstI fragment of CYP2E1
(cDNA of B2mRNA x [10]) .
Results and Discussion. T h e experiments repor
ted here were designed to determine whether the
expression of different cellular oncogenes as well as
cytochrome P-450 genes is al tered tn ra ts following
the long exposure to radioactive substances absorbed
by inhalation and feeding in Chernobyl ' s area . P r e
vious work from our laboratory has shown that the
dynamics of accumulation of c-fos mRNA is strongly
related with that of P-450 2E1 mRNA during the
prereplicative period of rat liver cells division [1 ] .
Experiments by other groups have shown modulation
of c-fos mRNA following exposure of cells in culture
to X-rays or other DNA-damaging agents [11—13] ,
possibly leading to cell t ransformation and neoplasia.
Fig. 1 it was shown by the microdensitometry results
for c-fos oncogene. These results demonstra te the
significant reduction of c-fos R N A level in almost all
the tissues of Chernobyl ' s animals relative to controls.
Fig. 2 represents the level of c-myc RNA in
different tissues of experimental and control ra ts .
These results demonst ra te c-myc RNA accumulation
in spleen and testicle cells. Expression of c-myc was
somewhat reduced in liver relative to controls, while
in brain it is essentially unaffected by the irradiation.
This suggests that the exposure of animals to radio
active substances absorbed by inhalation and feeding
causes different responses on the level of RNA
accumulation in different cell types, as well as selec
tive modulation of specific oncogenes. It should be
noted that the changes in expression observed here
are relatively small. This is in keeping with experi
ments from other groups demonstra t ing only mode
rate gene modulat ion following exposure to DNA-
damaging agents [11, 14—16] . Since increased exp
ression of some oncogenes (including c-myc) has been
shown to play a role in cellular transformation [1 j ,
our results also suggest that one mechanism of
i r radiat ion-mediated oncogenesis may involve the mo
dulation of oncogene expression in selected tissues.
On the o ther h a n d , it is of great importance the
decrease in activity of detoxification cell systems [17]
apparently caused by the damage of detoxifying
enzymes as well as their genes or mRNAs thu?
enhancing the negative action of radiation on cell
s tructures. For instance, it was recently demonstrated
that low level radiat ion (LLR) represses enzymatic
activity of superoxide dismutase [18] , glutathione
transferase and glutathione reductase [191 in human
tissues. Our prel iminary results also revealed some
repression of activity of a microsomal enzyme P-
Fig. 1. Analysis of c-fos RNA levels in different tissues of control and
irradiated animals. All micro denshomrtric data were normalized to
the sum of signals: / — Kyiv; 2 — Chernobyl
Fig. 2. Analysis of c-myc RNA levels in different tissues of contro;
and irradiated animals. All microdensitomrtric data were normali /x.
to the sum of signals: 1 — Kyiv; 2 — Chernobyl
541
PRIMA V. 1. G L U S H A K O V A L. G.
450IIE1 in rat liver under LLR influence (the data not
shown). At first, the last enzyme at tracted our
attention during the investigation of the early period
of rat liver regenerat ion caused by partial hepa-
tectomy (PH) . It was revealed that at the early hours
of regeneration the accumulation of messenger RNA
for protooncogene c-fos and gene of P-450IIE1 un
dergo sharp fluctuations but in reciprocal manner [1 ,
20 J. The gene CYP2E1 is a member of cytochrome
P-450 gene superfamily (CYP) encoding a group of
hemoproteins that mostly catalyze the oxidative meta
bolism of hydrophobic endogeneous compounds like
steroids, fatty acids, prostaglandins and exogenous
chemicals including drugs , carcinogens and environ
mental pollutants [21 ]. These substra tes can be
converted either to inert polar metabolites further
eliminated in a water-soluble form or to cytotoxic or
carcinogenic derivatives. Possible correlation of P -
450IIE genes with malignancy was noted in some
investigations 13, 22, ]. Therefore It is not surprising
that under the influence of radiat ion the expression of
gene CYP2E1 essentially changes, as Fig. 3 shows.
T h e RNA levels for this gene are very low in brain,
spleen, and testicles. Tha t is in accordance with the
previous ly revea led l iver-specif ic express ion of
CYP2E1 [23] . But there is obvious discrepancy
between the increasing of БІМА level and reduction of
enzymatic activity of this cytochrome in liver under
LLR. We suppose, it may be due to the radiat ion-
induced mutat ions of this gene, as well as other
damages on the way from DNA to protein.
Fig. 3. Analysis of CYP2EI RNA levels in different tissues of control
and irradiated animals. All microdensitomrtric data were normalized
to the sum of signals: 1 — Kyiv; 2 — Chernobyl
Asknowledgements. These studies were in part
s u p p o r t e d by N A T O L i n k a g e G r a n t n u m b e r .
H T E C H . L G 961147. We greatly appreciate helpful
discussions with Drs M. Bruschi. F. Guerlesquin, and
A. Doha.
B. L Прима, Л. Г. Глушакова
Индуцированные радиацией тканеспецифические особенности
содержания мРНК цитохрома P450I1E1 и онкогеиои у крысы
Резюме
Вивчали експресію гена цитохрому CYP2El> а також онкоген! в
c-myc і c-fos у різних органах щурів лінії Wistar, щ о утримува
лися в умовах Чорнобильської зони відчуження. При порівнянії:
даних для дослідних і контрольних груп тварин виявлено
вірогідні розбіжності в експресії вказаних генів на рівні РИК
В. И. Прима, Л. Г. Глуиіакова
Індуковані радіацією тканиноспецифічні особливості вмісту
мРНК цитохрому Р450НЕ1 та онкогеніп у щура
Резюме
Изучалаи экспрессию гена цитохрома CYP2EI,a также онкоге
нов c-myc и c-fos в различных органах крыс линии Wistw,
содержавшихся в условиях Чернобыльской зоны отчуждения
При сравнении данных для опытных и контрольных групп
животных выявлены достоверные различия в экспрессии, у ко
занных генов на уровне РНК.
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Received 26.01 .98
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