Article ID: WMC001504

2046-1690

Protective Role Of Quercetin: Antioxidants May

Protect Cancer Cells From Apoptosis And Enhance
Cell Durability
Corresponding Author:
Dr. Zafer Akan,
Assistant Professor Dr. , Yuzuncu Yl University Deparment of Biophysics, Yuzuncu Yl University Deparment of
Biophysics, 65100 - Turkey
Submitting Author:
Dr. Zafer Akan,
Assistant Professor Dr. , Yuzuncu Yl University Deparment of Biophysics, Yuzuncu Yl University Deparment of
Biophysics, 65100 - Turkey

Article ID: WMC001504

Article Type: Research articles
Submitted on:24-Jan-2011, 01:31:05 PM GMT

Published on: 24-Jan-2011, 08:22:44 PM GMT

Article URL: http://www.webmedcentral.com/article_view/1504

Subject Categories:APOPTOSIS
Keywords:Quercetin, cancer, apoptosis, prevention, cell viability
How to cite the article:Akan Z , Garip A i. Protective Role Of Quercetin: Antioxidants May Protect Cancer Cells
From Apoptosis And Enhance Cell Durability . WebmedCentral APOPTOSIS 2011;2(1):WMC001504
Source(s) of Funding:
This project was supported by Marmara University scientific research project unit (BAPKO) and The Scientific
and Technological Research Council of Turkey (TUBITAK)
Additional Files:
Title page Quercetin Zafer Akan

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Protective Role Of Quercetin: Antioxidants May

Protect Cancer Cells From Apoptosis And Enhance
Cell Durability
Author(s): Akan Z , Garip A i

Abstract
Quercetin is one of the most abundant dietary
flavonoids widely present in many fruits and
vegetables. Previous in vitro studies has shown that
quercetin acts as an antioxidant and anti-inflammatory
agent and it has potent anticarcinogenic properties as
apoptosis inducer.
In this study we examined apoptotic effects of
quercetin on the K562 erythroleukemia cell line. K562
cells were induced to apoptosis by hydrogen peroxide
and quercetin. Cell viability and apoptosis level were
assessed by annexin V and PI staining method using
flow cytometry. Viability of K562 cells was increased
by low dose of quercetin (5-100 M) during 3 hours.
High dose of quercetin at toxic doses (100-500 M)
during 24 hours resulted with decrease of K562 cell
viability as expected (P2O2 (150, 300, 600 M). K562
cells were protected from H2O2induced apoptosis by
the Quercetin (P
As indicated in previously studies, reduction of
superoxides by the free radical scavengers such as
quercetin could be beneficial for prevention of cancer
but consumption of flavonoid free radical scavengers
during cancer treatment may weaken effect of the
chemotherapeutics and radiotherapy.

Introduction
Reactive oxygen species (ROS) are highly reactive
molecules generated predominantly during cellular
respiration and normal metabolism. These side
products can damagemany biological molecules; they
can change protein functions, damage DNA material
and cause lipid peroxidation of cell membranes (1). In
general, the reducing environment inside cells helps to
prevent free radical mediated damage. This reducing
environment is maintained by the action of antioxidant
enzymes and substances, such as superoxide
dismutase (SOD), catalase, glutathione peroxidase,
glutathione, vitamins C and E (2, 3).
Oxidative stress as a dysbalance between the
production of free radicals and antioxidant system of
the cells is involved in the pathogenesis of the deadly

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diseases including cancer and cardiovascular

diseases (4). Excessive production of ROS may lead
to programmed cell death and may accelerate the
process of aging (5).
For a long time, reactive oxygen species (ROS) have
been considered harmful mediators of inflammation
owing to their highly reactive nature. However, there
are an increasing number of findings suggesting that
ROS can play role in anti-inflammatory and prevent
autoimmune responses (6).
Neutralization of free radical is important to prevent
the damage of cells, tissues and organs. Control of
free radical neutralizations are under the control of
internal antioxidant enzymes such as SOD, NADH,
NADPH (4). Destruction of balance between the
oxidant and antioxidants depends on age,
environmental conditions and some mutations. Uses
of herbal sourced ingredients to improve balance of
antioxidant system are common way, between the
public.
Flavonoids are polyphenolic compounds that occur
ubiquitously in foods of plant origin (7). It has been
reported that some flavonoids such as rutin
(quercetin-3-rutinoside) and quercetin shows
antioxidant activity (8).
Quercetin is one of the most abundant dietary
flavonoids. It can be found in apples, black, green and
buckwheat tea, onions, raspberries, red grapes,
cherries, citrus fruits as well as in some well known
medical plants (ginkgo biloba, cranberries and St
Johns wort) (9, 10).
In previous in vitro studies has shown that quercetin
acts as an antioxidant and anti-inflammatory agent
and that it has potent anticarcinogenic properties as
apoptosis inductor.
In the normal conditions, herbal sourced antioxidants
intake could be beneficial for cancer prevention (11,
12), coronary artery disease prevention (13) and
moderation of aging process. But what will happen if
cancer patient takes artificial herbal sourced
antioxidants such as quercetin which is abundantly
present in many herbs, fruit and grain includes. Will it
facilitate anticancer therapy or not?
To examine this, K562 erythroleukemia cell line was
induced to apoptosis by hydrogen peroxide and effect
of quercetin over the apoptosis and viability of K562

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cells was observed.

Methods
Cell Culture
K562 cells (Erythroleukemia cell line derived from a
chronic myeloid leukemia patient in blast crisis) were
incubated at 37C in RPMI 1640 medium
(Sigma-Aldrich Chemie GmbH, Taufkirchen, Germany)
containing 10% FCS (Gibco-Invitrogen Ltd., Paisley,
Scotland) in a humidified 5% CO2 atmosphere. Cell
count was made with trypan blue. Viability of K562
cells were 85-90%.
Apoptosis induction by quercetin
Effect of quercetin (Sigma-Aldrich, St. Louis, MO, USA)
on the cell viability of K562 was evaluated with
different Quercetin concentration (5-500 M) through 3
hours with Flow cytometer (Becton-Dickinson, San
Jose, CA, USA). Toxic dose of Quercetin
concentrations (100-500 M) was evaluated through
24 hours.
Apoptosis induction by hydrogen peroxide
K562 cells were induced to apoptosis by with
hydrogen peroxide (H2O2 Riedel de Haen, D3016,
Seelzel, Germany) in different concentration (150, 300,
600 M) and protective effect of Quercetin (100 M)
was analyzed through 24 hours. For this aim,
protective dose of Quercetin (100 M) was added on
to K562 cell (106 cells in 10 mL RPMI 1640, %10 FCS,
penicilin/streptavidin), then hydrogen peroxide were
added.
Annexin V staining assay
After induction with H2O2 and Quercetin, K562 cells
were washed twice with PBS and suspended in 1x
?binding buffer (10 mM HEPES, 140 mM NaCl, and 5
mM CaCl2 at pH 7.4) at a concentration of approx. 1 x
105 cells/ml. 5 ml of Annexin V-FITC and 10 ml of
propidium iodide was added to both control and
induced cell suspension. After incubation at room
temperature for 10 minutes at dark, the fluorescence
of the cells was determined immediately with a flow
cytometer (FACS Calibur, Becton-Dickinson, San Jose,
CA, USA).
Statistical Analysis:
Measurements were repeated more than 4 times and
student t-test was used for statistical analysis.

Results
For the time-course and dose-response experiments,
K562 erythroleukemia cells were treated with 5, 10, 20,

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40, 80, 100, 200, 300, 400 and 500 M of quercetin for
3. Cell viability was assessed by Annexin V/PI flow
cytometry assay. Figure 1 shows that quercetin (In the
5-200 M doses for 3 hours) caused slight increase of
K562 cells viability (Figure 1) but high doses of
quercetin (over than 200 M) caused to increase of
apoptotic cell death (P
For second experiment K562 cells were treated with
100, 200, 300, 400 and 500 M of quercetin for 24 h.
Toxic doses of quercetin (doses higher than 100 M,
24 hours) resulted with decrease of K562 cell viability
as expected (P
In third probe, hydrogen peroxide (H2O2) was applied
in dose 150, 300 and 600 M for 24h. Kind of ROS,
hydrogen peroxide (H2O2) induced apoptosis of K562
cells had been shown previously and similar results
was obtained in our study (P
To determine preventive role of quercetin (free radical
scavenger flavonoid) against to H2O2 (ROS) induced
apoptosis, 100 M of quercetin was added into cell
culture medium (106 cell in the 10 ml RPMI 1640
include %10 FCS, penicillin-streptomicin), 20 minute
before the 150, 300, 600 M H2O2treatment. K562
cells viability were maintained by the Quercetin from
the H2O2 induced apoptosis (Figure 3), (P

Discussion
In previous studies quercetin was implied as an
apoptotic activator and antioxidant as well as
protective agent against various types of cancer (14).
It was shown that quercetin can inhibit proliferation of
tumor cells and reduce the number of aberrant crypt
foci in colon tumors (15) that it can facilitate
programmed cell death in lung carcinoma (16) and
colonorectal tumor cells (17). Other researchers had
showed that quercetin acts as a free radical scavenger
and protects cells from oxidizier molecules (18, 19). As
known, oxidative stress and free radicals are important
activators for apoptosis (20).
On the other hand, studies conducted in last ten years
showed that quercetin can act as an antiapoptotic
agent as well (21). In a few studies it has been shown
that quercetin can partially prevent H2O2 inducedapoptosis (22) and it was suggested that
protective effects of quercetin against oxidative injuries
of some cells may be achieved via modulation of
mitochondrial dysfunction and inhibition of caspase
activity (23).
In this study, our results showed that higher dose of
quercetin than 200 M reduce K562 cell viability
(which considered as toxic doses by Cao et al, 2007
(24)) but low doses of quercetin (2O2.

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Free radical generation and inactivations are under

the strict control of oxidoreductive reactions.
Excessive superoxide production can accelerate cell
death, DNA damage and can lead to cancer, also
excessive inactivation of superoxides can disturb
apoptosis signals. Inactivation of radiation and
environmental sourced superoxides by the herb
sourced flavonoid free radical scavengers seems
beneficial for cancer prevention (11) but if cancer is
already present, as known as free radicals are
important for apoptosis signals. For this reason,
uncontrolled consumption of flavonoid free radical
scavengers such as quercetin during chemothrapy
may weaken the effect of chemotherapeutics and
radiotherapy rather then help it.
It has been theorized that cancer risk reduction may
be achieved by greater consumption of
phytochemical-rich fruits and vegetables (25, 20).
However, results of our research suggest that
antioxidants with free radical scavenger properties can
interfere and attenuate success of anticancer therapy
so antioxidant intakes should be strictly controlled in
the cancer diagnosed patients.

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Illustrations
Illustration 1
Figures and legends

Figure captions:

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Figure 1: Effect of quercetin over the viability of K562 cells for 3 hours. Nonapoptotic cell rate
(% Cell viability) was assessed by Annexin V/PI flow cytometry assay. * P<0.05, ** P<0.01,
***P<0.001

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Figure 2: Effect of quercetin toxic doses over the K562 cells viability for 24 hours. Nonapoptotic
cells rate (% Cell viability) was assessed by Annexin V/PI flow cytometry assay. * P<0.05, **
P<0.01, ***P<0.001

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