Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma
Aim of this article is to study the effects of Paclitaxel (Ptx), γ-irradiation (IR) and their combination on the growth of xenografted tumors derived from undifferentiated thyroid cancer cells.
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
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| Cite this: | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma / V.M. Pushkarev, D.V. Starenki, V.O. Saenko, M.D. Tronko, S. Yamashita // Experimental Oncology. — 2011. — Т. 33, № 1. — С. 24–27. — Біліогр.: 14 назв. — англ. |
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| author | Pushkarev, V.M. Starenki, D.V. Saenko, V.O. Tronko, M.D. Yamashita, S. |
| author_facet | Pushkarev, V.M. Starenki, D.V. Saenko, V.O. Tronko, M.D. Yamashita, S. |
| citation_txt | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma / V.M. Pushkarev, D.V. Starenki, V.O. Saenko, M.D. Tronko, S. Yamashita // Experimental Oncology. — 2011. — Т. 33, № 1. — С. 24–27. — Біліогр.: 14 назв. — англ. |
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| description | Aim of this article is to study the effects of Paclitaxel (Ptx), γ-irradiation (IR) and their combination on the growth of xenografted tumors derived from undifferentiated thyroid cancer cells.
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| first_indexed | 2025-12-07T15:44:33Z |
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24 Experimental Oncology 33, 24–27, 2011 (March)
EFFECTS OF PACLITAXEL AND COMBINATION OF THE DRUG
WITH RADIATION THERAPY IN AN IN VIVO MODEL OF ANAPLASTIC
THYROID CARCINOMA
V.M. Pushkarev1, D.V. Starenki2,*, V. O. Saenko3, M.D. Tronko1, S. Yamashita3
1 State Institution “V.P. Komisarenko Institute of Endocrinology & Metabolism”, AMS of Ukraine, Kyiv 04114,
Ukraine
2Health Sciences University of Hokkaido, Hokkaido 061-0293, Japan
3Nagasaki University, Nagasaki 852-8523, Japan
Aim: To study the effects of Paclitaxel (Ptx), γ-irradiation (IR) and their combination on the growth of xenografted tumors de-
rived from undifferentiated thyroid cancer cells. Materials and Methods: Experiments were performed in nude mice with tumors
developing from implanted undifferentiated thyroid carcinoma cells (FRO). Animals were treated with Ptx i.p. and exposed locally
to a single dose of 5 Gy of IR. Apoptosis in situ was detected using ApopTag Peroxidase Kit. Results: In the in vivo experiments,
IR significantly inhibited but did not abrogate tumor growth. Ptx effect was stronger, and the combination therapy with Ptx and
IR led to the decrease of tumor volume to 0-0.3% of the control (P < 0.01). The systemic administration of Ptx to the animals with
advanced tumors resulted in a profound tumor growth suppression and in apoptosis in tumor tissues in time-dependent manner.
Conclusion: The combination of Ptx and IR is a promising strategy for further preclinical and clinical trials aimed at the develop-
ment of new therapeutic approaches to the treatment of undifferentiated thyroid cancer.
Key Words: Paclitaxel, ionizing radiation, thyroid cancer.
Paclitaxel (Ptx) has been successfully used to treat
different types of human cancers [1]. Among those,
the possibility of its application in one of the most ag-
gressive human tumors, anaplastic thyroid cancer
(ATC), both as a monotherapy and in a combination
with other anti-tumor agents was demonstrated [2–5].
Since Ptx mainly affects the microtubules thus block-
ing cell division, usage of additional agents that would
damage DNA in tumor cells to cause genotoxic stress
is a plausible modality. One of the agents widely used
in combination with chemotherapy, is ionizing radiation
(IR). It is known that Ptx enhances tumor cell radio-
sensitivity [6–9], possibly due to cell cycle arrest
at G2/M phase in which the cells are considered
to be more sensitive to radiation [8, 10]. The data
obtained in our in vitro experiments showed that ion-
izing radiation significantly enhances proapoptotic
effect of Ptx in anaplastic cancer cells [11]. The focus
of this study was to elucidate the combined effect
of low doses of Ptx and radiation on the in vivo growth
of xenograft tumors developing from human undif-
ferentiated thyroid carcinoma cells implanted in im-
munocompromised mice.
MATERIALS AND METHODS
Nude mouse xenograft model. Animal experi-
ments described in this study were conducted in ac-
cordance with the principles and procedures outlined
in the Guide for the Care and Use of Laboratory Animals
of the Biomedical Research Center, Center for Frontier
Life Science (Nagasaki University, Nagasaki, Japan).
Human follicular undifferentiated carcinoma cells FRO
were initially provided by J. A. Fagin (University of Cin-
cinnati College of Medicine, Cincinnati, OH). Cells
were grown in RPMI 1640 supplemented with 5% fetal
bovine serum (FBS) and 1% penicillin/streptomycin (all
reagents from Invitrogen Life Technologies, Paisley,
UK) in a 5 % CO2 humidified atmosphere at 37 0C. FRO
cells (5 x 106 cells per animal) resuspended in RPMI
1640 were injected s.c. into both flanks of 8-week–old
female BALB/c nu/nu mice (Charles River Japan, To-
kyo), 9 animals per group. Tumor sizes were measured
each alternate day with calipers, and tumor volumes
were calculated according to the formula a2 x b x 0.4,
where a is the smallest tumor diameter and b is the
diameter perpendicular to a. Treatment with Ptx and
γ-irradiation started after the tumor size approached
100 mm3. Ptx (10 mg/kg/day) diluted in Cremophor
EL (Sigma, USA), ethanol and phosphate-buffered
saline (PBS, pH 7.4) (1:1:1 v/v/v/) was injected in-
traperitoneally (i.p.) daily for 7 days. Animals from
the control group received vehicle injections. For the
exposure to γ-irradiation, anesthetized animals were
fixed in stalls, and the tumors were locally exposed
to a single dose of 5 Gy (Pony PS-3100SB, radiation
source 137Cs, 0.662 MeV, 1 Gy/min) 4 days after Ptx
treatment had started. Tumor size was monitored for
four more weeks during which the body weight, feeding
behavior, and motor activity of each animal were used
as indicators of general health.
To determine the effect of low doses of Ptx on ad-
vanced tumors, animals were treated with i.p. Ptx
injections at a dose of 2.5 mg/kg/day daily for 7 days.
Tumor size was measured for 18 consecutive days.
Received: February 5, 2011.
*Correspondence: Fax: 0133-23-1782;
E-mail: starenki@hoku-iryo-u.ac.jp
Abbreviations used: ATC – anaplastic thyroid cancer; FBS– fetal
bovine serum; IR – ionizing radiation; PBS – phosphate-buffered
saline; Ptx – Paclitaxel.
Exp Oncol 2011
33, 1, 24–27
25 Experimental Oncology 33, 24–27, 2011 (March)
Histological estimation of apoptosis in the
tumors. Needle biopsies of tumor tissues were fixed
in 10% neutral-buffered formalin, and embedded
in paraffin. Apoptotic cells were detected in 5-µm sec-
tions with an ApopTag Peroxidase Kit (Intergen Co.,
Burlington, MA). Positively stained cells were counted
in four fields (x100) for each specimen, and the apop-
totic index was determined as the ratio of apoptotic
cell number to total cell number.
Statistical analysis. All data were expressed
as a mean ± SD. Differences between groups were ex-
amined for statistical significance using Kruskal—Wallis
test (nonparametric ANOVA), Mann — Whitney test and
one-way analysis of variance (ANOVA). P < 0.05 was
considered indicating statistical significance.
RESULTS AND DISCUSSION
Treatment of anaplastic thyroid cancer cells in vitro
with IR does not abrogate cell growth although moder-
ate doses (1–2 Gy) can rather effectively induce apop-
tosis and transient growth arrest [11, 12]. At first, this
effect of IR was confirmed in the in vivo experiments
(Fig. 1, a). FRO cells transplanted into mouse flanks
quickly formed tumors. Twenty days after implantation,
tumor size exceeded 1000% of the initial (100 mm3)
in the control animals. In line with the in vivo experi-
ments, IR significantly (P < 0.05) reduced tumor size
but did not prevent tumor growth. Treatment with Ptx
was more effective (P < 0.05 as compared to irradi-
ated group). From the 11th day after the beginning
of Ptx treatment, we observed a highly significant
reduction of tumor volume as compared to both the
control and to the initial volume. At 20–29 days tumor
volume was 0.8–1% of control. The combined treat-
ment with radiation and Ptx showed the enhanced
therapeutic effect (P < 0.05 as compared to the ir-
radiated animals). Seven days after the beginning
of treatment, tumor size was significantly decreased;
after 20–29 days it was 0.3% of the control (P < 0.01),
and in two animals the tumors completely regressed.
The combination of irradiation and Ptx was slightly
more effective than Ptx alone, but the difference was
insignificant. In in vitro experiments the additive effect
of both agents regarding caspase-3 activation and
PARP cleavage was observed [11].
Next, after finishing this experiment, the control
group was split into 1 control and 8 experimental
animals which were subjected to the treatment with
4 times lower doses of Ptx (2.5 mg/kg/day). Fig.
1 b shows that the tumor in the control animal con-
tinued to grow up to 1700 mm3 during further 18 days
whereas those in the animals receiving Ptx were evi-
dently decreased. This observation indicates an ef-
ficiency of rather low doses of Ptx even in advanced
tumors.
Examination of the extent of apoptosis in tumor tis-
sues in situ in the advanced tumor group showed that
after 11 days of treatment, Ptx effectively induced cell
death, the intensity of which increased over the next
18 days (Fig. 2).
0
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*
*
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or
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ol
um
e,
m
m
3
Control
Ptx
ІR
Ptx + ІR
0 7 11 16 20 29
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0 2 4 6 8 10 12 14 16 18 20
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*
*
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or
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m
m
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Day
Control
Ptx (2.5 mg/kg/day)
a
b
Fig. 1. Effect of IR and Ptx on the growth of tumor xenografts
in vivo. (a) Effect of IR and Ptx as of monoagents and in combina-
tion. Ptx was injected i.p. at a dose of 10 mg/kg/day for 7 days. For
IR-monotherapy and combination therapy, animals were exposed
to a single dose of 5 Gy, 4 days after Ptx treatment had started.
Data are presented as mean ± SD of 9 tumors. *P < 0.05 vs. con-
trol group (Mann — Whitney test); + P < 0.05 vs. irradiated group
(Kruskal — Wallis test). (b) Effect of low doses of Ptx on advanced
tumors. Animals were treated with Ptx i.p. injections at a dose
of 2.5 mg/kg/day for 7 days. Data are presented as mean ± SD,
n = 1 for the control group, n = 8 for the treatment group. *P <
0.05 vs. n = 9 at day 0 (Kruskal — Wallis test)
Despite the combined effect of Ptx and radiation
in various tumors have been under investigation for
a long time, the mechanism of radiosensitization is not
fully understood. It is known that exposure to IR acti-
vates a complex system of sensors, mediators, signal
transducers, and effectors. The latter two categories
include checkpoint kinases, CHK1 and CHK2 (trans-
ducers), and p53 and Cdc25A-C (effectors) [13].
Their activities undergo changes under the action
of both agents. The response of tumor cells to Ptx
does not necessarily require the intact p53 and related
signaling mechanisms [14 for rev.] as demonstrated,
in particular, by its high cytotoxicity in ARO cells (ini-
tially assumed to be anaplastic thyroid carcinoma cell
line but recently reclassified into colon carcinoma)
with inactive TP53 [2]. In this study, the potentiation
of cytotoxic effect of the combination therapy (Ptx
and radiation) in tumors originating from FRO cells
with wild-type p53 may perhaps be attributed in part
to the activation of p53-dependent signaling pathways.
In this case, the augmentation of apoptosis most likely
26 Experimental Oncology 33, 24–27, 2011 (March)
dominates over the mechanisms responsible for cell
recovery and senescence.
Our in vivo data clearly show that treatment of ana-
plastic thyroid carcinoma with Ptx induces apoptosis
in tumor tissue. Also, the use of moderate (1–5 Gy)
doses of IR may be helpful for enhancement of Ptx
effect on undifferentiated thyroid cancer xenografts,
which confirms the data obtained for other tumors
[6, 7]. In conclusion, the combination of Ptx with
IR seems to be a promising modality for further pre-
clinical and clinical trials for advanced thyroid cancer.
REFERENCES
1. Kingston DGI. The shape of things to come: Structural
and synthetic studies of taxol and related compounds.
Phytochemistry 2007; 68: 1844–54.
2. Pushkarev VM, Starenki DV, Saenko VA, et al.
Molecular mechanism of the effects of low concentrations of
taxol in anaplastic thyroid cancer cells. Endocrinology 2004;
145: 3143–52.
3. Xu G, Pan J, Martin C, et al. Angiogenesis inhibition in
the in vivo antineoplastic effect of manumycin and paclitaxel
against anaplastic thyroid carcinoma. J Clin Endocrinol Metab
2001; 86: 1769–77.
0
5
10
15
20
25
*
*
Ap
op
to
tic
c
el
ls
, %
Control Day 11 Day 20 Day 29
*
a
b
Control 11 days
20 days 29 days
Fig. 2. Apoptosis induced by Ptx in tumor xenografts. Animals with advanced tumors were treated with daily i.p. Ptx injections
at a dose of 2.5 mg/kg/day for 7 days. (a) Apoptotic cells in tumor tissue biopsies were detected at 11, 20, and 29 days, as described
in Materials and Methods section, (x100). (b) Apoptotic index in the tumors. Data are mean ± SD. *P < 0.01 (ANOVA)
27 Experimental Oncology 33, 24–27, 2011 (March)
4. Ain KB, Egorin MJ, DeSimone PA. Treatment of ana-
plastic thyroid carcinoma with paclitaxel: phase 2 trial using
ninety-six-hour infusion. Collaborative Anaplastic Thyroid
Cancer Health Intervention Trials (CATCHIT) Group. Thy-
roid 2000; 10: 587–94.
5. Yeung S-C J, She M, Yang H, et al. Combination chemo-
therapy including combretastatin A4 phosphate and paclitaxel
is effective against anaplastic thyroid cancer in a nude mouse
xenograft model. J Clin Endocrinol Metab 2007; 92: 2902–9.
6. Dey S, Spring PM, Arnold S, et al. Low-dose fractionated
radiation potentiates the effects of paclitaxel in wild-type and
mutant p53 head and neck tumor cell lines. Clin Cancer Res
2003; 9: 1557–65.
7. Spring PM, Arnold SM, Shajahan S, et al. Low dose
fractionated radiation potentiates the effects of taxotere in
nude mice xenografts of squamous cell carcinoma of head and
neck. Cell Cycle 2004; 3: 479–85.
8. Toiyama Y, Inoue Y, Hiro J, et al. The range of optimal
concentration and mechanisms of paclitaxel in radio-enhance-
ment in gastrointestinal cancer cell lines. Cancer Chemother
Pharmacol 2007; 59: 733–42.
9. Zhang AL, Russell PJ, Knittel T, et al. Paclitaxel en-
hanced radiation sensitization for the suppression of human
prostate cancer tumor growth via a p53 independent pathway.
Prostate 2007; 67:1630–40.
10. Safran H, Rathore R. Paclitaxel as a radiation sensitizer
for locally advanced pancreatic cancer. Crit Rev Oncol/
Hematol 2002; 43: 57–62.
11. Pushkarev VM, Starenki DV, Popadiuk ID, et al. The
effect of combined action of Taxol and ionizing radiation
on thyroid anaplastic cancer cells. Rep Ukr Acad Sci 2009;
№5: 209–12 (in Ukrainian).
12. Pushkarev VM, Starenki DV, Yamashita S, et al. The
enhancement of cytotoxic action of Taxol on thyroid anaplastic
cancer by γ-irradiation. Rep Ukr Acad Sci 2010; № 3: 190-3
(in Ukrainian).
13. Sancar A, Lindsey-Boltz LA, Unsal-Kacmaz K, Linn
S. Molecular mechanisms of mammalian DNA repair and the
DNA damage checkpoints. Annu Rev Biochem 2004; 73:39–85.
14. Tronko MD, Pushkarev VM, Popadiuk ID, et al. The
intracellular mechanisms mediating action of taxanes in tumor
cell. J АМS Ukraine 2010; 15, №3: 408–23 (in Ukainian).
Copyright © Experimental Oncology, 2011
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| id | nasplib_isofts_kiev_ua-123456789-32314 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-12-07T15:44:33Z |
| publishDate | 2011 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Pushkarev, V.M. Starenki, D.V. Saenko, V.O. Tronko, M.D. Yamashita, S. 2012-04-16T20:34:56Z 2012-04-16T20:34:56Z 2011 Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma / V.M. Pushkarev, D.V. Starenki, V.O. Saenko, M.D. Tronko, S. Yamashita // Experimental Oncology. — 2011. — Т. 33, № 1. — С. 24–27. — Біліогр.: 14 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/32314 Aim of this article is to study the effects of Paclitaxel (Ptx), γ-irradiation (IR) and their combination on the growth of xenografted tumors derived from undifferentiated thyroid cancer cells. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma Article published earlier |
| spellingShingle | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma Pushkarev, V.M. Starenki, D.V. Saenko, V.O. Tronko, M.D. Yamashita, S. Original contributions |
| title | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma |
| title_full | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma |
| title_fullStr | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma |
| title_full_unstemmed | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma |
| title_short | Effects of Paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma |
| title_sort | effects of paclitaxel and combination of the drug with radiation therapy in an in vivo model of anaplastic thyroid carcinoma |
| topic | Original contributions |
| topic_facet | Original contributions |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/32314 |
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