Advances in Medical Sciences 62 (2017) 158–164

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Advances in Medical Sciences

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Original Research Article

Analysis of antiproliferative effect of Chamerion angustifolium water

extract and its fractions on several breast cancer cell lines
Audrius Maruška a,*, Rasa Ugenskienė b, Danguolė Raulinaitytė b, Elona Juozaitytė c,
Vilma Kaškonienė a, Tomas Drevinskas a, Ada Stelmakienė d, Ieva Akuneca a,
Tomas Makaravičius a, Nicola Tiso a, Violeta Bartkuvienė a, Olga Kornyšova a,
Ona Ragažinskienė d, Kristina Ramanauskienė e, Vitalis Briedis e
a
Department of Biology, Faculty of Natural Sciences, Vytautas Magnus University, Kaunas, Lithuania
b
Oncology Research Laboratory, Oncology Institute, Lithuanian University of Health Sciences, Kaunas, Lithuania
c
Oncology Institute, Lithuanian University of Health Sciences, Kaunas, Lithuania
d
Sector of Medicinal Plants, Kaunas Botanical Garden of Vytautas Magnus University, Kaunas, Lithuania
e
Faculty of Pharmacy, Lithuanian University of Health Sciences, Kaunas, Lithuania

A R T I C L E I N F O A B S T R A C T

Article history: Purpose: To evaluate the antiproliferative effect of the aerial part of Chamerion angustifolium (L.) Holub.
Received 9 January 2016 (Onagraceae) extract and its fractions in vitro. This is the first study on the anti-proliferative effect of C.
Accepted 22 August 2016 angustifolium on 3 distinct breast cancer cell lines.
Available online
Material/methods: Breast cancer cell lines MCF7, MDA-MB-468 and MDA-MB-231 were exposed to
different concentrations of the water extract of C. angustifolium, where DPPH radical scavenging activity
Keywords: was 0.018–0.443 mg/ml, expressed in rutin equivalents. Cell growth was analyzed after 24, 48 and 72 h
MCF7 cells
of incubation. Solid-phase extraction was applied for the fractionation of C. angustifolium water extract
MDA-MB-468 cells
MDA-MB-231 cells
and MDA-MB-468 cell line growth was tested using different fractions.
Fractionation Results: The concentrations corresponding to radical scavenging activity of 0.117 and 0.266 mg/ml
Oenothein B caused MCF7 cells growth inhibition, while in the samples exposed to the highest concentration
(0.355 and 0.443 mg/ml) no proliferation was register, suggesting cell death. MDA-MB-468 cell analysis
showed similar responses. MDA-MB-231 demonstrated cell growth inhibition following the exposure to
all analyzed high extract doses (0.117–0.443 mg/ml). MDA-MB-468 cells were selected to evaluate the
effect of fractions. In the samples exposed to the fraction containing the highest amount (91%) of
oenothein B, at the concentration of 0.117 mg/ml a pronounced cell growth inhibition while at higher
concentrations (0.266 and 0.443 mg/ml) no cell proliferation was observed.
Conclusions: The consumption of C. angustifolium herb can be advantageous, alongside with
conventional breast cancer treatment.
ß 2017 Published by Elsevier B.V. on behalf of Medical University of Bialystok.

1. Introduction
Chamerion angustifolium (L.) Holub. (Onagraceae), syn. Epilo-
bium angustifolium, is commonly known as willow-herb. Willow-
herb is a naturally growing plant of the Northern Hemisphere. C.
angustifolium is used in traditional medicine as a remedy for the
treatment of disorders of the prostate, kidneys, and urinary tract
* Corresponding author at: Department of Biology, Faculty of Natural Sciences,
Vytautas Magnus University, Vileikos str. 8, Kaunas LT-44404, Lithuania.
E-mail address: a.maruska@gmf.vdu.lt (A. Maruška).
[1,2], dysentery and abdominal cramps [3]. The willow-herb was
described in pharmacopoeia in 1880 [4]. Maria Treben described in
her book ‘‘Health through God’s Pharmacy (advice and experiences
with medicinal herb)’’ a few cases when the teas of Epilobium spp.
helped healing cancer [4]. A recent published study by Granica
et al. presents a comprehensive review on herbs, belonging to the
Epilobium genus, chemical composition, traditional uses, pharma-
cological activities of whole extracts of Epilobium herb or
oenothein B alone [5]. The main responsibility for the wide
spectrum of C. angustifolium biological activity is attributed to
oenothein B [5–9]. A recent study by Ramstead et al. showed that
oenothein B isolated from C. angustifolium enhances IFNg

http://dx.doi.org/10.1016/j.advms.2016.08.002
1896-1126/ß 2017 Published by Elsevier B.V. on behalf of Medical University of Bialystok.
 A. Maruška et al. / Advances in Medical Sciences 62 (2017) 158–164
production by lymphocytes and proves the immune enhancing
properties of this plant [6]. It was found that oenothein B isolated
from Epilobium spp. enhances IFNg production by NK cells and
T cells [10]. The anti-proliferative activity of willow herb extract on
prostate cancer cell lines has been investigated [7,8,11]. As
published data shows, that C. angustifolium has effect on prostate
cancer (the most common type of cancer in male), it is possible the
effect of this herb on the breast cancer cells. As breast cancer is the
most common type of cancer in female and to our knowledge, the
extract or fractions of C. angustifolium, was not tested on the breast
cancer cells. Granica et al. [5] in the review describes different
compounds identified in C. angustifolium by various scientists throw
over the world, also different and wide pharmacological activity of
the same herb. It should be considered that the accumulation of
biologically active compounds depends on the plant growing
conditions, i.e. geographic origin, climatic conditions and its ecotype
or chemotype. However sample preparation, for example extraction
conditions (solvent, temperature, time, pH, etc.), fractionation,
purification, pre-concentration and other factors, may play a very
important role on the pharmacological activity of the plants.
The aim of this study was to evaluate the antiproliferative effect
of the aerial part of C. angustifolium extract and its fractions on
three different breast cancer cell lines in vitro, namely MCF7, MDA-
MB-468 and MDA-MB-231.
To the author’s knowledge this is the first study of the anti-
proliferative effect of C. angustifolium grown on several breast
cancer cell lines.
2. Material and methods
2.1. Plant material
The raw material of C. angustifolium (willow-herb) was collected
in Medicinal plants sector ex situ (3200 m2) of the Kaunas Botanical
Garden of Vytautas Magnus University (Lithuania) in 2010 year. The
identification was carried out by Prof. O. Ragažinskienė. Voucher
specimens (VO1276) were deposited at the Herbarium of the
Kaunas Botanical Garden of Vytautas Magnus University.
The collected areal part of the plant was air-dried in a well-
ventilated shadow place and used for further analysis.
2.2. Preparation of C. angustifolium extract
Air-dry (water content ca. 9%) plant material of C. angustifolium
was ground to 1–2 mm particles. The material was mixed with water
at ratio 1:10 (w/V) and extracted using a magnetic stirrer at 4 8C
temperature. After 24 h the extract was centrifuged at 4000 rpm and
filtered. The extract was diluted 50-fold and used for the assessment
of the antiproliferative activity and the fractionation.
2.3. Fractionation of C. angustifolium extract
Fractionation of the C. angustifolium extract was carried out
using solid phase extraction. 50 ml of the extract were added into a
self-packed C18 cartridge. The cartridge was prepared by filling
10 g of adsorbent (SGX, C18, 60 mm, Separon Ltd. Praha, Czech
Republic) in a 100 ml polypropylene cartridge equipped with glass
paper filters (Whatman, UK) on both sides of the adsorbent bed.
Four fractions were washed out from the cartridge. The first
fraction was washed out using 100 ml of bidistilled water and
named fraction No. 1. Then cartridge was washed out using 100 ml
of 10% methanol and fraction No. 2 was obtained. Then cartridge
was washed out using 50 ml of 50% methanol and fraction No.
3 was obtained. The last fraction No. 4 was obtained, when
cartridge was washed with 90% methanol. In order to replace the
solvent, 30 ml of each fraction were evaporated to dryness using a
159
rotavapor Heideloph VV Micro Evaporator (Germany) equipped
with a water jet vacuum pump and dissolved in 30 ml of water.
The radical scavenging activity of the obtained fractions was
tested using DPPH (2,2-diphenyl-1-picrylhydrazyl) assay [12] and
fractions were diluted in order to obtain the same radical
scavenging activity as whole extract (approx. 7.71 mg/ml
expressed in rutin equivalents; data not shown) in order to avoid
the impact of different antiradical activity. Since fractions No.
1 and No. 4 did not show any considerable radical scavenging
activity, they were not tested for antiproliferative activity. The
diluted fractions No. 2 and No. 3 with adjusted radical scavenging
activity (ca. 7.71 mg/ml in rutin equivalents) were used for further
analysis. Furthermore, the mixture of these two fractions was
analyzed and named fraction No. 2 + 3 in the text below (equal
contribution of every fraction resulting in radical scavenging
activity of the mixture ca. 7.71 mg/ml in rutin equivalents).
2.4. HPLC analysis
The qualitative evaluation of C. angustifolium extract and its
fractions was performed using HPLC equipped with UV detector.
Separation was carried out using a reversed phase analytical
column LiChrospher 100 RP-18e, 125 mm  4.6 mm i.d. (Merck,
Germany), packed with C18 stationary phase with a particle size of
5 mm. Detection was carried out at 254 nm. The eluent consisted of
two components (A and B). Component A consisted of water with
0.05% trifluoroacetic acid additive. Component B was methanol
with 0.05% trifluoroacetic acid additive. The gradient program was
as follows: from 0 to 5 min an increase from 1% to 25%, from 5 min
to 10 min held at 25%, from 10 min to 38 min an increase to 60%,
from 38 min to 40 min an increase to 99%, from 40 min to 45 min
held at 99% of component B and then in 3 min to return to the
initial conditions. Equilibration time of 10 min was allowed before
the injections. The flow rate was 0.75 ml/min and the injection
volume was 10 mL.
2.5. Cell cultures
MCF7, MDA-MB-468 and MDA-MB-231 breast cancer cell lines
were purchased from CLS Cell Line Service (Germany). Cell growing
and freezing/thawing conditions were as those recommended by
the cell line provider and this was performed by a qualified
biologist according to the rules of good laboratory practice. Cells
were grown in DMEM:Ham’s F12 (1:1) medium (Lonza) supple-
mented with 10% FBS (Gibco), 1% glutamine (Gibco) and 100 IU/ml
penicillin-streptomycin solution (Gibco) at 37 8C, humidified and
filled with 5% CO2 air conditions. MCF7, MDA-MB-468 and MDA-
MB-231 cell lines have different molecular characteristics and
represent different breast cancer subtypes.
2.6. Procedure
For experiments 200 000 cells were plated into 35 mm
diameter Petri dishes. The next day cells were exposed to the
medium containing different concentrations of C. angustifolium
extract and its fractions. Since studied extract and its fractions
were prepared in water solution, water was also used for control
samples. The amount of water added to control samples
corresponded to the amount of aqueous extract solution used in
the experiment. The viability of breast cancer cells was assessed
using hemacytometer after 24, 48 and 72 h of incubation. The
procedure in brief: trypan blue-treated cell suspension was
applied to the hemacytometer. Live cells excluded dye and were
colorless and refractile when scored under the microscope (Fig. 1).
During the process of experimental protocol optimization,
different concentrations of extract (0.018–1.500 mg/ml, expressed
160 A. Maruška et al. / Advances in Medical Sciences 62 (2017) 158–164
Fig. 1. Live and dead MKN7 cells (live cells are colorless and refractile; dead cells are
blue).
as DPPH radical scavenging activity in rutin equivalents) and
various exposure times (24–72 h) were investigated (data not
shown). Finally, the solutions with DPPH radical scavenging
activity in between 0.018 and 0.443 mg/ml and time points of 24,
48 and 72 h were selected for the experiments as the best
representing studied effects. The experiments were not carried on
beyond 72 h time point as at that time cell culture was confluent in
the controls and samples exposed to the lowest concentration of
the studied extract.
For further analysis two fractions of the plant extract, namely
No. 2 and No. 3, and their mixture No. 2 + 3 were prepared. The
studied extract fractions were diluted to result DPPH radical
scavenging activity expressed in rutin equivalents in between
0.117 and 0.443 mg/ml.
2.7. Statistical evaluation of the data
The statistical data analyses were performed with SPSS
(Statistical Package for the Social Sciences) version 20.0 statistical
Fig. 2. The effect of C. angustifolium extract concentrations expressed in radical scavenging activity and duration of the exposure on MCF7 cell line (columns and bars represent
mean  STDEV). * – the statistically significant differences (p < 0.01) between controls and the samples exposed to various C. angustifolium extract concentrations.
software (SPSS Inc., Chicago, IL). The Dunnett (2-sided) test was
used for the comparison between the controls and the samples
exposed to the extract or its fractions. LSD (least significant
difference) test assisted the data analysis when the comparison
was made between the samples exposed to different extract
fractions. p < 0.05 was considered as a statistically significant.
3. Results
The first task of this study was to evaluate the effect of C.
angustifolium water extract on MCF7, MDA-MB-468 and MDA-MB-
231 breast cancer cell lines. The effect of different C. angustifolium
extract concentrations against tested cell lines are presented in
Figs. 2–4. The statistical data analysis demonstrating significant
differences (p < 0.01) between controls and the samples exposed
to various extract concentrations are also presented in Figs. 2–4.
The second task was to evaluate the effect of different C.
angustifolium water extract fractions (fraction with enhanced
amount of oenothein B, fraction with reduced amount of oenothein
B and mixture of both fractions) on MDA-MB-468 cell growth. This
cell line was chosen as it represents a triple negative subtype of
breast cancer, which is associated with poor prognosis. Treatment
of triple negative cancer is a considerable challenge in contempo-
rary oncology.
The results of fractionation is given in Fig. 5; while the effect of
different C. angustifolium extract fractions on MDA-MB-468 cell
line is presented in Fig. 6.
The doubling time of tested cells is summarized in Tables 1
and 2.
4. Discussion
The searching of bioactive compounds from natural sources
exhibiting a wide variety of biological activities became very
popular at the end of the last century. Natural products (herbs,
vegetables, fruits, spices, etc.) are an excellent source of complex
chemicals, having a great therapeutic potential. It was discovered
that black cohosh (Cimicifuga racemosa L.) (family Ranunculaceae)
and related Cimicifuga species have antiproliferative effect against
breast cancer [13]. Alkaloids from the seeds of Strychnos nux-
vomica L., family Loganiaceae, exhibit the anti-tumor effect on
HepG2 liver cancer cell line [14]. Patrinia scabiosaefolia Fischer,
family Valerianaceae, induces apoptosis in human breast carcino-
ma MCF-7 cells [15]. Peach and plum polyphenols inhibit the
proliferation of the estrogen-independent MDA-MB-435 breast
cancer cell line [16]. The fenugreek extract induces apoptosis of
 A. Maruška et al. / Advances in Medical Sciences 62 (2017) 158–164 161

Fig. 3. The effect of C. angustifolium extract concentrations expressed in radical scavenging activity and duration of the exposure on MDA-MB-468 cell line (columns and bars
represent mean  STDEV). * – the statistically significant differences (p < 0.01) between controls and the samples exposed to various C. angustifolium extract concentrations.

Fig. 4. The effect of C. angustifolium extract concentrations expressed in radical scavenging activity and duration of the exposure on MDA-MB-231 cell line (columns and bars
represent mean  STDEV). * – the statistically significant differences (p < 0.05) between controls and the samples exposed to various C. angustifolium extract concentrations.

Fig. 5. HPLC chromatographic profiles of C. angustifolium initial extract and obtained fractions.
162 A. Maruška et al. / Advances in Medical Sciences 62 (2017) 158–164

Fig. 6. The effect of different C. angustifolium extract fractions, concentrations expressed in radical scavenging activity and duration of the exposure on MDA-MB-468 cell line
(columns and bars represent mean  STDEV). * – the statistically significant differences (p < 0.01) between controls and the samples exposed to various C. angustifolium extract
fractions. * – the statistically significant differences (p < 0.05) between the samples exposed to corresponding concentrations of the fraction No. 3 and No. 2. & – the statistically
significant differences (p < 0.05) between the samples exposed to corresponding concentrations of the fraction No. 2 + 3 and No. 2. & – the statistically significant differences
(p < 0.01) between the samples exposed to corresponding concentrations of the fraction No. 2 + 3 and No. 3.

MCF-7 cells [17]. Nevertheless not all plants have positive effect. equivalents 0.117 and 0.266 mg/ml) of C. angustifolium the cell
For example, the study by Sebastian and Thampan demonstrated growth inhibition was observed. This might be due to the cell cycle
that the soybean extract acts as a promoter of MCF-7 cell growth delay or early induction of apoptosis. The doubling time of MCF7
[17]; Angelica sinensis (Oliv.) Diels, family Apiaceae (dong quai) and cells was 38 h in controls and 45 h in the samples exposed to the
ginseng (Panax spp., family Araliaceae), which help to release extract with the radical scavenging activity of 0.117 mg/ml
menopausal symptoms, also stimulate the growth of MCF-7 cells (Table 1). Cell doubling time was assessed with online ‘‘Doubling
[18]. Hence it is important to investigate the proliferative effect of time’’ computer program [19]. Even though the number of cells
the herbs, which are widely used in the folk medicine. was gradually increasing in the samples with 0.266 mg/ml extract
concentration (radical scavenging activity), it did not double
4.1. The effect of C. angustifolium extract on breast cancer cell lines during the experimental time (72 h). Finally, in the samples
exposed to the highest extract concentrations (radical scavenging
The data obtained from MCF7 cells suggest that the effect of the activity 0.335 and 0.443 mg/ml) the number of cells at all analyzed
analyzed extract is dose dependent (Fig. 2). In the samples exposed time points was lower (<200 000) than that at zero time point,
to the lowest concentrations of C. angustifolium (radical scavenging suggesting cell death.
activity 0.018 and 0.089 mg/ml) cell growth was similar to that of In the experiments with MDA-MB-468 cell line, samples
the control samples. However, in the samples exposed to higher exposed to the lowest concentration of extract (radical scavenging
concentrations (radical scavenging activity expressed in rutin activity 0.018 and 0.089 mg/ml) presented with the similar cell
number as controls (Fig. 3). The samples treated with the extract of
radical scavenging activity 0.117 mg/ml, demonstrated cell growth
inhibition during the whole experiment (after 24, 48 and 72 h). In
Table 1 those samples the number of cells was slowly increasing, however,
The doubling time of breast cancer cells exposed to C. angustifolium extract.
during the experimental time (72 h) it did not reach the doubling
Cell line Doubling time (h) point. As far as the samples exposed to higher extract concentra-
Control 0.117 mg/mla 0.266 mg/mla
tions (radical scavenging activity 0.266, 0.354 and 0.443 mg/ml)
are concerned, they presented with the number of cells which was
MCF-7 38 45 –
always lower (<200 000) than it was at zero time point, indicating
MDA-MB-468 45 – –
MDA-MB-231 38 41 59 cell death. The observed effect was more pronounced at the highest
a
extract dose (radical scavenging activity 0.443 mg/ml) and at 72 h
Samples exposed to the extract with radical scavenging activity expressed in
rutin equivalents.
time point.
The C. angustifolium extract also showed a dose dependent
effect on MDA-MB-231 breast cancer cell line (Fig. 4). Similar to the
Table 2 above mentioned observations, the lowest extract doses (radical
The doubling time of MDA-MB-468 cell line exposed to different C. angustifolium scavenging activity expressed in rutin equivalents 0.018 and
extract fractions.
0.089 mg/ml) had no impact on MDA-MB-231 cell growth.
Fraction Doubling time (h) Whereas in the rest of the samples a pronounced cell growth
Control 0.117 mg/mla 0.266 mg/mla inhibition was observed and it was extract dose and exposure time
dependent (more obvious at higher doses and at 72 h time point).
No. 2 45 – NA
No. 3 45 61 –
MDA-MB-231 cell doubling time in control samples was 38 h;
No. 2 + 3 45 – NA while in the samples, exposed to the extract at the concentration of
a 0.266 mg/ml, the doubling time was 59 h (Table 1). The samples
Samples exposed to the extract with radical scavenging activity expressed in
rutin equivalents. with the highest extract concentration (radical scavenging activity
NA – not applicable as no cell division was observed. 0.354 and 0.443 mg/ml) did not get to the doubling point during
 A. Maruška et al. / Advances in Medical Sciences 62 (2017) 158–164
72 h. The results form statistical data analysis demonstrating
significant differences (p < 0.01) between controls and the
samples exposed to various extract concentrations are presented
in Figs. 2–4.
4.2. The effect of different C. angustifolium extract fractions on cell
growth
Primarily, an HPLC analysis of the C. angustifolium fractions was
performed in order to reveal whether the prepared fractions differ
by their qualitative composition. The HPLC chromatograms of the
fractions are presented in Fig. 5. It is evident, that oenothein B is
the most abundant compound among the phenolic compounds in
the fraction No. 2 (91% of all HPLC-UV chromatogram peak area),
while the area of peak of oenothein B was almost 6 times smaller in
the fraction No. 3, where other peaks of the phenolic compounds
constituted the majority of the total peak area (89%). The
quantitative analysis of separated compounds and identification
of other phenolic compounds were out of the scope of this study.
Literature data shows that oenothein B concentration in C.
angustifolium is 225.8 mg/g, while other flavonoids only 13.4 mg/g
[20]. The fraction of other flavonoids of Epilobium species consists
of myricetin, quercetin, kaempferol and their various glycosides
[21,22]. Ferulic, gallic, protocatechuic, cinnamic, caffeic, gentisic
and chlorogenic acids were also identified in the extract of C.
angustifolium by Hiermann and Radl using capillary zone
electrophoresis [23]. Granica et al. [5] in their review described
five groups of chemical compounds detected in Epilobium spp.:
flavonoids (myricetin, quercetin, kaempferol and their glycosides),
phenolic acids and their derivatives (ellagic acid and its
derivatives, chlorogenic acids, benzoic acid derivatives with gallic
acid, trans-cinnamic acid derivatives), tannins and related com-
pounds (oenothein A, oenothein B, and gallotannins), steroids and
triterpenes, and other compounds.
The aim of fractionation was to obtain qualitatively different
fractions containing predominantly oenothein B and other
phenolic compounds in separate fractions. Additional fraction
was a blend of two qualitatively different fractions of equal radical
scavenging activity. For comparative reasons all the fractions were
diluted to yield the same radical scavenging activity (7.71 mg/ml).
Fig. 5 presents that the fractionation of C. angustifolium extract was
successful.
The effect of different C. angustifolium extract fractions on MDA-
MB-468 cell line is presented in Fig. 6. The presented results in each
plot come from 4 to 6 independent measurements.
Two different extract fractions namely No. 2 and No. 3 and their
mixture No. 2 + 3 were prepared adjusting their radical scavenging
activity to that of initial extract (7.71 mg/ml). Fraction No.
2 showed the most prominent effect in comparison to the others
on the selected cell line. In the samples exposed to the lowest
extract concentration (radical scavenging activity 0.117 mg/ml),
cell growth inhibition was observed as the number of cells has only
slightly increased during the whole experiment. Furthermore, in
the samples with higher extract concentrations (radical scaveng-
ing activity 0.266 and 0.443 mg/ml) the number of cells was
always lower (<200 000) than it was at zero time point, indicating
cell death. That was more protrusive in the samples with the
highest extract concentration (radical scavenging activity
0.443 mg/ml) and at 72 h time point.
The fraction No. 3 showed weaker effect on MDA-MB-468 cells
than the fraction No. 2. In summary, at the radical scavenging
activity of 0.117 and 0.266 mg/ml, cell growth inhibition was
observed. MDA-MB-468 cell doubling time was 45 h in controls
and 61 h in the samples exposed to the fraction No. 3 at the
concentration of 0.117 mg/ml (Table 2). In the samples exposed to
the highest concentration of this fraction (radical scavenging
163
activity 0.443 mg/ml) the number of cells was always lower
(<200 000) than that at zero time point, indicating cell death.
The experiments with the fraction No. 2 + 3 showed average
results as its effect on MDA-MB-468 cell line was weaker than
that of the fraction No. 2, but stronger than the effect of the
fraction No. 3 (Fig. 5). In the samples with the lowest fraction
No. 2 + 3 concentration (radical scavenging activity 0.117 mg/ml),
cell growth inhibition was registered as the number of cells only
slightly increased during 72 h. In the samples exposed to the
higher concentrations (radical scavenging activity 0.266 and
0.443 mg/ml) of the fraction No. 2 + 3, the number of cells was
always lower (<200 000) than that at zero time point, indicating
cell death. The statistically significant differences between controls
and the samples exposed to the different extract fractions and in-
between fractions are presented in Fig. 6.
5. Conclusions
The results indicate that C. angustifolium extracts at the radical
scavenging activity expressed in rutin equivalents range between
0.266 and 0.443 mg/ml alters MDA-MB-231 and MCF7, MDA-MB-
468 cell growth. The extract concentration resulting radical
scavenging activity 0.443 mg/ml had the highest effect on MCF7
and MDA-MB-468 cells after 72 h. The MDA-MB-468 cell line was
selected for the evaluation of C. angustifolium fractions. Both cell
growth inhibition and cell number reduction, suggesting cell
death, were observed in the samples exposed to the fraction No.
2 containing the highest amount (91%) of oenothein B.
Acknowledgement
Authors acknowledge donation of C18 Separon adsorbent by
Prof. Dušan Berek.
Conflict of interest
The authors declare that they have no conflict of interest.
Financial disclosure
The study was financially supported by the Research Council of
Lithuania (grant no. MIP 084/2012 ONKOFITAS).
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