organisation internationale de la vigne et du vin paris 14 mars 2009 anticancer effects of grape...
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Organisation Internationale de la Vigne et du VinParis 14 mars 2009
ANTICANCER EFFECTS of GRAPE SEED EXTRACTS FROM ITALIA, PALIERI and RED GLOBE CULTIVARS, on HUMAN
COLON CANCER CELL LINES
Mariano BizzarriDept. Experimental Medicine University La Sapienza, Roma
AIM of the STUDY
1. CORRELATION between ANALYTE COMPOSITION of both SKIN and SEED EXTRACTS to SPECIFIC CULTIVARS and AGRONOMICTECHNOLOGIES
2. CELL CYCLE EFFECTS on DIFFERENT CANCER CELL LINES
3. APOPTOTIC EFFECTS ON CANCER CELL LINES
4. RECOGNIZE the APOPTOTIC PATHWAY TRIGGERED by GRAPESEED or SKIN EXTRACTS
CORRELATION between ANALYTE COMPOSITION of both SKIN
and SEED EXTRACTS to SPECIFIC CULTIVARS and AGRONOMIC
TECHNOLOGIES
CULTIVARS :
RED GLOBE
PALIERI
UVA ITALIA
LC/MS ANALYSIS of GRAPE SEED EXTRACTS
CELL CYCLE EFFECTS on DIFFERENT CANCER CELL LINES
CELL LINES
CONTROL: NORMAL HUMAN FIBROBLASTS
CANCER LINES:
BREAST CANCER- MCF7- MDA-MB231
COLON CANCER- CACO2- HCT8
HF SEMI PALIERI
0
0,1
0,2
0,3
0,4
0,5
0,6
0,7
0,8
0 h 24 h 48 h 72 h 96 h
n°
cell
x10
5 CTRL
EtOH
PAL 50 ug/mL
PAL 100 ug/mL
HF SEMI ITALIA
0
0,1
0,2
0,3
0,4
0,5
0,6
0,7
0,8
0 h 24 h 48 h 72 h 96 h
n°
cell
x 10
5 CTRL
EtOH
ITA 50 ug/mL
ITA 100 ug/mL
HF SEMI RED GLOBE
0
0,1
0,2
0,3
0,4
0,5
0,6
0,7
0,8
0 h 24 h 48 h 72 h 96 h
n°
cell
x 10
5 CTRL
EtOH
RG 50 ug/mL
RG 100 ug/mLNORMAL HUMAN FIBROBLASTS
CACO-2 SEMI ITALIA
0
5
10
15
20
25
30
35
0 H 24 H 48 H 72 H 96 H
N°
CE
LL
UL
E X
105 CNTR
ETOH
10 ug/mL
25 ug/mL
50 ug/mL
100 ug/mL
CACO2 SEMI PALIERI
0
5
10
15
20
25
30
35
0 H 24 H 48 H 72 H 96 H
N°
CE
LL
UL
E X
105 CNTR
ETOH
10 ug/mL
25 ug/mL
50 ug/mL
100 ug/mL
CaCO-2 SEMI RED GLOBE
0
5
10
15
20
25
30
35
40
45
0 h 24 h 48 h 72 h 96 h
n°
cell
x 10
5
CTRL
EtOH
10 ug/mL
25 ug/mL
50 ug/mL
100 ug/mL
HCT-8 SEMI ITALIA
0
2
4
6
8
10
12
0 h 24 h 48 h 72 h 96 h
n°
ce
llule
x 1
05 CTRL
EtOH
25 ug/mL
50 ug/mL
100 ug/mL
HCT-8 SEMI PALIERI
0
2
4
6
8
10
12
0 h 24 h 48 h 72 h 96 h
n°
ce
llule
x 1
05 CTRL
EtOH
25 ug/mL
50 ug/mL
100 ug/mL
HCT-8 SEMI RED GLOBE
0
2
4
6
8
10
12
0 h 24 h 48 h 72 h 96 h
n°
ce
llule
x 1
05 CTRL
EtOH
25 ug/mL
50 ug/mL
100 ug/mL
APOPTOTIC EFFECTS ON CANCER CELL LINES
apoptosi 24h CaCO-2 ITALIA
0
5
10
15
20
25
10ug/mL 25ug/mL 50ug/mL 100ug/mL% c
ell
An
n V
po
s/ 7
-AA
D n
eg
CTRL
CAMPTOTECINA
SEMI ITA
apoptosi 24h CaCO-2 PALIERI
0
5
10
15
20
10ug/mL 25ug/mL 50ug/mL 100ug/mL% c
ell
An
n V
po
s/ 7
-AA
D n
eg
CTRL
CAMPTOTECINA
SEMI PAL
apoptosi 24h CaCO-2 RED GLOBE
0
5
10
15
20
10ug/mL 25ug/mL 50ug/mL 100ug/mL% c
ell
An
n V
po
s/ 7
-AA
D n
eg
CTRL
CAMPTOTECINA
SEMI RG
Apoptosi 24h HCT-8 ITALIA
0
10
20
30
40
50
60
10ug/mL 25ug/mL 50ug/mL 100ug/mL
CTRL
CAMPTOTECINA
SEMI ITA
Apoptosi 24h HCT-8 PALIERI
0
10
20
30
40
50
60
70
10ug/mL 25ug/mL 50ug/mL 100ug/mL
CTRL
CAMPTOTECINA
SEMI PAL
Apoptosi 24h HCT-8 RG
0
5
10
15
20
25
30
35
10ug/mL 25ug/mL 50ug/mL 100ug/mL
CTRL
CAMPTOTECINA
SEMI RG
RECOGNIZE the APOPTOTIC PATHWAY TRIGGERED by GRAPE
SEED or SKIN EXTRACTS
AIF/tub GSE ITALIA
0
0,05
0,1
0,15
0,2
0,25
0,3
0,35
0,4
0,45
CTRL 10 ug/mL 25 ug/mL 50 ug/mL CTRL 10 ug/mL 25 ug/mL 50 ug/mL
24 h 48 h
O.D
. A
IF/
O.D
. tu
b
CACO2 CELLS
PARP/tub GSE ITALIA
0
0,2
0,4
0,6
0,8
1
1,2
1,4
CTRL 10 ug/mL 25 ug/mL 50 ug/mL CTRL 10 ug/mL 25 ug/mL 50 ug/mL
24 h 48h
O.D
. P
AR
P/
O.D
. tu
b
AIF/tub GSE PALIERI
0
0,2
0,4
0,6
0,8
1
1,2
1,4
1,6
1,8
CTRL 10 ug/mL 25 ug/mL 50 ug/mL CTRL 10 ug/mL 25 ug/mL 50 ug/mL
24 h 48 h
O.D
. A
IF/
O.D
. tu
b
PARP/tub GSE PALIERI
0
0,5
1
1,5
2
2,5
3
3,5
4
4,5
5
CTRL 10 ug/mL 25 ug/mL 50 ug/mL CTRL 10 ug/mL 25 ug/mL 50 ug/mL
24 h 48 h
O.D
. P
AR
P/
O.D
. tu
b
AIF/tub GSE RED GLOBE
0
0,1
0,2
0,3
0,4
0,5
0,6
0,7
0,8
CTRL 10 ug/mL 25 ug/mL 50 ug/mL CTRL 10 ug/mL 25 ug/mL 50 ug/mL
24 h 48 h
O.D
. A
IF/
O.D
tu
b
PARP/tub GSE RED GLOBE
0
0,2
0,4
0,6
0,8
1
1,2
1,4
1,6
1,8
2
CTRL 10 ug/mL 25 ug/mL 50 ug/mL CTRL 10 ug/mL 25 ug/mL 50 ug/mL
24 h 48 h
O.D
. A
IF/
O.D
. tu
b
AIF/tub GSE ITALIA
0
0,5
1
1,5
2
2,5
3
3,5
CTRL ITA24h
S10 ITA24h
S25 ITA24h
S50 ITA24h
CTRL ITA48h
S10 ITA48h
S25 ITA48h
S50 ITA48h
O.D
. AIF
/O.D
. tu
bHCT8 CELLS
cl-PARP/tub GSE ITALIA
0
1
2
3
4
5
6
CTRL ITA24h
S10 ITA24h
S25 ITA24h
S50 ITA24h
CTRL ITA48h
S10 ITA48h
S25 ITA48h
S50 ITA48h
O.D
. cl-
PA
RP
/O.D
. tu
b
AIF/tub GSE PALIERI
0
2
4
6
8
10
12
CTRL PAL24h
S10 PAL24h
S25 PAL24h
S50 PAL24h
CTRL PAL48h
S10 PAL48h
S25 PAL48h
S50 PAL48h
O.D
. AIF
/O.D
. tu
b
cl-PARP/tub GSE PALIERI
0
5
10
15
20
25
30
35
40
CTRL PAL24h
S10 PAL24h
S25 PAL24h
S50 PAL24h
CTRL PAL48h
S10 PAL48h
S25 PAL48h
S50 PAL48h
O.D
. cl-
PA
RP
/O.D
. tu
b
THE KINASE “CONNECTION”
PI3K/tubulin GSE PALIERI
0
0,5
1
1,5
2
2,5
3
3,5
CTRL 24 PAL S1024
PAL S2524
PAL S5024
CTRL 48 PAL S1048
PAL S2548
PAL S5048
O.D
. P
I3K
/ O
.D.
tub
uli
n
OTHER CANCERS
MDA-MB231 SEMI PALIERI
0
0,5
1
1,5
2
2,5
3
3,5
4
0 h 24 h 48 h 72 h 96 h
n°c
ellx
10
5
CTRL
EtOH
PAL 25 ug/mL
PAL 50 ug/mL
PAL 100 ug/mL
Breast cancer
PROLIFERATION CURVES
Apoptosi 24h MDA-MB231 PALIERI
0
1
2
3
4
5
6
7
8
9
10
10ug/mL 25ug/mL 50ug/mL 100ug/mL
% c
ell A
nn
V p
os/
7-A
AD
neg
CTRL
CAMPTOTECINA
SEMI PAL
APOPTOSIS
WORKING GROUP
Dep.t of EXPERIMENTAL MEDICINE
University La Sapienza, Roma
S. Dinicola
A. Cucina
A. Pasqualato
F. D’Anselmi
S. Proietti
M. Bizzarri
Dep.t of CHEMISTRYUniversity La Sapienza, Roma
A. Laganà
C. Cavaliere
Dep.t of VEGETAL BOTANICSUniversity La Sapienza, Roma
G. Pasqua
A.R. Santamaria
CRA-UTV Agricultural Research Council - Research Unit for table grapes and wine
growing in Mediterranean environment - Bari
D. Antonacci