Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin
Aim: To compare ultrastructure, phenotypic profile and cell cycle progression ofMCF-7 human breast cancer cells and MCF7 sublines resistant to cisplatin (MCF-7/DDP) and doxorubicin (MCF-7/DOX). Methods: MTT-test, immunocytochemistry, flow cytometry, electron microscopy. Results: The development of...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2009
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| Цитувати: | Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin / N.Yu. Lukyanova, N.V. Rusetskya, N.A. Tregubova, V.F. Chekhun // Experimental Oncology. — 2009. — Т. 31, № 2. — С. 87–91. — Бібліогр.: 26 назв. — англ. |
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irk-123456789-1357012018-06-16T03:11:27Z Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin Lukyanova, N.Yu. Rusetskya, N.V. Tregubova, N.A. Chekhun, V.F. Original contributions Aim: To compare ultrastructure, phenotypic profile and cell cycle progression ofMCF-7 human breast cancer cells and MCF7 sublines resistant to cisplatin (MCF-7/DDP) and doxorubicin (MCF-7/DOX). Methods: MTT-test, immunocytochemistry, flow cytometry, electron microscopy. Results: The development of drug resistance to cisplatin and doxorubicin in MCF-7 cells upon the culturing of the initial cells with the raising concentrations of cytostatics was accompanied by the increase in cells adhesion, the increasing differentiation grade and the loss of steroid hormone receptors. Besides, it was shown that antiapoptotic mechanisms (decrease ofBcl-2 expression) and intracellular glutathione detoxifying system are involved in the process of cisplatin resistance development inMCF-7 cells. Atthe same time, P-glycoprotein overexpression in cells resistant to doxorubicin suggests MDR-dependent mechanism. Both doxorubicin- and cisplatin-resistant cells are characterized by the changes in the expression of several cell cycle regulators — Ki-67, cyclin D1, pRb and р21). Conclusion: The long-time culture of MCF-7 cells with cytostatic drugs results in the decreased cyclin D1, pRb, and Ki-67 expression and increased р21 expression with the increasing differentiation grade of the resistant cells. The underlying mechanisms of resistance to cisplatin and doxorubicin in MCF-7 cells may be different. 2009 Article Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin / N.Yu. Lukyanova, N.V. Rusetskya, N.A. Tregubova, V.F. Chekhun // Experimental Oncology. — 2009. — Т. 31, № 2. — С. 87–91. — Бібліогр.: 26 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/135701 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 Lukyanova, N.Yu. Rusetskya, N.V. Tregubova, N.A. Chekhun, V.F. Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin Experimental Oncology |
| description |
Aim: To compare ultrastructure, phenotypic profile and cell cycle progression ofMCF-7 human breast cancer cells and MCF7 sublines
resistant to cisplatin (MCF-7/DDP) and doxorubicin (MCF-7/DOX). Methods: MTT-test, immunocytochemistry, flow
cytometry, electron microscopy. Results: The development of drug resistance to cisplatin and doxorubicin in MCF-7 cells upon the
culturing of the initial cells with the raising concentrations of cytostatics was accompanied by the increase in cells adhesion, the
increasing differentiation grade and the loss of steroid hormone receptors. Besides, it was shown that antiapoptotic mechanisms
(decrease ofBcl-2 expression) and intracellular glutathione detoxifying system are involved in the process of cisplatin resistance development
inMCF-7 cells. Atthe same time, P-glycoprotein overexpression in cells resistant to doxorubicin suggests MDR-dependent
mechanism. Both doxorubicin- and cisplatin-resistant cells are characterized by the changes in the expression of several cell cycle
regulators — Ki-67, cyclin D1, pRb and р21). Conclusion: The long-time culture of MCF-7 cells with cytostatic drugs results
in the decreased cyclin D1, pRb, and Ki-67 expression and increased р21 expression with the increasing differentiation grade of the
resistant cells. The underlying mechanisms of resistance to cisplatin and doxorubicin in MCF-7 cells may be different. |
| format |
Article |
| author |
Lukyanova, N.Yu. Rusetskya, N.V. Tregubova, N.A. Chekhun, V.F. |
| author_facet |
Lukyanova, N.Yu. Rusetskya, N.V. Tregubova, N.A. Chekhun, V.F. |
| author_sort |
Lukyanova, N.Yu. |
| title |
Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin |
| title_short |
Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin |
| title_full |
Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin |
| title_fullStr |
Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin |
| title_full_unstemmed |
Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin |
| title_sort |
molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2009 |
| topic_facet |
Original contributions |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/135701 |
| citation_txt |
Molecular profile and cell cycle in mcf-7 cells resistant to cisplatin and doxorubicin / N.Yu. Lukyanova, N.V. Rusetskya, N.A. Tregubova, V.F. Chekhun // Experimental Oncology. — 2009. — Т. 31, № 2. — С. 87–91. — Бібліогр.: 26 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
AT lukyanovanyu molecularprofileandcellcycleinmcf7cellsresistanttocisplatinanddoxorubicin AT rusetskyanv molecularprofileandcellcycleinmcf7cellsresistanttocisplatinanddoxorubicin AT tregubovana molecularprofileandcellcycleinmcf7cellsresistanttocisplatinanddoxorubicin AT chekhunvf molecularprofileandcellcycleinmcf7cellsresistanttocisplatinanddoxorubicin |
| first_indexed |
2025-07-09T23:56:37Z |
| last_indexed |
2025-07-09T23:56:37Z |
| _version_ |
1837215671222534144 |
| fulltext |
Experimental Oncology 31, 87–91, 2009 (June) 87
Tumor drug resistance is one of the most impor-
tant problems in cancer treatment in general as well
as breast cancer treatment [1–3]. Many chemothera-
peutic schedules used in breast cancer treatment
include anthracyclines and platinum derivatives [2,
3]. Cisplatin belongs to the group of alkylating agents.
It binds to DNA bases causing crosslinks and breaks
in DNA strands interfering with DNA replication [4].
The antitumor effects of doxorubicin are associated
with DNA intercalation and degradation of the tubu-
lar apparatus caused by the active free radicals [5].
The mechanisms of drug resistance development
for these two drugs are different [4–7]. It is believed
that cisplatin resistance is caused by an expression
of proteins involved in glutathione-mediating detoxify-
ing pathways such as glutathione-S-transferase, glu-
tathione-reductase and glutathione itself [8, 9]. DOX
resistance, as believed, results from the overexpres-
sion of 170 kD Pgp glycopeptide, which is an energy-
dependent pump that effluxes xenobiotics away from
cells [10, 11]. It is also known that the development
of drug resistance phenotype could be accompanied
by changes in morphological structure, proliferative
potential and adhesion properties of cells as well as the
changes in expression of proteins involved in apoptosis
and cell cycle control [12–15]. Studies on mecha-
nisms of antitumor drug resistance development are
of paramount importance for further understanding
of fundamental processes in formation of drug resis-
tance phenotype in tumors with the aim of searching
the ways for overcoming such resistance.
The aim of the study was to compare the ultrastruc-
ture, phenotypic profile and cell cycle of MCF-7 human
breast cancer cells and sublines resistant to cytotoxic
effects of cisplatin (MCF-7/DDP) and doxorubicin
(MCF-7/DOX).
MATERIALS AND METHODS
Cell lines and drug treatment. For our studies
we used human breast cancer cell line MCF-7 and
its sublines resistant to cytotoxic effects of cisplatin
(MCF-7/DDP) and doxorubicin (MCF-7/DOX). The
cells of the initial MCF-7 line were cultivated in modi-
fied Dulbecco’s medium ISCOV (“Sigma”, Germany)
with addition of 10% of fetal calf serum (“Sangva”,
Ukraine) at the temperature of 37 °C and CO2 concen-
tration of 5%. Cells were reseeded twice a week at the
density 2–4 x 104 cells/cm2, when cell layer covered
about half of the flask surface.
The resistant variants MCF-7/DOX and MCF-7/DDP
were originated by growing initial MCF-7 cells with rai-
sing concentrations of cisplatin (from 0.01 to 6 µg/ml)
or doxorubicin (from 0.1 to 32 µg/ml), respectively.
Cisplatin and doxorubicin were added twice a week
after reseeding. Every two months, cell survival was
analyzed by MTT assay. IC50 values for MCF-7 and
MCF-7/DDP cells were 0.25 and 1 µg/ml of cisplatin,
respectively, and for MCF-7 and MCF-7/DOX cells —
0.5 and 8 µg/ml of doxorubicin, respectively. There-
fore, MCF-7/DDP were 4 times as much resistant
to the cytotoxic effect of cisplatin and MCF-7/DOX
cells were 16 times as much resistant to the cyto-
toxic effect doxorubicin as compared with the initial
MCF-7 cells.
MTT assay. Sensitivity to antitumor drugs (cis-
platin and doxorubicin) was measured every two
months using standard MTT-colorimetric test with
3-[4,5,dimethylthiasol-2-1]-2,5-diphenyltetrasolium
bromide (“Sigma”, Germany) [16].
Immunocytochemistry. Expression of surface and
intracellular antigens was studied immunocytochemi-
MOLECULAR PROFILE AND CELL CYCLE IN MCF-7 CELLS
RESISTANT TO CISPLATIN AND DOXORUBICIN
N.Yu. Lukyanova*, N.V. Rusetskya, N.A. Tregubova, V.F. Chekhun
R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology NAS of Ukraine, Kyiv
03022, Ukraine
Aim: To compare ultrastructure, phenotypic profile and cell cycle progression of MCF-7 human breast cancer cells and MCF7 sub-
lines resistant to cisplatin (MCF-7/DDP) and doxorubicin (MCF-7/DOX). Methods: MTT-test, immunocytochemistry, flow
cytometry, electron microscopy. Results: The development of drug resistance to cisplatin and doxorubicin in MCF-7 cells upon the
culturing of the initial cells with the raising concentrations of cytostatics was accompanied by the increase in cells adhesion, the
increasing differentiation grade and the loss of steroid hormone receptors. Besides, it was shown that antiapoptotic mechanisms
(decrease of Bcl-2 expression) and intracellular glutathione detoxifying system are involved in the process of cisplatin resistance deve-
lopment in MCF-7 cells. At the same time, P-glycoprotein overexpression in cells resistant to doxorubicin suggests MDR-dependent
mechanism. Both doxorubicin- and cisplatin-resistant cells are characterized by the changes in the expression of several cell cycle
regulators — Ki-67, cyclin D1, pRb and р21). Conclusion: The long-time culture of MCF-7 cells with cytostatic drugs results
in the decreased cyclin D1, pRb, and Ki-67 expression and increased р21 expression with the increasing differentiation grade of the
resistant cells. The underlying mechanisms of resistance to cisplatin and doxorubicin in MCF-7 cells may be different.
Key Words: drug resistance, cisplatin, doxorubicin, human breast cancer, MCF-7 cells, immunocytochemistry, cell cycle.
Received: May 3, 2009.
*Correspondence: Fax: 380442581656
E-mail: oncom@onconet.kiev.ua
Abbreviations used: DOX — doxorubicin.
Exp Oncol 2009
31, 2, 87–91
ORIgINAL CONTRIBUTIONS
88 Experimental Oncology 31, 87–91, 2009 (June)
cally using mouse monoclonal antibodies to P-glyco-
protein (Pgp), glutathione-S-transferase (GST), RE, PR,
p53, Bcl-2, E-cadherin, Ki-67, cyclin D1, pRb, c-myc,
p21 (“Dako Cytomation”, Denmark).
Flow cytometry. For cell cycle analysis
of MCF-7 cells and its sublines, resistant to cisplatin
(MCF-7/DDP) and doxorubicin (MCF-7/DOX), the cell
suspension (106 cells per 200 µl of saline) was washed,
2 ml of cold 70% ethanol was added, and suspension
was fixed on ice for at least 30 min. Specimens were
centrifuged for 5 min at 300 g with further super-
natant decantation. After resuspending the sample
in 400 µl of saline, 50 µl of RNAse (25 mg/ml) and
10 µl of propidium iodide (0.5 mg/ml) were added.
Samples were analyzed on PAS “Partec” flow cyto-
meter (Germany) using red filter [17].
Electron microscopy. The cells were fixed in 1.6%
glutharaldehyde solution in 0.1 M cacodylate buffer
(pH 7.3) for 1 h followed by washing in 0.1 M cacody-
late buffer solution for 16–18 h. To achieve the isotonic
state, sucrose (50 mg/ml) was added. Cell postfixa-
tion was done in 2% osmium tetraoxide with further
dehydratation in alcohols and embedding in araldite
as described elsewhere. The ultrathin sections pre-
pared on LKB-8800 ultratome and contrasted by uranil
acetate and lead citrate were examined in JEM-100B
electron microscope with 60 kV acceleration voltage
with further specimen photography [18].
Statistical analysis. Statistical analysis was done
using STATISTICA 6.0 software (StatSoft Inc., USA)
with acquisition of mean values and standard deviation
(SD). Student’s t-test was used to evaluate the signifi-
cance of the differences between groups. p > 0.05 was
considered as the significant difference.
RESULTS AND DISCUSSION
Morphological features of sensitive and re-
sistant to cisplatin and doxorubicin MCF-7 cells.
It is known, that the development of drug resistance
phenotype is accompanied by the changes in different
biological features of malignant cells, including mor-
phological ones. In contrast to spindle-shaped MCF-7
and MCF-7/DOX cells, MCF-7/DDP cells were more
spherical with high nucleus/cytoplasm ratio. In all three
lines, the nuclei were rounded with 2–4 nucleoli. Cells
resistant to doxorubicin and cisplatin were larger than
initial MCF-7 cells and were characterized by stronger
adhesion to the underlying surface.
Electron microscopy demonstrated more com-
plicated ultrastructural organization and increased
diffe rentiation grade upon formation of drug resis-
tance both to cisplatin and to doxorubicin (Fig. 1, a).
The number of the microtubules increased and the
fibers comprising the microfilaments of varying width
were evident. An active Golgi apparatus consisting
of 2–3 loci with many multivesicular bodies was also re-
vealed in the resistant cells (Fig. 1, b, c). The formation
of a lot of multivesicular bodies and their positioning
near the plasmatic membrane may be indicative of the
increased efflux of the foreign substances through cell
membrane, which decreases the amounts of cisplatin
and doxorubicin in cytoplasm of resistant cells and
their cytotoxic activity. In MCF-7/DDP cells, the system
of actin filaments located in the cortical layer of cyto-
plasm was activated (see Fig. 1, b), while in MCF-7/DOX
cells, the intermediate filaments were activated in the
central area of cytoplasm (see Fig. 1, c). Therefore, the
development of resistance to cisplatin and doxorubicin
in MCF-7 cells was accompanied by the significant
changes in ultrastructural organization suggestive
of the increasing differentiation grade.
a
b
c
Fig. 1. Electron microscopy of MCF-7 cells (a) and subclones
with drug resistance to cisplatin MCF-7/DDP (b) and doxorubicin
MCF-7/DOX (c)
Immunocy tochemical characterist ics
of MCF-7 cells, sensitive and resistant to cisplatin
and doxorubicin. Mdr1 gene amplification followed
by P-glycoprotein overexpression is one of the mecha-
nisms involved in the development of drug resistance.
An alternative mechanism is connected with GST
isoenzymes expression and increased metallothionein
expression [19–21]. Glutathione-S-transferases are
responsible for conjugating glutathione with different
xenobiotics. The detoxification of anticancer drugs
by metallothioneins is related to their linkage with
Experimental Oncology 31, 87–91, 2009 (June) 89
electrophilic antitumor drugs from cisplatin group,
because free metallothioneins are nucleophilic com-
pounds [21]. In our experiments, P-gp expression was
absent both in cisplatin-sensitive and resistant cells
(Table 1, Fig. 2, a) suggesting mdr1-independent way
of drug resistance. In contrast, in cytoplasm of most
doxorubicin-resistant cells (MCF-7/DOX), P-gp was
overexpressed (see Table 1; Fig. 2, c). Also we found
out the significant differences in GST expression
among the sublines studied. In cisplatin-resistant
cells, the increase in GST content (Fig. 2, b) along with
the decrease in the percentage of MT-positive cells
(see Table 1) was evident.
According to current knowledge, receptor status
of breast tumors is a prognostic factor that is con-
nected with their sensitivity to neoadjuvant therapy
and radiotherapy. We have reported earlier that the
drug resistance in MCF-7 human breast cancer cells
developed upon cell culture in raising cisplatin and
doxorubicin concentrations was accompanied by the
changes in hormone receptor expression [22]. Par-
ticularly, estrogen and progesterone receptors were
expressed in 20% and 40% respectively of the initial
cells, while in MCF-7/DDP and MCF-7/DOX cells these
receptors were absent (see Table 1). According to the
clinical experience, in a third of breast cancer patients
the worse prognosis is associated with the absence
of steroid hormone receptors.
It is known that cytotoxic activity of many antitumor
drugs is linked to their ability to induce apoptosis in tar-
get cells [23–25]. We have attempted to compare the
content of several apoptosis-related proteins in MCF-7
sublines with acquired drug resistance and in the ini-
tial MCF-7 cells. The acquisition of resistance to cis-
platin or doxorubicin was not associated with changes
of p53 expression, which was detected at a low level in all
cell lines studied (see Table 1). Antiapoptotic Bcl-2 pro-
tein was present in cytoplasm of almost all sensitive and
doxorubicin-resistant cells (Fig. 2, d, f) while the long-
time culture of MCF-7 cells in the presence of cisplatin
facilitated reduction of Bcl-2-positive cell percentage
from 80 to 10% (see Table 1; Fig. 2, e). Therefore, the
Table 1. Phenotypic features of breast cancer MCF-7 cells and sublines with induced resistance to cisplatin and doxorubicin
Cell line Cells, positive for studied proteins, %
RE PR Pgp GST MT p53 Bcl2 Ki-67 E-cad С-myc p21 рRb Cyclin D
MCF-7 202, 3 402, 3 02 03 702, 3 602, 3 602, 3 402, 3 902, 3 15 103 602, 3 402, 3
MCF-7/DOX/16 01 01 701, 3 03 101 501 151 101 101 10 20 401 201
MCF7/DDP/4 01 01 02 701, 2 152 501 101 201 201 10 201 301 201
In Table 1 and 2: 1significantly (p < 0.05) different from MCF-7; 2significantly (p < 0.05) different from MCF-7/DOX/16; 3significantly (p < 0.05) different from MCF-7/DDP/4.
a c
e
g
b
d f
h i
Fig. 2. Imunocytochemical features of MCF-7 cells with cisplatin and doxorubicin drug resistance phenotype: а — loss of Pgp
expression in sensitive MCF-7 cells; b — expression of GST in MCF-7/DDP cells; с — Pgp expression in MCF-7/DOX cells; d, е, f —
Bcl-2 expression in MCF-7, MCF-7/DDP and MCF-7/DOX, respectively; g, h — Ki-67 expression in sensitive MCF-7 cells and
MCF-7/DDP cells; i — р21 expression in MCF-7/DOX cells
90 Experimental Oncology 31, 87–91, 2009 (June)
development of cisplatin resistance in breast cancer
cells is associated with the lowered Bcl-2 expression.
These data agree with both experimental and clinical find-
ings demonstrating that high Bcl-2 expression predicts
chemosensitivity in breast and lung cancer.
We have also found decreased proliferative ac-
tivity of MCF-7 cells with resistance to cisplatin and
doxorubicin (Fig. 2, g, h) with three-fold reduction
of proliferative potential in MCF-7/DDP cells and two-
fold reduction in MCF-7/DOX cells (see Table 1). The
decrease in the proliferative potential may be corre-
lated with differentiation of the resistant cells. In fact,
both resistant variants (MCF-7/DOX and MCF-7/DDP)
were characterized by strong expression of E-cadherin
as compared to low expression of E-cadherin limited
to the tight intercellular contact only in the initial MCF-7
cells (see Table 1).
The decreased proliferative activity in cells acqui-
ring drug-resistant phenotype is believed to be as-
sociated with changes in the expression of some cell
cycle-controlling proteins [26]. In our study, all three
MCF-7 sublines expressed a low level of c-myc. The
development of cisplatin and doxorubicin resistance
in human breast cancer cells was accompanied by the
reduction of cyclin D1, pRb expression and the overex-
pression of p21 (Fig. 2, i). These findings correspond
with the available data of other authors [22, 26].
Therefore, the formation of drug resistance
to cisplatin and doxorubicin in human breast cancer
MCF-7 cells is characterized by changes in expression
of proteins involved in control of apoptosis, cell cycle,
proliferation, and adhesion.
Analysis of cell cycle in sensitive MCF-7 cells
and cells with resistance to cisplatin and doxoru-
bicin. The abnormal regulation of cell cycle is known
as one of the characteristic features of the malignant
cells. We have shown that the patterns of cell cycle
distribution in the initial MCF-7 cells are the same
as in cisplatin and doxorubicin-resistant sublines
(Table 2, Fig. 3). The mechanisms of cytotoxicity upon
cell exposure to cisplatin and doxorubicin are diffe rent.
It is believed that cisplatin is not a phase-specific drug
because it causes disorders in DNA transcription and
replication, which lead to cell cycle arrest and apoptosis.
In contrast to cisplatin, doxorubicin is a phase-specific
drug, affecting predominantly S and G2 phases of the
cell cycle. It was of interest to compare cell cycle tra-
verse in the initial and resistant MCF-7 cells upon cell
exposure to cisplatin and doxorubicin. We have shown
that in the initial MCF-7 cells, incubation with cisplatin
at a dose of IC10 for 24 h resulted in the significant de-
crease of S phase percentage (from 28.52% to 19.55%)
with G2/M arrest (from 16.37% to 33.4%). Meanwhile,
doxorubicin treatment resulted in the accumulation
of MCF-7 cells in G0/G1 phase with the G0/G1 cell per-
centage increasing from 55.11% to 76.8%. In contrast,
exposure to doxorubicin in MCF-7/DOX cells and ex-
posure to cisplatine in MCF-7/DDP cells had no effect
on cell cycle traverse (Fig. 4).
Table 2. Cell cycle distribution of MCF-7 cells and sublines with induced
resistance to cisplatin and doxorubicin
Cell line Percentage of cells in a phase of the cell cycle
G0/G1 S G2/M
MCF-7 55.11 28.52 16.37
MCF-7/DOX/16 61.05 27.25 11.73
MCF7/DDP/4 59.42 26.20 14.38
640
480
320
160
0
0 100 200 300 400 500
Co
un
ts
FL3-Pl
640
480
320
160
0
0 100 200 300 400 500
Co
un
ts
FL3-Pl
640
480
320
160
0
0 100 200 300 400 500
Co
un
ts
FL3-Pl
a
b
c
Fig. 3. Distribution of MCF-7, MCF-7/DDP, MCF-7/DOX cells
between cell cycle phases
11.73%
27.25%
61.05%
12.15%
11.05%
76.80%
59.42%
26.20%
14.38%
33.40%
49.30%
19.55%
0
10
20
30
40
50
60
70
80
90
G0/G1 S G2/M
MCF-7/Dox
MCF-7/S + Dox
MCF-7/CP
MCF-7/S + CP
%
Fig. 4. Changes in cell cycle as result of impact of studied
antitumor drugs
To sum up, the long-time culture of human breast
cancer MCF-7 cells in the presence of cisplatin or do-
xorubicin in vitro is accompanied by the pronounced
Experimental Oncology 31, 87–91, 2009 (June) 91
changes in molecular-biological properties of the cells
with both shared and drug-specific molecular mecha-
nisms of the formation of the resistant phenotype.
In both doxorubicin-resistant and cisplatin-resistant
cells, the steroid hormone receptors have been lost.
The development of cisplatin resistance involves the
antiapoptotic mechanisms (decreased Bcl-2 expres-
sion) and intracellular glutathione detoxifying system
while the development of doxorubicin resistance
seems to follow MDR-dependent mechanism sugges-
ted by P-glycoprotein overexpression. In cells resistant
to either cisplatin or doxorubicin, the adhesive proper-
ties are enhanced and the ultrastructural organization
is characterized of more complicated patterns implying
the increased differentiation grade. At the same time,
in resistant cells cyclin D1, pRb, and Ki-67 expres-
sion decreased while р21 expression increased. The
changed expression pattern suggests the decreased
proliferative potential of the cells with drug resistant
phenotype. Therefore, the resistant MCF-7 cells differ
from the initial cell line by the expression of proteins
associated with drug resistance as well as proteins
involved in control of apoptosis, proliferation and cell
adhesion. Our data may be advantageous for deve-
loping the novel schedules of anticancer treatment
accounting for the molecular-biological characteristics
of drug-resistant cells.
ACKNOWLEDgMENTS
This work was supported by Fundamental re-
searches state foundation (№ F28.4/047). We grate-
fully acknowledge Phd.M.P. Zavelevich (R.E. Kavetsky
Institute of Experimental Pathology, Oncology and
Radiobiology) for his helpful advice.
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