Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential
Aim. To study the effect of adaptor protein Ruk/CIN85 overexpression on the dynamics of migration and Matrigel invasion as well as transendothelial migration of murine 4T1 breast adenocarcinoma cells. Methods. Dynamics of 4T1 cells migration/invasion was monitored in real time using the xCELLigence...
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nasplib_isofts_kiev_ua-123456789-1542762025-02-23T17:18:01Z Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential Надекспресія адаптерного протеїну Ruk/CIN85 в клітинах аденокарциноми молочної залози миші лінії 4T1 супроводжується зростанням їх рухливості та інвазивного потенціалу Сверхэкспрессия адаптерного протеина Ruk/CIN85 в клетках аденокарциномы молочной железы мыши линии 4T1 сопровождается увеличением их подвижности и инвазивного потенциала Horak, I.R. Drobot, L.B. Borsig, L. Knopfova, L. Smarda, J. Structure and Function of Biopolymers Aim. To study the effect of adaptor protein Ruk/CIN85 overexpression on the dynamics of migration and Matrigel invasion as well as transendothelial migration of murine 4T1 breast adenocarcinoma cells. Methods. Dynamics of 4T1 cells migration/invasion was monitored in real time using the xCELLigence Real-Time Cell Analyzer (RTCA) DP Instrument equipped with a CIM-plate 16. Transendothelial migration (TEM) of 4T1 cells was performed through the layer of primary mouse lung endothelial cells seeded on gelatin-coated 24-well transwell inserts (8-μm pores).The two-tailed Student’s t-test for unequal variances was used for statistical analysis. Results. Ruk/CIN85-overexpression in 4T1 cells are indices a significantly increased motility, Matrigel invasiveness and migration through endothelial cells layer. Conclusions. The Ruk/CIN85 adaptor protein may play a potential role in the control of metastasis in vivo. Мета. Дослідити вплив надекспресії адаптерного протеїну Ruk/CIN85 на динаміку міграції й інвазії через Матригель, а також на ефективність трансендотеліальної міграції клітин аденокарциноми молочної залози миші лінії 4Т1. Методи. Динаміку міграції/інвазії клітин 4Т1 аналізували в режимі реального часу за допомогою приладу XCELLigence Real-Time Cell Analyzer (RTCA) DP Instrument, оснащеного імпедансним планшетом CIM-plate 16. Трансендотеліальну міграцію (ТЕМ) клітин 4Т1 здійснювали через шар первинних ендотеліоцитів легені миші, висіяних на мембрану (розмір пор 8 μм) камери Бойдена. Для статистичного аналізу використовували двовибірковий t-тест Ст’юдента для незалежних вибірок з нерівними дисперсіями. Результати. Встановлено, що надекспресія Ruk/CIN85 у клітинах лінії 4Т1 супроводжується значним зростанням рухливості, здатності до інвазії через Матригель та шар ендотеліальних клітин. Висновки. Отримані результати вказують на потенційну роль адаптерного протеїну Ruk/CIN85 у контролі метастазування in vivo. Цель. Исследовать влияние сверхэкспрессии адаптерного протеина Ruk/CIN85 на динамику миграции и инвазии через Матригель, а также на эффективность трансэндотелиальной миграции клеток аденокарциномы молочной железы мыши линии 4Т1. Методы. Динамику миграции/инвазии клеток 4Т1 анализировали в режиме реального времени с помощью прибора XCELLigence Real-Time Cell Analyzer (RTCA) DP Instrument, оснащенного импедансным планшетом CIM-plate 16. Трансэндотелиальную миграцию (ТЭМ) клеток 4Т1 осуществляли через слой первичных эндотелиоцитов легкого мыши, высеянных на мембрану (размер пор 8 μм) камеры Бойдена. Для статистического анализа использовали двухвыборочный t-тест Стьюдента для независимых выборок с неравными дисперсиями. Результаты. Установлено, что сверхэкспрессия адаптерного протеина Ruk/CIN85 в клетках линии 4Т1 сопровождается значительным ростом подвижности, способности к инвазии через Матригель и слой эндотелиальных клеток. Выводы. Полученные результаты указывают на потенциальную роль адаптерного протеина Ruk/CIN85 в контроле метастазирования in vivo. 2018 Article Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential / I.R. Horak, L.B. Drobot, L. Borsig, L. Knopfova, J. Smarda // Вiopolymers and Cell. — 2018. — Т. 34, № 4. — С. 284-291. — Бібліогр.: 33 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000981 https://nasplib.isofts.kiev.ua/handle/123456789/154276 577 en Інститут молекулярної біології і генетики НАН України Вiopolymers and Cell application/pdf |
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Structure and Function of Biopolymers Structure and Function of Biopolymers |
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Structure and Function of Biopolymers Structure and Function of Biopolymers Horak, I.R. Drobot, L.B. Borsig, L. Knopfova, L. Smarda, J. Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential Інститут молекулярної біології і генетики НАН України |
| description |
Aim. To study the effect of adaptor protein Ruk/CIN85 overexpression on the dynamics of migration and Matrigel invasion as well as transendothelial migration of murine 4T1 breast adenocarcinoma cells. Methods. Dynamics of 4T1 cells migration/invasion was monitored in real time using the xCELLigence Real-Time Cell Analyzer (RTCA) DP Instrument equipped with a CIM-plate 16. Transendothelial migration (TEM) of 4T1 cells was performed through the layer of primary mouse lung endothelial cells seeded on gelatin-coated 24-well transwell inserts (8-μm pores).The two-tailed Student’s t-test for unequal variances was used for statistical analysis. Results. Ruk/CIN85-overexpression in 4T1 cells are indices a significantly increased motility, Matrigel invasiveness and migration through endothelial cells layer. Conclusions. The Ruk/CIN85 adaptor protein may play a potential role in the control of metastasis in vivo. |
| format |
Article |
| author |
Horak, I.R. Drobot, L.B. Borsig, L. Knopfova, L. Smarda, J. |
| author_facet |
Horak, I.R. Drobot, L.B. Borsig, L. Knopfova, L. Smarda, J. |
| author_sort |
Horak, I.R. |
| title |
Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential |
| title_short |
Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential |
| title_full |
Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential |
| title_fullStr |
Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential |
| title_full_unstemmed |
Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential |
| title_sort |
overexpression of adaptor protein ruk/cin85 in mouse breast adenocarcinoma 4t1 cells induces an increased migration rate and invasion potential |
| publishDate |
2018 |
| topic_facet |
Structure and Function of Biopolymers |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/154276 |
| citation_txt |
Overexpression of adaptor protein Ruk/CIN85 in mouse breast adenocarcinoma 4T1 cells induces an increased migration rate and invasion potential / I.R. Horak, L.B. Drobot, L. Borsig, L. Knopfova, J. Smarda // Вiopolymers and Cell. — 2018. — Т. 34, № 4. — С. 284-291. — Бібліогр.: 33 назв. — англ. |
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Інститут молекулярної біології і генетики НАН України |
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284
I. R. Horak, L. B. Drobot, L. Borsig
© 2018 I. R. Horak et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Bio-
polymers and Cell. This is an Open Access article distributed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium,
provided the original work is properly cited
UDC 577
Overexpression of adaptor protein Ruk/CIN85 in mouse breast
adenocarcinoma 4T1 cells induces an increased migration rate
and invasion potential
I. R. Horak1, L. B. Drobot1, L. Borsig2, L. Knopfova3
1 Palladin Institute of Biochemistry, NAS of Ukraine
9, Leontovycha Str., Kyiv, Ukraine, 01601
2 University of Zurich
71, Rämistrasse, Zürich, Switzerland, CH-8006
3 Universitas Masarykiana Brunensis
617/9, Žerotínovo nám., Brno, Czech Republic, 601 77
iryna.horak@gmail.com
Aim. To study the effect of adaptor protein Ruk/CIN85 overexpression on the dynamics of
migration and Matrigel invasion as well as transendothelial migration of murine 4T1 breast
adenocarcinoma cells. Methods. Dynamics of 4T1 cells migration/invasion was monitored in
real time using the xCELLigence Real-Time Cell Analyzer (RTCA) DP Instrument equipped
with a CIM-plate 16. Transendothelial migration (TEM) of 4T1 cells was performed through
the layer of primary mouse lung endothelial cells seeded on gelatin-coated 24-well transwell
inserts (8-μm pores). The two-tailed Student’s t-test for unequal variances was used for statis-
tical analysis. Results. Ruk/CIN85-overexpression in 4T1 cells are indices a significantly
increased motility, Matrigel invasiveness and migration through endothelial cells layer.
Conclusions. The Ruk/CIN85 adaptor protein may play a potential role in the control of me-
tastasis in vivo.
K e y w o r d s: tumor cell migration, invasion, 4T1 cells, adaptor protein Ruk/CIN85.
Acquisition of increased motility rate and inva-
sion potential is a key prerequisite for meta-
static dissemination of malignantly trans-
formed cells followed by their maintenance in
the blood stream, arrest in the distant organ,
extravasation, homing and distant organ colo-
nization [1]. This extremely complex and mul-
tistage process is orchestrated by cellular plas-
ticity associated with adaptation of tumor cells
to dynamic changes in extracellular milieu,
which, in its turn, modulate the functional state
of signaling networks resulting in epigenetic
changes, induction or repression of specific
genes and miRNAs expression patterns [2, 3].
Structure and Function
of Biopolymers
ISSN 1993-6842 (on-line); ISSN 0233-7657 (print)
Biopolymers and Cell. 2018. Vol. 34. N 4. P 284–291
doi: http://dx.doi.org/10.7124/bc.000981
285
Overexpression Ruk/CIN85 in 4T1 cells induces an increased migration rate and invasion potential
Adaptor proteins are essential components of
signaling complexes. They not only assemble
the multimolecular complexes but also direct
and coordinate intracellular signaling in order
to fine-tune cellular behavior [4, 5, 6]. One of
such adaptor proteins is Ruk/CIN85, ubiqui-
tously expressed in a variety of normal and
cancer tissues in mammals (including human),
birds, reptiles, amphibians, bony fishes and
even insects (according to NCBI Gene tool).
Ruk/CIN85 consists of three SH3 domains,
proline- and serine-enriched regions, and
coiled-coil domain [7]. These binding domains
and motifs as well as sites for post-translation-
al modifications make Ruk/CIN85 a binding
partner for more than 200 proteins involved in
growth factors signaling, apoptosis, cell shape
signaling, cell adhesion, motility and invasion
[8, 9, 10]. Ruk/CIN85 was demonstrated to be
overexpressed in many human cancers, such
as gliomas [11], breast [12] and colon cancers
[13], head and neck squamous cell carcinomas
[14] in comparison to conditionally normal
surrounding tissues. Using tissue samples from
breast cancer patients, we previously demon-
strated that the highest levels of Ruk/CIN85
content was observed in the lymph node me-
tastases and intravascular tumor emboli [12]
that suggests its important role in the meta-
static process. In the present study we analyzed
the effect of Ruk/CIN85 overexpression in
highly invasive mouse breast adenocarcinoma
4T1 cells on the cellular features, required for
efficient metastasis: motility, invasiveness, and
transendothelial migration.
Materials and Methods
Cells and transfection. Murine breast adeno-
carcinoma 4T1 cells were cultured in RPMI-
1640 medium supplemented with 10 % fetal
calf serum, 2 mM L-glutamine, 100 U/ml
penicillin, 100 µg/ml streptomycin in a hu-
midified atmosphere containing 5 % CO2 at
37°C. Ruk/CIN85-overexpressing subline
RukUp and corresponding control subline were
obtained by Ca-phosphate transfection of 4T1
cells with pRc/CMV2-Rukl or empty vector,
respectively [12]. Transfected cells were se-
lected with 1 mg/ml geneticin (G418) followed
by subcloning.
Dynamics of cell migration/invasion.
Monitoring of 4T1 cells migration and inva-
sion in real time was performed using the
xCELLigence Real-Time Cell Analyzer
(RTCA) DP Instrument equipped with a CIM-
plate 16 (Roche, Indianapolis, IN) as described
previously [16]. For invasion experiments, the
membranes of CIM-plate 16 wells were coat-
ed with Matrigel and for migration experi-
ments they were left uncoated. 4T1 cells
(7,5×104) were added to the upper chamber of
the plate well in serum-free medium, and the
lower chamber was filled with 10 % FCS me-
dium. Also, as negative control we used wells
without FCS in the lower chamber (SF).
Migration/invasion was monitored every 10 or
15 min for several hours. For quantification,
the cell index at indicated time points was
averaged from at least three independent mea-
surements.
Transendothelial migration assay.
Transendothelial migration (TEM) assay was
performed as described previously [17].
Briefly, primary mouse lung endothelial cells
(2.5×104) were seeded on gelatin-coated
24-well transwell inserts (8-μm pores; BD
Biosciences) and were grown for two days to
reach the confluence. 4T1 cells of both anal-
286
I. R. Horak, L. B. Drobot, L. Borsig et al.
ysed sublines (2,5×104), pre-stained with
0,4 µM Calcein AM, were seeded into tran-
swell inserts with monocytes (1×105) purified
from bone marrow in 3 % FCS/RPMI. 10 %
FCS/RPMI was added into the bottom cham-
ber. As a negative control, we seeded mock-
transfected 4T1 cells alone (without mono-
cytes). After 24 hours transwell membranes
were fixed with 2 % PFA and nuclei were
stained with DAPI. Migrated 4T1 cells were
counted using fluorescent microscopy in at
least 20 consecutive viewfields.
Statistical analysis. All experiments were
performed at least in triplicates, and the data
were presented as Mean ± SD. For statistical
analysis we used two-tailed Student’s t-test for
unequal variances and difference between
groups was suggested to be significant at
p < 0,05.
Results and Discussion
In order to assess the effect of adaptor protein
Ruk/CIN85 on the motility and invasiveness
of 4T1 breast adenocarcinoma cells we used
4T1 cells with stable overexpression of full-
length Ruk/CIN85 isoform (RukUp subline)
and mock-transfected 4T1 cells as a control.
First, we analysed the dynamics of migra-
tion and Matrigel invasion of both RukUp and
control cells. For this purpose we used xCEL-
Ligence RTCA DP instrument that has inte-
grated Boyden chamber (CIM-Plate 16) and
allowed us to monitor motility and invasive-
ness in real time as a function of the impeded
electron flow through the membrane https://
www.aceabio.com/products/rtca-dp/ [18].
On the graph representing the dependence
of cell index on the time (Fig. 1), for both
analyzed sublines we observed a small peak
of cell index during first hour in migration and
invasion experiments, and this peak refers to
the attachment of cells to the membrane. Then,
approximately at 12th hour, RukUp and control
cells started to migrate/invade through the
membrane. It was demonstrated that Ruk/
CIN85-overexpressing 4T1 cells had signifi-
cantly higher cell index rate in comparison to
control cells in both migration and Matrigel
invasion experiments (Fig. 1 A, C). For migra-
tion rate, these differences were statistically
significant (p < 0,05) at 12, 16, 20 and 24 hrs
time points (Fig. 1 B) while, for Matrigel inva-
sion rate, – at 36, 48, 60 and 72 hrs time points
(Fig. 1 D).
Next, we compared the ability of RukUp
and control 4T1 cells to transmigrate through
the layer of endothelial cells in vitro using this
approach as a surrogate model of extravasation
process in vivo. After mouse lung endothelial
cells (ECs) seeded on the Boyden chamber
membrane reached the confluence, the inserts
were filled with tumor cells (2,5×104) togeth-
er with monocytes (1×105) allowing them to
migrate through the layer of ECs for 24 hrs.
As can be seen from Fig. 2, the efficiency of
TEM was about 18 times higher for Ruk/
CIN85-overexpressing 4T1 cells comparing to
the parental control cells.
Taken together, our findings evidence that
overexpression of adaptor protein Ruk/CIN85
in highly aggressive 4T1 cells led to further
increase of traits associated with breast cancer
progression and metastasis in vivo, especially
motility, Matrigel invasion and transendothe-
lial migration. These data were in agreement
with our previous results showing that Ruk/
CIN85-overexpressing human breast adenocar-
cinoma MCF-7 cells acquired increased motil-
287
Overexpression Ruk/CIN85 in 4T1 cells induces an increased migration rate and invasion potential
Fig. 1. Ruk/CIN85 affects the dynamics of migration and Matrigel invasion of 4T1 cells. A — Real-time migration of
4T1 RukUp and control cells; SF — negative control with serum-free medium in lower chamber of CIM-Plate. B —
Cell migration index of 4T1 RukUp and control cells at time points 12, 16, 20 and 24 hours. * — p < 0,05 in com-
parison to control. C — Real-time Matrigel invasion of 4T1 RukUp and control cells; SF — negative control. D — Cell
invasion index of 4T1 RukUp and control cells at time points 12, 24, 36, 48, 60 and 72 hours. * — p < 0,05 comparing
to control.
A
C
B
D
ity. Importantly, treatment of these cells with
the Src inhibitor PP2 and the PI3K inhibitor
LY294002 abolished the Ruk/CIN85-dependent
changes in cell motility [12]. The contribution
of Ruk/CIN85 to breast cancer malignancy was
further supported by the fact that reverse silenc-
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288
I. R. Horak, L. B. Drobot, L. Borsig et al.
ing of the adaptor protein in MCF-7 cells was
followed by inhibition of their motility [19].
Moreover, Cascio et al. showed that Ruk/
CIN85 promoted invasiveness and metastatic
potential of mouse B16 melanoma cells [20].
When we divided breast cancer cell lines from
a GSE44552 dataset (GEO) into TEM+ (cell
lines with high efficiency of transendothelial
migration) and TEM- (cell lines with low ef-
ficiency of transendothelial migration) (accord-
ing to [21]), it was found that every TEM+ cell
line expres ses higher levels of SH3KBP1
mRNA compared to TEM- cell lines (Fig. 3).
These data indicate that Ruk/CIN85 might be
involved in breast cancer metastatic cascade,
possibly by affecting interactions of tumor cells
with microvasculature.
An increasing number of reports have indi-
cated that in different types of human carcino-
mas acquisition of invasive and migratory
properties can be driven by an epithelial-to-
mesenchymal transition (EMT). This process
implies lack of apical-basal polarity, intercel-
lular contacts and adhesive properties, but
acquisition of front–rear polarity, changes in
the cell shape, organization of the cytoskele-
ton, ability to degrade the extracellular matrix
(ECM) [22, 23]. EMT can be induced by a
number of transcription factors, known as
“master regulators” of EMT, such as Twist,
Snail, Slug, Zeb 1/2 etc. These transcription
factors induce expression of mesenchymal
markers (vimentin, N-cadherin, fibronectin)
and repress expression of epithelial markers
(E-cadherin, claudins, β-catenin) [24, 25]. The
changes in expression levels and activity of
EMT-inducing transcription factors may occur
via signaling dependent on TGFβ receptor,
tyrosine kinase receptors (such as EGFR,
VEGFR, FGFR), Wnt, Notch, Hedgehog, in-
tegrins, inflammation or hypoxia [22, 26].
There are data suggesting a possible involve-
ment of adaptor protein Ruk/CIN85 in the
Fig. 2. Adaptor protein Ruk/CIN85 enhances transendo-
thelial migration of 4T1 cells. * — p < 0,05 comparing to
control cells.
0
4
8
12
16
20
W/O
Monocytes
Control RukUp
C
an
ce
r c
el
l n
um
be
r
*
Fig. 3. Expression of SH3KBP1 gene in TEM+ and
TEM- breast cancer cell lines.
289
Overexpression Ruk/CIN85 in 4T1 cells induces an increased migration rate and invasion potential
EMT regulation. First, Yakymovych and co-
authors [27] demonstrated that Ruk/CIN85
stimulates the presentation of TGFβ receptor
on the cell plasma membrane and thus regu-
lates TGFβ-dependent signaling. Also, Ruk/
CIN85 was shown to be involved in endocy-
tosis and sorting of receptor tyrosine kinases,
the well-known regulators of EMT process,
including EGFR [28, 29], Met [30] and
VEGFR-1[31]. Finally, Ruk/CIN85 is a bind-
ing partner of several proteins, involved in
migration, adhesion and cell shape regulation,
such as c-Cbl, endophilins, dynamins, FAK,
Src, and F-actin [9, 28, 32, 33]. All these fea-
tures of adaptor protein Ruk/CIN85 can con-
tribute to the control of tumor cell invasiveness
and metastatic potential.
In conclusion, to suppress the Ruk/CIN85-
induced malignant properties of tumor cells
revealed in our study, future investigations of
molecular mechanisms of its action are re-
quired.
Acknowledgements
This study was supported by SCOPES grant
No IZ73ZO from Swiss National Science
Foundation (SNSF) and by the State Fund for
Fundamental Research of Ukraine (pro-
ject F83).
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Надекспресія адаптерного протеїну Ruk/CIN85
в клітинах аденокарциноми молочної залози
миші лінії 4T1 супроводжується зростанням їх
рухливості та інвазивного потенціалу
І. Р. Горак, Л. Б. Дробот, Л. Борсіг, Л. Кнопфова
Мета. Дослідити вплив надекспресії адаптерного про-
теїну Ruk/CIN85 на динаміку міграції й інвазії через
Матригель, а також на ефективність трансендотеліаль-
ної міграції клітин аденокарциноми молочної залози
миші лінії 4Т1. Методи. Динаміку міграції/інвазії
клітин 4Т1 аналізували в режимі реального часу за
допомогою приладу XCELLigence Real-Time Cell
Analyzer (RTCA) DP Instrument, оснащеного імпеданс-
ним планшетом CIM-plate 16. Трансендотеліальну
міграцію (ТЕМ) клітин 4Т1 здійснювали через шар
первинних ендотеліоцитів легені миші, висіяних на
мембрану (розмір пор 8 μм) камери Бойдена. Для
статистичного аналізу використовували двовибірковий
t-тест Ст’юдента для незалежних вибірок з нерівними
дисперсіями. Результати. Встановлено, що надекспре-
сія Ruk/CIN85 у клітинах лінії 4Т1 супроводжується
значним зростанням рухливості, здатності до інвазії
через Матригель та шар ендотеліальних клітин.
Висновки. Отримані результати вказують на потен-
ційну роль адаптерного протеїну Ruk/CIN85 у контр-
олі метастазування in vivo.
К л юч ов і с л ов а: міграція пухлинних клітин, ін-
вазія, клітини 4T1, адаптерний протеїн Ruk/CIN85.
Сверхэкспрессия адаптерного белка Ruk/CIN85
в клетках аденокарциномы молочной железы
мыши линии 4T1 сопровождается увеличением
их подвижности и инвазивного потенциала
И. Р. Горак, Л. Б. Дробот, Л. Борсиг, Л. Кнопфова
Цель. Исследовать влияние сверхэкспрессии адаптер-
ного протеина Ruk/CIN85 на динамику миграции и
инвазии через Матригель, а также на эффективность
трансэндотелиальной миграции клеток аденокарцино-
мы молочной железы мыши линии 4Т1. Методы.
Динамику миграции/инвазии клеток 4Т1 анализиро-
вали в режиме реального времени с помощью прибо-
ра XCELLigence Real-Time Cell Analyzer (RTCA) DP
Instrument, оснащенного импедансным планшетом
CIM-plate 16. Трансэндотелиальную миграцию (ТЭМ)
клеток 4Т1 осуществляли через слой первичных эн-
дотелиоцитов легкого мыши, высеянных на мембрану
(размер пор 8 μм) камеры Бойдена. Для статистиче-
ского анализа использовали двухвыборочный t-тест
Стьюдента для независимых выборок с неравными
дисперсиями. Результаты. Установлено, что сверхэкс-
прессия адаптерного протеина Ruk/CIN85 в клетках
линии 4Т1 сопровождается значительным ростом под-
вижности, способности к инвазии через Матригель и
слой эндотелиальных клеток. Выводы. Полученные
результаты указывают на потенциальную роль адап-
терного протеина Ruk/CIN85 в контроле метастазиро-
вания in vivo.
К л юч е в ы е с л ов а: миграция опухолевых клеток,
инвазия, клетки 4T1, адаптерный протеин Ruk/CIN85.
Received 04.05.2018
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