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Enhanced Chemopreventive Effect
by Combining Quercetin and Green
tea in Prostate Cancer
Piwen Wang, MD, PhD
Assistant Professor, Division of Cancer Research and Training
Charles R. Drew University of Medicine and Science
(Cancer Facts & Figures 2015)
(Cancer Facts & Figures 2015)
Age-Standardized Prostate
Cancer Incidence and Mortality
Rates by World Area. Source:
GLOBOCAN 2008.
(Jemal A, et al. CA CANCER J CLIN 2011;61:69–90)
Prostate cancer
Prostate cancer: Science and clinical practice. Edited by: J Mydlo & C Godec. 2013)
Tea
From the plant Camellia sinensis
Green tea (GT):
- Not fermented
- Polyphenol monomer (EC, EGC,ECG, EGCG)
Oolong tea:
- Partially fermented
Black tea:
- Fully fermented
- Dimers and polymers
(theaflavins, thearubigins)
(Alipoor, et al. Asian J Clin Nutr. 2012, 4 (1): 1-15
GT and GT polyphenols
Anti-carcinogenesis
- Skin, lung, oral cavity, esophagus, stomach,
intestine, colon, liver, pancreas, bladder,
mammary gland, prostate
Diabetes and obesity
Cardiovascular diseases
Neurodegenerative diseases
Osteoporosis
(Yang CS, et al. Nature Reviews Cancer 2009, 9, 429-439)
Possible targets for the cancer preventive activity of (-)-epigallocatechin-3-gallate (EGCG).
Pooled odds ratio (OR) of GT consumption (highest
versus non/lowest) and risk of prostate cancer
(Zheng J. et al. 2011 Nutr and Cancer 63(5):663-672)
Intervention studies: GT and prostate cancer
Tea, Country Effect Reference
GT extract,
Italy,
600 mg
Reduced progression from
HG-PIN to prostate cancer:
Control 30%
GTE 3%
Bettuzzi S et al.
2006
GT extract, USA,
800 mg EGCG
(single arm)
Decreased serum PSA, HGF,
VEGF in prostate cancer
patients
McLarty J et al.
2009
GT extract, USA,
800 mg EGCG
(placebo
controlled)
A decreasing trend of PSA,
in prostate cancer patients
Nguyen MM et al.
2012
Screening
Randomize of eligible subjects
3-8 weeks intervention
Placebo (N=33) GT (N=34)
Urine
polyphenol
metabolites
8-OHdG
Serum
polyphenol
metabolites
ex vivo
bioassay
PSA
Prostate
polyphenol
metabolites
Prostate
proliferation
apoptosis
ROS
Bax/Bcl-2
NFkB
BT (N=26)
A phase II clinical trial of tea in prostate cancer
(Henning. et al. 2015 Prostate 75:550-559)
(Henning. et al. 2015 Prostate 75:550-559)
(Henning. et al. 2015 Prostate 75:550-559)
Limitations of GT in chemoprevention
EGCG 4"-MeEGCG ECG
Co
nc
en
tra
tio
n (
pm
ol/g
tis
su
e)
0
5
10
15
20
25
30
35
0h 24h 48h 72h
Ce
ll v
iab
ilit
y (
% o
f c
on
tro
l)
0
20
40
60
80
100
120
NT
4"-MeEGCG 40uM
EGCG 40uM
4"-MeEGCG 80uM
EGCG 80uM
a
ab
ab
abcd
ab
ab
abcd
abc
Low bioavailability of GT polyphenols
Extensively methylation in vivo
(Wang P, et al. 2010 Cancer Prev Res 3(8):985-93)
Catechol-O-methyltransferase (COMT)
CH3
EGCG
Quercetin (Q)
A flavonoid commonly found in fruits and vegetables - red
onions, apples, broccoli, berries, and tea
A natural inhibitor of catechol-O-methyltransferase (COMT) and
multidrug resistance-related proteins (MRPs)
Quercetin in combination with EGCG in vitro
– cellular absorption
EGCG 80uMQuer 5uM
Quer 10uM
EGCG + Quer 5uM
EGCG + Quer 10uM
Co
ncen
trati
on
(p
mo
l/m
g c
yto
so
lic p
rote
in)
0
100
200
300
400
500
600
700
800
900 EGCG
4"-MeEGCG
Quercetin
Isorhamnetin
a
a b
b
b
a
c
c
c
c
d
d
dd
LNCaP
EGCG 80uMQuer 10uM
Quer 20uM
EGCG + Quer 10uM
EGCG + Quer 20uM
Co
nc
en
tra
tio
n (
pm
ol/m
g c
yto
so
lic
pro
tein
)
0
100
200
300
400
500
600
700
800
EGCG
4"-MeEGCG
Quercetin
Isorhamnetin
a
a
a
b b
b
c
c
c
c
d
d
d
d
PC-3
The groups with the same superscript letter represent significant difference between groups (P<0.05)
(Wang P, et al. 2012 Nutr Cancer 64(4): 580-587)
Quercetin in combination with EGCG in vitro
– Antiproliferative effect
0 24 48
Ce
ll v
iab
ilit
y (
% o
f c
on
tro
l)
0
10
20
30
40
50
60
70
80
90
100
110
NT
EGCG 40uM
Quer 5uM
Quer 10uM
EGCG 40um + Quer 5uM
EGCG 40um + Quer 10uM
a
abcd
a
a
abc
abcd
aabcd
LNCaP
Time (Hours)
(Wang P, et al. 2012 Nutr Cancer 64(4): 580-587)
0 24 48
Cell v
iab
ilit
y (
% o
f co
ntr
ol)
0
10
20
30
40
50
60
70
80
90
100
110
NT
EGCG 40uM
Quer 10uM
Quer 20uM
EGCG 40um + Quer 10uM
EGCG 40um + Quer 20uM
abc
abc
ab
ab
abcd
abcd
PC-3
Time (Hours)
Compared to – a, control (NT); b, EGCG; c, Q 5umol/L; d, Q 10umol/L, p<0.05
Quercetin in combination with EGCG in vitro
– COMT inhibition
Time (Hours)
2 24 48
Van
illic a
cid
fo
rmed
(n
mo
l/h
/mg
pro
tein
)
0
10
20
30
40
50
60
70
80
90
NT
EGCG 40uM
Quer 10uM
Quer 20uM
EGCG 40uM + Quer 10uM
EGCG 40uM + Quer 20uM
aa
b
c
d
e
a a
b
c
d
e
ac
b
d
e
f
NT EGCG
40 μM
Quer
10 μM
EGCG 40 μM+
Quer 10 μM
48h
COMT
β-actin 24h
COMT
β-actin
Time (Hours)
24 48
CO
MT
co
ncen
trati
on
(re
lati
ve t
o N
T)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
NT
EGCG 40uM
Quer 10uM
EGCG 40uM + Quer 10uM
a a
abc
a
abc
(Wang P, et al. 2012 Nutr Cancer 64(4): 580-587)
0h 24h 48h
Cell v
iab
ilit
y (
% o
f co
ntr
ol)
0
10
20
30
40
50
60
70
80
90
100
110
NT
EGCG 40uM
Quer 10uM
Quer 20uM
EGCG 40um + Quer 10uM
EGCG 40um + Quer 20uM
a
a
abcd
a
abc
abcd
abcd
abcd
A549 (lung cancer)
0h 24h 48h
Cell v
iab
ilit
y (
% o
f co
ntr
ol)
0
10
20
30
40
50
60
70
80
90
100
110
NT
EGCG 40uM
Quer 10uM
Quer 20uM
EGCG 40um + Quer 10uM
EGCG 40um + Quer 20uM
a
abc
abcd
aaa
abcd
abcd
786-O (kidney cancer)0h 24h 48h
Ce
ll v
iab
ilit
y (
% o
f c
on
tro
l)
0
10
20
30
40
50
60
70
80
90
100
110
NT
EGCG 40uM
Quer 10uM
Quer 20uM
EGCG 40um + Quer 10uM
EGCG 40um + Quer 20uM
abcd
abcda
aa
a
abc
abcd
Hep G2 (liver cancer)
Increased anti-proliferative effect by EGCG + Q in lung, liver
and kidney cancer cells
(Wang P, et al. 2012 Food Funct.3(6):635-42)
Xenograft SCID mouse model
0.5 million LAPC4 cells
When tumor was palpable - start of intervention
Weeks 0 1 2 3 4 5 6 7
Quercetin in combination with GT in SCID mice
GT (Celestial Seasonings, Boulder,
CO): 1 tea bag brewed in 240 ml
boiling water for 5min; 0.07% GT
polyphenols
Q (Sigma-Aldrich): 0.4% in diet EGC EGCG EC ECG
Co
ncen
trati
on
(m
g/L
wate
r)
0
100
200
300
400
500
AIN 93G+Quercetin
Xenograft tumor growth
Compared to – a, control group; b, Q group; c, GT group, p<0.05
Time (Weeks)
0 1 2 3 4 5 6 7
Tu
mo
r vo
lum
e (
cm
3)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
Control
GT
Q
GT + Q
abc
a
a
a
a
abc
a
Tum
or
inocula
tion
Tre
atm
ent
sta
rted (21%)
(16%)
(45%)
(Wang P, et al. 2014 J Nutr Biochem 25(1):73-80)
Food and water consumption and body weight
Food WaterDa
ily c
on
su
mp
tio
n o
f fo
od
(g)
an
d w
ate
r (m
l) p
er
mo
use
0.0
2.0
4.0
6.0
8.0
Control
GT
Q
GT+Q
Weeks
0 1 2 3 4 5 6 7
mo
us
e w
eig
ht
(g)
16
18
20
22
24
26
Control
GT
Q
GT+Q
GT Q GT+Q
Concentr
ation (
pm
ol/g tis
sue)
0
10
20
30
40
50
60
70
80200
300
400
500
600
700EGCG
4"-MeEGCG
EGC
ECG
EC
quercetin
isorhamnetin
*
*
*
*
*
(Wang P, et al. 2014 J Nutr Biochem 25(1):73-80)
GT+Q increased the bioavailability and decreased
methylation of GT polyphenols in tumor tissues
GT Q GT+Q
Co
nc
en
tra
tio
n (
pm
ol/g
tis
su
e)
0
50
100
150
200
300
350
400
EGCG
4"-MeEGCG
EGC
ECG
EC
quercetin
isorhamnetin
Lung
*
**
*
*
*
GT Q GT+Q
Conce
ntr
atio
n (
pm
ol/g
tis
sue)
0
50
100
150
200
250
300
400
450 EGCG
4"-MeEGCG
EGC
ECG
EC
quercetin
isorhamnetin
Kidney
*
**
*
*
*
GT Q GT+Q
Co
nce
ntr
atio
n (
pm
ol/g
tis
sue)
0
100
200
300
400
500900
950
1000
1050
1100
EGCG
4"-MeEGCG
EGC
ECG
EC
quercetin
isorhamnetin
Liver
* *
*
* compared to individual treatment, p<0.05
(Wang P, et al. 2012 Food & Function 3: 635-642 )
GT+Q increased the bioavailability and decreased
methylation of GTPs in other tissues
COMT DNMT AR PSA VEGF
Pro
tein
conce
ntr
atio
n (
rela
tive t
o c
ontr
ol)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
Control
GT
Q
GT+Q
* *
** *
**
***
*
*
**
Modulation of protein marker expression
GT Control GT+Q Quercetin
COMT
GAPDH
DNMT
AR
PSA VEGF
(Wang P, et al. 2014 J Nutr Biochem 25(1):73-80)
Inhibition of proliferation marker-Ki67
(Wang P, et al. 2014 J Nutr Biochem 25(1):73-80)
GT+Q in combination with drugs for
chemotherapy of advanced prostate cancer
GT+Q+Docetaxel: Enhanced inhibition of PC-3 tumor growth
Data are presented as mean±SE. Different letters at each time point indicate statistically
significant difference between groups, p<0.05 (manuscript submitted)
Weeks
0 1 2 3 4 5 6 7 8 9 10 11
Tum
or
volu
me (
cm
3)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
Control
GT+0.4%Q
Doc 5
GT+0.4%Q+Doc 5
Doc 10
Tu
mo
r in
ocu
latio
n
Tre
atm
en
t sta
rte
da
ab
bcc
c
a
ab
b
b
b
a
a
ab
b
b
a
b
b
a
b
b
La
st
Do
c in
jectio
n
(100%)
(38%)
(34%)
Summary and Conclusion
Quercetin in combination with GT significantly enhances the
anti-carcinogenic effect in vitro and in vivo
The combined effect was associated with an increased
bioavailability and decreased methylation of GT polyphenols
These results warrant future studies to confirm the synergistic
effect of the combination of GT and quercetin in humans
In addition, this combination may improve clinical application
in treatment of advanced prostate cancer by sensitizing tumor
cells to chemotherapy
Acknowledgement
Dr. Jay Vadgama (Charles R. Drew University)
Drs. Susanne Henning, David Heber (UCLA Center for
Human Nutrition), Jonathan W. Said, Clara E. Magyar, Ngan
Doan (UCLA Department of Pathology)
Grant supports from the National Institutes of Health [NIH, NCI, NIMHD,
NCATS; Grant Nos. U54 CA143931-01, U54MD007598,
UL1TR000124 (J.V. Vadgama)], and NIH/National Center for Advancing
Translational Sciences UCLA CTSI [Grant No. KL2TR000122 (P.
Wang)]