Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy
Photodynamic therapy leads to oxidative stress through the generation of free radicals. Oxidative stress causes damage to cellular macromolecules such as nucleic acids, proteins and lipids. Aim: To examine the hematoporphyrin derivative (HpD) — mediated photodynamic effect on cervical adenocarcinoma...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
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| Цитувати: | Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy / J. Saczko, W. Skrzypek, A. Chwilkovska, A. Choromanska, A. Pola, A. Gamian, J. Kulbaska // Experimental Oncology. — 2009. — Т. 31, № 4. — С. 195-199. — Бібліогр.: 39 назв. — англ. |
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irk-123456789-1382062018-06-19T03:03:49Z Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy Saczko, J. Skrzypek, W. Chwilkowska, A. Choromanska, A. Pola, A. Gamian, A. Kulbacka, J. Original contributions Photodynamic therapy leads to oxidative stress through the generation of free radicals. Oxidative stress causes damage to cellular macromolecules such as nucleic acids, proteins and lipids. Aim: To examine the hematoporphyrin derivative (HpD) — mediated photodynamic effect on cervical adenocarcinoma cell line HeLa. Methods: The HpD localization in HeLa cells was analyzed by confocal microscopy with epi-fluorescence system. Lipid peroxidation (LPO) was estimated by measurement of the concentration of malondialdehyde, protein degradation — by modified Ellman’s method, superoxide dysmutase (SOD) — using Ransod Kit. The expression of inducible nitric oxide synthase (iNOS) was detected by immunocytochemical staining. Results: The HpD was distributed all over the cytoplasm with preferential localization in the inner side of the plasma membrane and around the nuclear envelope. The process of photosensitizer distribution was time dependent. PDT-HpD increased the level of malonodialdehyde (MDA), SOD activity and the expression of iNOS in HeLa cells. However, PDT induced the decrease in the level of protein-associated thiol groups. Conclusions: Our study showed the important role of PDT-mediated oxidative stress in HeLa cells. HpD-PDT might be alternative and less invasive approach for treatment of patients with cervical cancer resistant for standard chemotherapy and radiotherapy. 2009 Article Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy / J. Saczko, W. Skrzypek, A. Chwilkovska, A. Choromanska, A. Pola, A. Gamian, J. Kulbaska // Experimental Oncology. — 2009. — Т. 31, № 4. — С. 195-199. — Бібліогр.: 39 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/138206 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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English |
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Original contributions Original contributions |
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Original contributions Original contributions Saczko, J. Skrzypek, W. Chwilkowska, A. Choromanska, A. Pola, A. Gamian, A. Kulbacka, J. Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy Experimental Oncology |
| description |
Photodynamic therapy leads to oxidative stress through the generation of free radicals. Oxidative stress causes damage to cellular macromolecules such as nucleic acids, proteins and lipids. Aim: To examine the hematoporphyrin derivative (HpD) — mediated photodynamic effect on cervical adenocarcinoma cell line HeLa. Methods: The HpD localization in HeLa cells was analyzed by confocal microscopy with epi-fluorescence system. Lipid peroxidation (LPO) was estimated by measurement of the concentration of malondialdehyde, protein degradation — by modified Ellman’s method, superoxide dysmutase (SOD) — using Ransod Kit. The expression of inducible nitric oxide synthase (iNOS) was detected by immunocytochemical staining. Results: The HpD was distributed all over the cytoplasm with preferential localization in the inner side of the plasma membrane and around the nuclear envelope. The process of photosensitizer distribution was time dependent. PDT-HpD increased the level of malonodialdehyde (MDA), SOD activity and the expression of iNOS in HeLa cells. However, PDT induced the decrease in the level of protein-associated thiol groups. Conclusions: Our study showed the important role of PDT-mediated oxidative stress in HeLa cells. HpD-PDT might be alternative and less invasive approach for treatment of patients with cervical cancer resistant for standard chemotherapy and radiotherapy. |
| format |
Article |
| author |
Saczko, J. Skrzypek, W. Chwilkowska, A. Choromanska, A. Pola, A. Gamian, A. Kulbacka, J. |
| author_facet |
Saczko, J. Skrzypek, W. Chwilkowska, A. Choromanska, A. Pola, A. Gamian, A. Kulbacka, J. |
| author_sort |
Saczko, J. |
| title |
Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy |
| title_short |
Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy |
| title_full |
Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy |
| title_fullStr |
Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy |
| title_full_unstemmed |
Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy |
| title_sort |
photo-oxidative action in cervix carcinoma cells induced by hpd — mediated photodynamic therapy |
| 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/123456789/138206 |
| citation_txt |
Photo-oxidative action in cervix carcinoma cells induced by HpD — mediated photodynamic therapy / J. Saczko, W. Skrzypek, A. Chwilkovska, A. Choromanska, A. Pola, A. Gamian, J. Kulbaska // Experimental Oncology. — 2009. — Т. 31, № 4. — С. 195-199. — Бібліогр.: 39 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
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| first_indexed |
2025-07-10T05:20:26Z |
| last_indexed |
2025-07-10T05:20:26Z |
| _version_ |
1837236044758515712 |
| fulltext |
Experimental Oncology 31, 195–199, 2009 (December) 195
Cervical carcinoma is the most frequent cause of
death from cancer in women from developing coun-
tries and areas where social changes and urbanization
may increase HPV infection among young genera-
tions [1]. Therefore, new diagnostic and therapeutic
techniques with significantly improved sensitivity and
specificity are required. Photodynamic therapy (PDT)
is still a developing method in the arsenal of anticancer
therapy, based on photosensitizer and the visible light.
This treatment leads to the generation of reactive oxy-
gen species (ROS), such as singlet molecular oxygen,
hydroxyl radicals, and/or superoxide anions [2]. The
use of PDT as cancer therapy is particularly advanta-
geous because of its intrinsic dual selectivity. The
photosensitizer localizes in the malignant tissue and
the light is also spatially focused on the lesion [3].
In PDT, photosensitizer is accumulated in tumor cells,
which is followed by irradiation with visible light. The oxi-
dative stress and ROS are factors which initiate the cell
death via apoptosis or necrosis in PDT [4, 5]. Although
PDT is designed to cause a cytotoxic reaction in tumor
tissue, a post-PDT response involving inflammatory,
innate, and adaptive immune reactions is envisioned
to assist in successful eradication of residual surviving
tumor cells [2, 6]. The reactions of free radicals with un-
saturated lipids and proteins in membranes may directly
cause alterations of membrane function. Mitochondrial
photosensitization is of particular interest because of the
key role performed by mitochondria in cell metabolism as
well as in the regulation of normal cell functions [7, 8].
Therefore, the intracellular accumulation of the
photosensitizers is one of the most important factors
to determine the efficacy of the PDT [9, 10]. Depen-
ding on their physicochemical properties and their
uptake mechanism, sensitizers can achieve different
intracellular concentrations and localize in different
subcellular compartments. Cell death mechanism
after PDT is determined by the preferential localization
of a sensitizer in target organelles [11, 12].
The effective antioxidant defence system requires
an increase in antioxidant enzyme activity, not only to
eliminate the initial radicals, but also the more toxic pro-
ducts of spontaneous free-radical reactions [13]. One of
the most important antioxidant enzymes is superoxide
dismutase (SOD), which catalyzes the dismutation of the
superoxide anion O2
– to O2 and H2O2. SOD can partially
prevent the photodestruction caused by PDT [14, 15].
Nitric oxide (NO) is generated by the oxidation of
arginine, a reaction catalyzed by the NO synthase (NOS)
enzyme. NO has become recognized as a major effector
molecule in diverse array of physiologic and pathologic
processes [16–19]. A major mechanism for cell injury
produced by NO in vivo is due to its diffusion-limited
reaction with superoxide to form peroxynitrite [20]. Re-
cently, NO production was demonstrated in drug-treated
photosensitized tumor cells [21]. NO plays an important
dualistic role in malignant cells after PDT. In tumors pro-
ducing high levels of NO, the PDT-induced reduction in
tumor blood flow, vascular occlusion and consequent
PHOTO-OXIDATIVE ACTION IN CERVIX CARCINOMA CELLS
INDUCED BY HPD — MEDIATED PHOTODYNAMIC THERAPY
J. Saczko1, *, W. Skrzypek2, A. Chwiłkowska1, A. Choromańska1, A. Poła4, A. Gamian1, 3, J. Kulbacka1
1Department of Medical Biochemistry, Silesian Piasts University of Medicine, 50-368 Wroclaw, Poland
2St. Hedwigs Hospital, 55-100 Trzebnica, Poland
3Department of Immunology of Infectious Diseases, Institute of Immunology and Experimental Therapy,
Polish Academy of Sciences, 53-114 Wroclaw, Poland
4Department of Biophysics, Silesian Piasts University of Medicine, 50-368 Wroclaw, Poland
Photodynamic therapy leads to oxidative stress through the generation of free radicals. Oxidative stress causes damage to cellular
macromolecules such as nucleic acids, proteins and lipids. Aim: To examine the hematoporphyrin derivative (HpD) — mediated pho-
todynamic effect on cervical adenocarcinoma cell line HeLa. Methods: The HpD localization in HeLa cells was analyzed by confocal
microscopy with epi-fluorescence system. Lipid peroxidation (LPO) was estimated by measurement of the concentration of malondi-
aldehyde, protein degradation — by modified Ellman’s method, superoxide dysmutase (SOD) — using Ransod Kit. The expression
of inducible nitric oxide synthase (iNOS) was detected by immunocytochemical staining. Results: The HpD was distributed all over
the cytoplasm with preferential localization in the inner side of the plasma membrane and around the nuclear envelope. The process of
photosensitizer distribution was time dependent. PDT-HpD increased the level of malonodialdehyde (MDA), SOD activity and the
expression of iNOS in HeLa cells. However, PDT induced the decrease in the level of protein-associated thiol groups. Conclusions:
Our study showed the important role of PDT-mediated oxidative stress in HeLa cells. HpD-PDT might be alternative and less invasive
approach for treatment of patients with cervical cancer resistant for standard chemotherapy and radiotherapy.
Key Words: cervical adenocarcinoma, photodynamic treatment, HpD, lipid peroxidation, thiol groups, nitric oxide synthase,
superoxide dismutase.
Received: June 1, 2009.
*Correspondence: Fax: +48 71 784 00 85;
E-mail: michal@bioch.am.wroc.pl
Abbreviations used: HpD — hematoporphyrin derivative; iNOS —
inducible nitric oxide synthase; LPO — lipid peroxidation; MDA —
malonodialdehyde; NO — nitric oxide; PDT — photodynamic therapy;
ROS — reactive oxygen species; SOD — superoxide dismutase.
Exp Oncol 2009
31, 4, 195–199
ORIGINAL CONTRIBUTIONS
196 Experimental Oncology 31, 195–199, 2009 (December)
ischemia may be diminished, while the inflammatory
reaction triggered by PDT may be suppressed. On the
other hand, elevated NO levels may maintain vessel dila-
tion during PDT light treatment, which can diminish the
decrease in tumor oxygenation and sustain in this way the
oxygen-dependent generation of phototoxic damage.
PDT induces inflammation, afterwards the tumor cells
might activate macrophages to produce iNOS, which
results in excessive NO release [17, 22, 23].
In the current study we applied PDT on in vitro
model of human cervical adenocarcinoma HeLa cells.
The aim of this study was to evaluate HpD-mediated
photodynamic effect on HeLa cells with the particular
focus on oxidative stress factors, such as LPO, the level
of –SH groups, SOD activity and immunocytochemical
determination of inducible NO synthase.
MATERIALS AND METHODS
Cell culture. HeLa cell line was purchased from
Institute of Immunology and Experimental Therapy,
Polish Academy of Sciences. The cell line was routinely
propagated in monolayer cultures in Dulbecco’s modified
Eagles’ medium (Sigma) supplemented with 10% fetal
bovine serum and glutamine with penicillin/streptomycin
(Sigma) in 25 cm2 Falcon flasks. The cells were maintained
in a humidified atmosphere at 37 °C and 5% CO2.
Photodynamic treatment. The cells were treated
with 30 μg/mL HpD (Hematoporphyrin «D», Porphy-
rin Products Inc., USA) in complete media, for 4 h in
the dark. Then cells were irradiated with a light dose
of 3 and 6 J/cm2 using lamp (10 mW/cm2) (OPTEL,
Opole, Poland) with polarized light and red filter
(632.8 nm). After irradiation the cells were incubated
in a humidified atmosphere at 37 °C and 5% CO2 for 0,
3 and 18 h. Control cells with HpD were incubated in
the same conditions.
The localization of HpD. Cells were grown on cover
glasses (24 x 24 mm, Thermo Scientific), and incubated
for 4 h with HpD at the concentration of 30 μg/mL. Af-
ter incubation, the cells were washed in PBS and then
fixed in 4% formalin buffer, washed 2 times in PBS and
water (Aqua Pro Injectione, Polpharma), then examined
by confocal microscopy with epi-fluorescence system
(Nikon Eclipse TE2000-E) using filters with an excitation
wavelength of 528–553 nm and emission wavelength of
578–633 nm.
Lipid peroxidation. The LPO was determined by
measurement of the final product of fatty acid per-
oxidation — MDA according to modified procedure
described by Kulbacka et al. [24]. The cells after pho-
todynamic treatment (as described above) in vitro were
trypsinized (0.25% Trypsin-EDTA), washed twice in
PBS, and re-suspended in 200 μl of PBS. The concen-
tration of MDA was quantified spectrophotometrically
( = 535 nm) using a set of MDA standards.
Proteins damage. The evaluation of protein deg-
radation is based on modified Ellman’s method. This
method uses reaction of DTNB acid with thiol groups
of proteins. The level of –SH groups was measured
spectrophotometrically on the basis of the absorbance
at the wavelength of 412 nm [25].
Determination of SOD activity. After photody-
namic treatment the cells were trypsinized and washed
twice in PBS. Then the cells were suspended in 50 mM
PBS, pH 7, with a mixture of protease inhibitors (Com-
plete Mini EDTA-free, Roche). The total intracellular
SOD activity was measured using a Ransod assay
(Randox Laboratories Ltd., Antrim, United Kingdom)
according to the manufacturer’s protocol.
Expression of iNOS. The cells were plated into 8-dip
glasses (Nunc). For immunocytochemical detection of
iNOS rabbit polyclonal anti-iNOS antibodies were used
(1:100, Santa Cruz, USA). Formalin-fixed (4%) cells were
immunostained by DAKO LSAB 2 kit. The % of stained
cells was determined by counting 100 cells in three
randomly selected fields, performed by two indepen-
dent investigators. Cells were considered as positive
if staining was observed in > 5% of cells. The intensity
of immunocytochemical staining was evaluated as fol-
lows: (–) negative, (+) weak, (++) moderate and (+++)
strong. Positive and negative controls were included.
Two independent experiments were performed for each
PDT combination.
Statistical analysis. Normality of continuous
variables was checked by the Shapiro-Wilk test. The
significance of the difference between mean values of
different groups of cells in comparison to control un-
treated cells was assessed by Student’s t-test, values
of p ≤ 0.05 were considered significant.
RESULTS
Localization of HpD. The intracellular distribution
of HpD monitored after 4 h of treatment is presented
on Fig. 1A and B. We could observe the dye diffused
throughout the cytoplasm. After 4 h the most intensive
signal was detected next to the inner side of the plasma
membrane (Fig. 1, a, b).
The evaluation of LPO. After PDT, the level of
LPO increased in cells that received the highest dose
of irradiation (6 J/cm2) at all pre-incubation periods.
Immediately post PDT and 3 h after it the concentra-
tion of MDA was on the same level (0.4–0.6 μM/L).
18 h after photodynamic treatment we have observed
a significant decrease of MDA level, which was below
the control (Fig. 2, a).
The evaluation of protein damage. The proteins
degradation was observed for all incubation time
points after PDT application, but mainly immediately
after PDT and 3 h after it. After 18 h of irradiation, we
observed double rise of the protein-associated –SH
groups concentration in comparison to control un-
treated cells (Fig. 2, b) (p < 0.05).
SOD activity. We observed that SOD activity
achieved the highest level (to about 20 U/mg of protein)
directly after irradiation and after 18 h post PDT. How-
ever 3 h post irradiation the SOD activity decreased to
14 U/mg of protein (Fig. 2 C). The highest value of the
enzyme activity was observed for the cells that received
the highest dose of irradiation (6 J/cm2). The values
Experimental Oncology 31, 195–199, 2009 (December) 197
of enzyme activity were statistically significant only for
PDT dose of 6 J/cm2 in comparison to control cells
(p < 0.015).
Immunocytochemistry of iNOS. The results of
iNOS reaction are presented in the Table 1 and on
the Fig. 3. The most intensive immunocytochemical
reaction was observed 18 h after HpD-PDT (Fig. 3, c
and d). For both irradiation doses, directly after PDT
only individual cells showed the reaction (Fig. 3, b), and
in 3 h after PDT 50% of cells were stained (Table 1).
Table 1. The effect of PDT with HpD on iNOS expression in HeLa cells.
PDT dose Incubation time
after PDT, h
Staining
intensity
Immunopositive
cells,%HpD, μg/ml Irradiation, J/cm2
0 0 0 – 0
30 0 0 – 0
30 3 0 – / + single cells
6 – / + single cells
30 3 3 + 50
6 + 48
30 3 18 + + 98
6 + + 80
DISCUSSION
The cellular response to PDT is monitored by nu-
merous parameters such as the intracellular localiza-
tion of the drug, the physical and chemical properties
of the photosensitizer and the cell type. Depending on
the photosensitizer used, photodynamic treatment has
been shown to induce lesions in membranes, mito-
chondria, lysosomes or DNA [26]. Our in vitro studies
showed that the intracellular accumulation of HpD was
time dependent in HeLa cells. This is similar to our
previous study where we showed a time dependent
increase of accumulation of the fluorescent dye (Ph,
Photofrin®) in other malignant cell lines (A549, MCF-7,
Me45). Our data showed that staining was mainly
localized in intracellular compartments, but in several
cases also in mitochondrial membranes [27]. Other
study showed that about 15% of HpD was localized in
the nucleus of two cell lines: human glioma (BMG-1)
and sqamous carcinoma (4451) cell lines [28]. How-
ever, localization in the nucleus may not always lead
to cell death and can induce mutations leading to
carcinogenesis [29].
We observed that PDT- HpD induced the oxidative
stress in HeLa cells. The light dose and the time of incu-
bation after PDT affected significantly dynamic changes
in the level of oxidative stress. The photodynamically-
generated increase of MDA concentration with additional
0
5
10
15
20
25
0 h 3 h 18 h Control cells
SO
D
ac
tiv
ity
(U
/m
g
of
p
ro
te
in
)
a
b
c
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0 h 3 h 18 h Control
3 J/cm2
6 J/cm2
Control cells
0
5
10
15
20
25
30
35
40
45
50
0 h 3 h 18 h Control cells
3 J/cm2
6 J/cm2
Control cells
M
DA
c
on
ce
nt
ra
tio
n
(μ
m
ol
/l)
Th
e
le
ve
l o
f -
SH
g
ro
up
s
(μ
m
ol
/l)
*
*
*
*
*
*
*
*
*
Fig. 2. The evaluation of oxidative stress markers in HeLa cell after
photodynamic treatment: a, The production of malondialdehyde by
HeLa cells after PDT with HpD. The level of MDA was quantified at
once after photodynamic reaction and after 3 and 18 h; b, The level
of protein-associated -SH groups in HeLa cells after photodynamic
treatment with PDT. The protein degradation was quantified after
photodynamic reaction after 0, 3 and 18 h; c, The changes of SOD
activity in HeLa cells after photodynamic treatment with HpD. Cont-
rol cells were not irradiated and treated with HpD. Error bars shown
are means ± SD for n = 3 (n = 4 for C); *p < 0.05.
a
b
Fig. 1. Confocal microscopy of the distribution of HpD in HeLa
cells incubated for 4 h; a, fluorescent microscopy, b, brightfield
image (x1000)
198 Experimental Oncology 31, 195–199, 2009 (December)
protein damage (-SH groups) were observed directly
after PDT. Peroxidation of membrane lipids is known to
disrupt membrane structure and induce loss of function
which could lead to cell death. Our investigation con-
firms earlier PDT studies on different cell lines, showing
that after PDT the LPO level was increased [30, 31]. Our
results showed that LPO decreased with the increase of
incubation time after PDT and negatively correlated with
the thiol groups’ concentration.
The oxidative stress induced by PDT causes damage
to cellular macromolecules such as proteins, lipids and
nucleic acids. Among these targets, the peroxidation of
lipids is particularly damaging because the formation of
LPO products leads to a facile propagation of free radicals
and being accompanied with the oxidation of thiol groups
leads to the membrane disintegration [13].
Our results show that increasing incubation time af-
ter PDT induces the rise of iNOS expression. Inducible
NOS is a target enzyme because it is not dependent
on calcium concentration. Additionally, iNOS produces
NO much more efficiently than eNOS or nNOS, and
this efficiency allows effective treatment at low levels
of transgenic expression [32]. NO may play a role in
tumor development, tumor immune responses, me-
tastasis and apoptosis [23]. It has been demonstrated
that NO can modulate the effect of cancer therapies
that have an oxidative mechanism [33, 34].
We showed that in photodynamically treated HeLa
cells decrease of LPO induced rise of SH-groups level
and slow increase in iNOS activity. The current results
indicate the PDT-HpD influence on generation of ROS,
which are a signal for development of apoptosis or necro-
sis in human cervical carcinoma cells [4, 9]. Bar et al. [35]
investigated the response of OvBH-1 cells to Ph II-PDT.
These cells demonstrated pronounced resistance to
PDT-induced apoptosis, which was visualized by the lack
of remarkable changes in their morphological features,
DNA integrity and expression of apoptosis related pro-
teins [35]. In the present study the total intracellular SOD
activity in the cells after PDT was examined. Our results
showed that HpD-PDT induced the increase of SOD ac-
tivity. SOD is a protective enzyme and reduces oxidative
stress. Inhibition of SOD causes accumulation of cellular
O2
– and leads to free-radical-mediated damage to mito-
chondrial membranes, the release of cytochrome c from
mitochondria and apoptosis of the cancer cells [15, 36,
37]. Differences in cell susceptibility to PDT may depend
upon a protective mechanism, such as MnSOD gene
induction [38]. Other authors used ZnTM-2(3,4)-PyP4+,
that can act as a photosensitizer with efficacy compa-
rable to that of HpD in preventing cell proliferation and
causing cell death in vitro. They applied this photosensi-
tizer to colon adenocarcinoma cells which induced LPO,
membrane permeability, oxidative DNA damage, and the
activities of SOD, catalase, glutathione reductase, and
glutathione peroxidase [39]. Kasugai et al. [37] checked
the novel water-soluble Fe-porphyrins with SOD activity
in Walker 256, H-4-II-E and FR cells. It was shown that
cell death can be induced by Fe-porphyrins that affect
SOD mimic. The authors’ results suggest that for the
SOD mimic, O2
– may be applied as a target molecule to
provoke selectively cancer cell death. They intend new
metalloporphyrins possessing SOD activity as a new
class of anticancer agents. Other investigations indicate
that targeting SOD may be a promising approach for se-
a
b
c
d
Fig. 3. The expression of iNOS in (a) control HeLa cells (x1000);
(b) immediately after PDT (the radiation dose: 6 J/cm2, the HpD
concentration: 30 μg/mL) (x400); (c) 18 h post PDT (the light
dose: 3 J/cm2, the HpD concentration: 30 μg/mL) (x400); (d)
18 h post PDT (the light dose: 6 J/cm2, the HpD concentration:
30 μg/mL) (x1000)
Experimental Oncology 31, 195–199, 2009 (December) 199
lective killing of cancer cells, and that mechanism-based
combinations of SOD inhibitors (2-Metoxyestradiol)
with free-radical-producing agents may have clinical
applications [15].
Our study showed that PDT induced the increase
of SOD activity and LPO and decrease of the level of
thiol groups.
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