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Curcumin : A Potent Inhibitor of Galectin-3 Expression

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The expression of galectin-3, a b-galactoside binding lectin, was found to be affected by different kinds of stressors, and is strongly modified in numerous physiological and pathophysiological conditions. Although no precise regulatory mechanisms of galectin-3 expression are unraveled, transcription factors AP-1 (activator protein 1) and NF-kB (nu- clear factor kappa B) play an important role in these processes. Activities of both tran- scription factors are affected by curcumin, a biologically active compound extracted from rhizomes of Curcuma species. We have analyzed the impact of curcumin on the expression of galectin-3 in glioblastoma cells under basal conditions and under stress invoked by the cell exposure to alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and ul- traviolet C (UV-C) light. Galectin-3 level was measured by western-blot technique using M3/38 monoclonal antibody. Curcumin has decreased the basal level of galectin-3, while the pretreatment of cells with curcumin has considerably reduced the inducible effect of UV-C radiation and abolished the inducible effect of alkylating agent. Thus, curcumin has been identified as a potent inhibitor of galectin-3 expression.
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JERKA DUMI] et al.: Curcumin – Inhibitor of Galectin-3 Expression, Food Technol. Biotechnol. 40 (4) 281–287 (2002)
281
UDC 574.979.4:578.242.44
original scientific paper
ISSN 1330-9862
(FTB-1183)
Curcumin – A Potent Inhibitor of Galectin-3 Expression
Jerka Dumi}*, Sanja Dabeli} and Mirna Flögel
Department of Biochemistry and Molecular Biology, Faculty of Pharmacy and Biochemistry,
University of Zagreb, Ante Kova~i}a 1, HR-10000 Zagreb, Croatia
Received: October 3, 2002
Accepted: November 7, 2002
Summary
The expression of galectin-3, a b-galactoside binding lectin, was found to be affected
by different kinds of stressors, and is strongly modified in numerous physiological and
pathophysiological conditions. Although no precise regulatory mechanisms of galectin-3
expression are unraveled, transcription factors AP-1 (activator protein 1) and NF-kB (nu-
clear factor kappa B) play an important role in these processes. Activities of both tran-
scription factors are affected by curcumin, a biologically active compound extracted from
rhizomes of Curcuma species. We have analyzed the impact of curcumin on the expression
of galectin-3 in glioblastoma cells under basal conditions and under stress invoked by the
cell exposure to alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and ul-
traviolet C (UV-C) light. Galectin-3 level was measured by western-blot technique using
M3/38 monoclonal antibody. Curcumin has decreased the basal level of galectin-3, while
the pretreatment of cells with curcumin has considerably reduced the inducible effect of
UV-C radiation and abolished the inducible effect of alkylating agent. Thus, curcumin has
been identified as a potent inhibitor of galectin-3 expression.
Key words: galectin-3, curcumin, UV light, alkylating agent MNNG
Introduction
Galectin-3, a 32 kD lectin that specifically recognizes
Curcumin [1,7-bis(4-hydroxy-3-methoxyphenyl)-
b-galactoside structures of glycans, plays a pivotal role
-1,6-heptadien-3,5-di-one] is a natural, biologically active
in several biological processes (1) and its ubiquitous lo-
compound extracted from rhizomes of Curcuma species
calization (in the nucleus and cytoplasm, on the cell
that gives the golden-yellow color and unique flavor to
membrane and in the extracellular matrix) only con-
curry. In Indian medicine it has been known for centu-
firms its diverse functions, including modulation of cell
ries, while Western medicine discovered only lately its
adhesive properties (2–4) and regulation of cell motility
anti-inflammatory, anti-oxidative, and cytostatic proper-
(5). Galectin-3 shows anti-apoptotic and growth-enhanc-
ties, as well as a potential to be used as a chemopre-
ing properties (6) and can promote cell invasiveness (7).
ventive agent in humans (17,18). It seems that mecha-
These abilities of galectin-3 are closely associated with
nisms underlying these effects involve suppression of
assets of tumorigenesis and metastasis (8–10) (e.g. ecto-
regulator(s) involved in activation of transcription fac-
pic expression or inhibition of galectin-3 has been shown
tors c-Jun and NF-kB (nuclear factor kappa B) (19,20).
either to increase or decrease tumorigenicity and meta-
According to the results of our previous studies both of
static proclivity of tumor cells, respectively (11–13)). Re-
these factors are involved in the regulation of the basal
cently, elevation of galectin-3 level has become a useful
expression of galectin-3 as well as of induced expression
diagnostic marker for breast carcinoma (14), chordoma
of galectin-3 caused by alkylating agent N-methyl-N'-nitro-
(15), thyroid carcinoma (16) and some other tumors.
N-nitrosoguanidine (MNNG) and ultraviolet C (UV-C)
* Corresponding author; Phone: +385 1 4818 757; Fax: +385 1 4856 201; E-mail: jdumic@pharma.hr

282
JERKA DUMI] et al.: Curcumin – Inhibitor of Galectin-3 Expression, Food Technol. Biotechnol. 40 (4) 281–287 (2002)
light (21). This is why both MNNG and UV-C light were
phosphatase were from Sigma (St Lewis, MO). Antibo-
used as model stressors in this study of the ability of
dies against c-Jun were from Oncogene Research
curcumin to inhibit the expression of galectin-3.
Products (Cambridge, MA).
Materials and Methods
Cell lines, cell culture, and stress procedures
We have recently shown that human glioblastoma
Materials
A1235 cells undergo prompt glycosylation changes after
All chemicals were of analytical grade and purcha-
exposure to different stressors (22,21) and all experiments
sed from Sigma (St. Louis, MO, USA), if not stated oth-
described here were performed on this cell-line. Cells
erwise. Nitrocellulose membranes were purchased from
were asynchronously grown (37 °C, 5 % CO2, relative
Millipore Corp. (Bedford, MA, USA), and heat-inactiva-
humidity 95 %) in the Dulbecco’s modified Eagle’s me-
ted fetal calf serum (FCS) from Life Technologies (Rock-
dium (Sigma, Cat. No. D5648) supplemented with 10 %
ville, MD, USA). Rat monoclonal antibody M3/38 (anti-
heat-inactivated FCS, 100 mg/mL streptomycin and 100
galectin-3) was kindly provided by Dr. I. Rosenberg
U/mL penicillin. Cells were subcultured twice a week
(Harvard Medical School, Boston, MA). MNNG, anti-
to maintain cultures in exponential growth.
mouse immunoglobulin G (IgG) labeled with alkaline
In all experiments cells were plated in tissue culture
phosphatase, and anti-rat IgG labeled with alkaline
dishes (60 ´ 15 mm) 24 h before treatment.
A
Control
UV (80 J/m 2)
curcumin (50 mM)
curcumin + UV
Gal-3
2 h
4 h
24 h
2 h
4 h
24 h
2 h
4 h
24 h
Time after exposure to UV light
B
2.5
2.0
**
area 1.5
**
1.0
Relative
*
*
0.5
0
2 h
4 h
24 h
2 h
4 h
24 h
2 h
4 h
24 h
Time after exposure to UV light
1
2
3
4
5
6
7
8
9
10
2
control cells
UV (80 J/m2 )
curcumin ( 50 mM)
curcumin (50 mM) + UV (80 J/m )
Fig. 1. Effects of curcumin on basal and UV-C-induced expression of galectin-3.
A1235 glioblastoma cells were pre-incubated for 1 h in medium with or without curcumin (50 mM), exposed or not to UV-C radiation (80
J/m2) and analyzed for galectin-3 (Gal-3) after 2, 4 and 24 h of further cultivation. Control cells were exposed neither to curcumin nor to
UV-C radiation.
A. Western blot analysis of galectin-3. Cellular proteins (10 mg per lane) were separated by 12 % SDS-PAGE, transferred to Immobilon-P
membranes and analyzed with antibodies against galectin-3.
B. The amount of galectin-3 was estimated by densitometric analysis and expressed as normalized values relative to the level of galec-
tin-3 in the control cells. Average values (± standard deviation (SD)) from two independent series of experiments (analyzed in duplica-
tes) are shown.
* - p < 0.05 comparing to the controls, ** - p < 0.05 compared to the corresponding curcumin-untreated cells

JERKA DUMI] et al.: Curcumin – Inhibitor of Galectin-3 Expression, Food Technol. Biotechnol. 40 (4) 281–287 (2002)
283
A
Control
UV (80 J/m 2)
curcumin (50 mM)
curcumin + UV
Jun
2 h
4 h
24 h
2 h
4 h
24 h
2 h
4 h
24 h
Time after exposure to UV light
B
88
77
66
55
area
44
Relative 33
**
22
**
**
*
11
*
00
2 h
4 h
24 h
2 h
4 h
24 h
2 h
4 h
24 h
Time after exposure to UV light
1
2
3
4
5
6
7
8
9
10
2
control cells
UV (80 J/m2 )
curcumin (50 mM)
curcumin (50 mM) + UV (80 J/m )
Fig. 2. Effects of curcumin on basal and UV-C-induced expression of c-Jun.
A1235 glioblastoma cells were pre-incubated for 1 h in medium with or without curcumin (50 mM), exposed or not to UV-C radiation (80
J/m2) and analyzed for c-Jun after 2, 4 and 24 h of further cultivation. Control cells were exposed neither to curcumin nor to UV-C radia-
tion.
A. Western blot analysis of c-Jun. Cellular proteins (10 mg per lane) were separated by 12 % SDS-PAGE, transferred to Immobilon-P
membranes and analyzed with antibodies against c-Jun.
B. The amount of c-Jun was estimated by densitometric analysis and expressed as normalized values relative to the level of c-Jun in the
control cells. Average values (±SD) from two independent series of experiments (analyzed in duplicates) are shown.
* - p < 0.05 compared to the controls, ** - p < 0.05 compared to the corresponding curcumin-untreated cells
To study the effects of UV-C light, cultivation me-
and fresh medium containing 50 mM curcumin (5 mL/
dium was removed and fresh medium (5 mL/culture)
culture) was added. In the control cell cultures fresh me-
was added. After 1 h the medium was removed and to
dium without curcumin was added. Cells were cultiva-
prevent the drying of the cells during irradiation, the
ted for 1 h before adding MNNG, or exposure to UV-C
amount of fresh medium, which corresponds to 1 mm
light, respectively.
high medium layer, was added (2.8 mL). Cells were ir-
Before analysis, cells were washed twice with ice-
radiated by Phillips UVT 15W/G15T T8 lamp with emis-
-cold 0.137 mol/L NaCl, 2.7 mmol/L KCl, 4.3 mmol/L
sion maximum at 254 nm. The intensity of irradiation
Na
was determined using BioBlock VLX-3W dosimeter.
2HPO4 × 7 H2O, 1.4 mmol/L KH2PO4, pH = 7.4 (PBS),
scraped and centrifuged at 600 ´ g for 10 min. The cell
Control cell cultures were kept at the room temperature
pellet was lyzed in 1 % Triton X-100 in 5 mM Tris/HCl,
during that time. After the exposure to the radiation
pH = 8.0, 15 mM NaCl, 2 % Na-azide, 2 mM phenyl-
(10–15 s), all cell cultures were supplemented with re-
methylsulfonyl fluoride (PMSF).
moved medium and returned into the incubator for the
next 2, 4 and 24 h.
For treatment with MNNG, cultivation medium was
Western-blot analysis
removed and fresh medium (5 mL/culture) was added.
Sodium dodecyl sulfate – polyacrylamide gel elec-
After 1 h, MNNG (5 mM) was added into medium. In
trophoresis (SDS-PAGE) was performed in 12 % poly-
the control cell, MNNG was omitted. Cells were culti-
acrylamide gels according to Laemmli (23). Separated
vated for the next 4 or 24 h under standard conditions.
proteins were transferred onto nitrocellulose membrane
To examine effects of curcumin on expression of
using semi-dry transfer technique. Membranes were
galectin-3 and c-Jun, cultivation medium was removed
blocked overnight with 3 % bovine serum albumin

284
JERKA DUMI] et al.: Curcumin – Inhibitor of Galectin-3 Expression, Food Technol. Biotechnol. 40 (4) 281–287 (2002)
A
Control
M
C
C + M
M
C
C + M
Gal-3
4 h
24 h
Time after adding of MNNG
B
1.8
1.6
1.4
a
**
r
e

1.2
a
1
*
*
0.8
Relative
0.6
0.4
0.2
0
4 h
24 h
Time after adding of MNNG
1
2
3
4
5
6
7
8
control cells
MNNG (5 mM)
curcumin (50 mM) curcumin (50 mM) + MNNG (5 mM)
Fig. 3. Effects of curcumin on basal and MNNG-induced expression of galectin-3.
A1235 glioblastoma cells were pre-incubated for 1 h in medium with or without curcumin (50 mM), MNNG was or was not added to the
medium (to the final concentration of 5 mM) and analyzed for galectin-3 (Gal-3) after 2, 4 and 24 h of further cultivation. Control cells
were exposed neither to curcumin nor to MNNG.
A. Western blot analysis of galectin-3. Cellular proteins (10 mg per lane) were separated by 12 % SDS-PAGE, transferred to Immobilon-P
membranes and analyzed with antibodies against galectin-3.
B. The amount of galectin-3 was estimated by densitometric analysis and expressed as normalized values relative to the level of galec-
tin-3 in the control cells. Average values (±SD) from two independent series of experiments (analyzed in duplicates) are shown.
* - p < 0.05 compared to the controls; ** - p < 0.05 compared to the corresponding curcumin-untreated cells
(BSA) in 0.05 mol/L Tris/HCl, pH = 7.5, 0.15 mol/L
Other procedures
NaCl (TBS). Galectin-3 was identified with anti-galec-
Protein concentrations in cell homogenates were de-
tin-3 antibodies (culture supernatant from hibridoma
termined by the method of Lowry (24).
M3/38) diluted 1:50 in 3 % BSA in TBS. c-Jun was iden-
tified with polyclonal rabbit anti-Jun antibodies (1:1000
in 3 % BSA in TBS). After incubation with alkaline phos-
phatase-labeled secondary antibody (goat anti-rat IgG
Results
diluted 1:2000 in TBS for galectin-3 and goat anti-rabbit
IgG diluted 1:1000 for c-Jun), proteins were visualized
with 0.02 g/L 5-bromo-4-chloro-3-indolyl phosphate
Effects of curcumin on basal
and 0.04 g/L nitro-blue tetrazolium in 0.05 mol/L Tris/
galectin-3 and c-Jun expression
HCl, pH = 9.5, 0.1 mol/L NaCl, 0.005 mol/L MgCl2.
Incubation of A1235 glioblastoma cells with curcu-
Membranes were incubated in dark until the color
min (50 mM) for 3 h did not result in statistically signifi-
developed (15 min on average) and digitalized with
cant change of galectin-3 level (Fig. 1, bar 5) or c-Jun
UMAX Astra 610P Scanner using 300 ´ 300 dpi optical
level (Fig. 2, bar 5). Additional 2 h-incubation resulted
resolution. Quantification of the bands corresponding to
in approximately 25 % decrease of galectin-3 level (Fig.
galectin-3 or c-Jun was performed by two-dimensional
1, bar 6; Fig. 3, bar 3) as well as 20 % decrease of c-Jun
analysis (spot densitometry) in National Institute of Health
level (Fig. 2, bar 6, Fig. 4, bar 3). After prolonged incu-
(NIH) software (»Tnimage Scientific Image Analysis
bation (25 h) galectin-3 level remained in the reached
Software«, by T.J. Nelson). Single factor ANOVA was
range (Fig. 1, bar 7; Fig. 3, bar 7) while c-Jun level fell
used to measure statistical significance. Difference of p
further, on approximately 65 % of the initial value (Fig.
< 0.05 was considered to be statistically significant.
2, bar 7; Fig. 4, bar 7).

JERKA DUMI] et al.: Curcumin – Inhibitor of Galectin-3 Expression, Food Technol. Biotechnol. 40 (4) 281–287 (2002)
285
A
Control
M
C
C + M
M
C
C + M
Jun
4 h
24 h
Time after adding of MNNG
B
2
1.8
1.6
a
1.4
r
e
a

1.2
**
1
*
0.8
* **
Relative
0.6
0.4
0.2
0
4 h
24 h
Time after adding of MNNG
1
2
3
4
5
6
7
8
control cells
MNNG (5 mM)
curcumin (50 mM) curcumin (50 mM) + MNNG (5 mM)
Fig. 4. Effects of curcumin on basal and MNNG-induced expression of c-Jun.
A1235 glioblastoma cells were pre-incubated for 1 h in medium with or without curcumin (50 mM), MNNG was or was not added to the
medium (to the final concentration of 5 mM) and analyzed for c-Jun after 2, 4 and 24 h of further cultivation. Control cells were exposed
neither to curcumin nor to MNNG.
A. Western blot analysis of c-Jun. Cellular proteins (10 mg per lane) were separated by 12 % SDS-PAGE, transferred to Immobilon-P
membranes and analyzed with antibodies against c-Jun.
B. The amount of c-Jun was estimated by densitometric analysis and expressed as normalized values relative to the level of c-Jun in the
control cells. Average values (±SD) from two independent series of experiments (analyzed in duplicates) are shown.
* - p < 0.05 compared to the controls; ** - p < 0.05 compared to the corresponding curcumin-untreated cells
Effects of curcumin on galectin-3 and c-Jun
8 vs. bar 2, bar 9 vs. bar 3, bar 10 vs. bar 4). Curcumin
expression in cells exposed to UV-C light
pre-treatment had the same effect on c-Jun expression.
Although c-Jun levels in curcumin-pretreated UV-C
Cell exposure to UV-C radiation of 80 J/m2 has in-
light exposed cells were 2.1, 1.5 and 1.2 times higher
duced expression of galectin-3. Two hours after irradia-
compared to the control after 2, 4 and 24 h, respectively
tion cell content of galectin-3 was approximately 60 %
(Fig. 2, bars 8, 9 and 10), they were remarkably reduced
higher compared to the initial values (Fig. 1, bar 2). Fur-
compared to the cells untreated with curcumin before
ther incubation was followed by additional elevation of
radiation by 43, 78 and 71 %, respectively (Fig. 2, bar 8
galectin-3 contents. After 4 and 24 h it reached levels
vs. bar 2, bar 9 vs. bar 3, bar 10 vs. bar 4).
that were 70 % and 80 % higher than the initial one (Fig.
1, bars 3 and 4). At the same time c-Jun levels (2, 4 and
24 h), were found to be increased compared to the con-
Effects of curcumin on galectin-3 and c-Jun
trols by 3.7, 6.7 and 4.2 times, respectively (Fig. 2, bars 2,
expression in cells exposed to MNNG
3 and 4).
Alkylating agent MNNG provoked elevation of ga-
Cell pre-exposure to curcumin (50 mM) for 1 h cau-
lectin-3 and c-Jun expression. Within first 4 h after addi-
sed considerable suppression of inducible effect of UV-C
tion of MNNG, galectin-3 amount increased for 54 %
radiation on galectin-3 expression. Galectin-3 levels in
compared to the controls (Fig. 3, bar 2) and stayed ele-
the cells treated with curcumin for 1 h before UV-C ex-
vated in the same range for the next 20 h of incubation
posure were still elevated compared to the controls by
(Fig. 3, bar 6). Intensity of c-Jun elevation was in the ap-
40, 38 and 26 % after 2, 4 and 24 h, respectively (Fig. 1,
proximately same range. Namely, after 4 h it reached
bars 8, 9 and 10), but after 4 and 24 h they were signifi-
the level that was 63 % higher than the initial one (Fig. 4,
cantly lower compared to the corresponding curcumin-
bar 2), and after 24 h it was slightly lower, but still 43 %
-untreated cells by 20 and 30 %, respectively (Fig. 1, bar
higher than in controls (Fig. 4, bar 6).

286
JERKA DUMI] et al.: Curcumin – Inhibitor of Galectin-3 Expression, Food Technol. Biotechnol. 40 (4) 281–287 (2002)
When cells were incubated with curcumin for 1 h
further reduction of galectin-3 level, the 75 % of the con-
before exposure to MNNG, inducible effect of MNNG
trol level, achieved after 5 h, was maintained. At the
on galectin-3 expression was slightly reduced after 4 h,
same time c-Jun level, after reduction of 20 % after 5 h
while after 24 h it was completely abolished. Namely, 4 h
continued to fall, and after 25 hours it reached 65 % of
after MNNG-exposure galectin-3 level in the curcumin-
the control value. In our earlier studies, Jun was recog-
-treated cells was 35 % higher compared to the control
nized as a regulator of the unstimulated expression of
(Fig. 3, bar 4), but after 24 h the significant difference
galectin-3, since specific inhibition of jun by antisense-
compared to the controls was not found (Fig. 3, bar 8).
-jun oligonucleotides significantly decreased basal levels
However, the tendency of curcumin to suppress induc-
of galectin-3, while specific inhibitor of NF-kB, carboxy-
ible effect of MNNG on galectin-3 expression noticed af-
benzyl-leucyl-leucyl-leucine vinyl sulfone (zL3-vs) did
ter 4 h of incubation (Fig. 3, bar 4 vs. bar 2) became sta-
not show this effect (21). Taken together, these facts sug-
tistically significant after 24 h of incubation (Fig. 3, bar 8
gest that curcumin affects galectin-3 expression in un-
vs. bar 6).
stimulated cell through transcription factor AP-1, but
Curcumin had even stronger effect on c-Jun expres-
that some other transcription factor could also be in-
sion. Already after 4 h there was no statistically signifi-
volved in this process, since the decrease of c-Jun level
cant difference between c-Jun level in the cells pretreat-
was more pronounced than the decrease of galectin-3.
ed with curcumin before MNNG exposure and the
In this work, we have also shown that curcumin in-
control level (Fig. 4, bar 4), while after 24 h c-Jun level
hibited the elevation of galectin-3 level induced by UV-
was reduced by 30 % compared to the control level (Fig.
-light radiation and alkylation agent MNNG. Curcumin
4, bar 8). In comparison with the corresponding curcu-
exerted more significant inhibitory effect when galec-
min untreated cells, cells treaded with curcumin for 1 h
tin-3 expression was induced with MNNG than with
before MNNG exposure contained approximately 50 %
UV-C light. Namely, inducible effect of MNNG was
lower c-Jun amount after 4 and 24 h (Fig. 4, bar 4 com-
abolished after 24 h, while UV-light-provoked stimula-
pared to bar 2, and bar 8 compared to bar 6).
tion was not fully eliminated. Similar result was obtai-
ned for c-Jun, but it is important to emphasize that UV-C
Discussion
radiation provoked huge elevation of c-Jun level (6.7
times higher level comparing to the controls was de-
The induction of galectin-3 expression is tightly re-
tected after 4 hours), while MNNG induced it mostly by
lated to tumorigenicity and malignancy of many kinds
60 % compared to the controls. Such a huge elevation of
of neoplasms (8–10) at both the protein and the mRNA
c-Jun level was not followed by a strong induction of
levels. Conversely, the inhibition of galectin-3 expression
galectin-3. In our previous studies it was noticed that
reduces tumor invasiveness and its metastatic potential
when c-Jun expression was significantly suppressed by
(11–13). Unraveling the interplay of the regulatory fac-
antisense-jun oligonucleotides, there was still some in-
tors involved in galectin-3 expression is an important is-
crease in galectin-3 after the exposure to UV-C light. It
sue (25–29) and biomedicine is highly interested in iden-
was also shown that UV light-induced expression of ga-
tifying likely inhibitors that restrain the process of ga-
lectin-3 related transcription factor NF-kB. Curcumin in-
lectin-3 expression.
hibits activation of both of these factors, hence uncom-
It was shown that in the range of 20 to 200 mM con-
pleted suppression of UV light-induced expression of
centrations curcumin, a natural, biologically active com-
galectin-3 achieved with curcumin suggests that some
pound extracted from turmeric, exhibits a broad spectrum
other factors could be involved in this regulatory mech-
of beneficial biological effects, e.g. anti-inflammatory,
anism. On the other hand, inducible effect of MNNG on
anti-mutagenic, anti-proliferative, and anti-cancerogenic
galectin-3 expression was abolished with curcumin
activities, without any toxic effect (30–32). Although the
treatment, so the central role in regulation of MNNG in-
complete genomic sequence of human galectin-3 gene
duced expression of galectin-3 could be attributed to the
(LGALS3) has been reported, the regulation of LGALS3
transcription factors c-Jun and NF-kB.
expression is still largely unknown (33). Analysis of the
The key findings of this study include: (i) curcumin
promotor region of LGALS3 revealed the existence of
(50 mM) was found to inhibit basal expression of galec-
multiple putative regulatory elements (e.g. Sp1, AP-1,
tin-3 and c-Jun in human glioblastoma A1235 cells. (ii)
NF-kB and cAMP response element binding factor
Cell pretreatment with curcumin (50 mM) for 1 h consid-
(CREB)). Recently we have shown that the cell exposure
erably reduced UV-C light-induced expression of galec-
to alkylating agent MNNG as well as UV light (l=254
tin-3 and c-Jun. (iii) Cell pretreatment with curcumin (50
nm) induce galectin-3 expression and that both trans-
mM) for 1 h completely abolished MNNG-induced ex-
cription factors, Jun (AP-1) and NF-kB take part in the
pression of galectin-3 and c-Jun. This study strongly sug-
regulatory mechanism of galectin-3 expression (21).
gests that curcumin is a potent inhibitor of galectin-3 ex-
These findings, as well as the fact that c-Jun and NF-kB
pression, and that it should be considered as a possible
activation can be inhibited by curcumin (18), encoura-
drug for treatment of pathological conditions character-
ged us to explore effects of curcumin on galectin-3 ex-
ised by elevated expression of galectin-3.
pression.
In this work we applied 50 mM concentration of
Acknowledgements
curcumin during 3, 5, and 25 h of incubation. Results
showed that it took 5 h of incubation in the presence of
This work was supported by grants #006320 and
curcumin until the decrease of galectin-3 level could be
#006320-2 from the Croatian Ministry of Science and
seen. Prolonged incubation of 25 hours did not result in
Technology.

JERKA DUMI] et al.: Curcumin – Inhibitor of Galectin-3 Expression, Food Technol. Biotechnol. 40 (4) 281–287 (2002)
287
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Kurkumin – sna`an inhibitor ekspresije galektina-3
Sa`etak
Galektin-3, lektin koji specifi~no prepoznaje b-galaktozidne strukture glikoproteina,
sudjeluje u razli~itim biolo{kim procesima. Patofiziolo{ka stanja i razli~iti uzro~nici stresa
bitno utje~u na ekspresiju galektina-3. Premda nisu poznati svi ~imbenici u regulacijskom
mehanizmu ekspresije galektina-3, pokazano je da u tom procesu sudjeluju transkripcijski
faktori AP-1 i NF-kB. Kurkumin, biolo{ki aktivan spoj izoliran iz biljnih podanaka vrste
Curcuma, interferira s djelovanjem obaju transkripcijskih faktora. Istra`ili smo u~inak kur-
kumina na ekspresiju galektina-3 u stanicama glioblastoma u bazalnim uvjetima te u stre-
su koji je bio izazvan izlaganjem stanica alkiliraju}em agensu N-metil-N'-nitro-N-nitrozo-
gvanidinu (MNNG) i ultraljubi~astom C (UV-C) svjetlu. Razina galektina-3 odre|ena je
tehnikom western-blot uz uporabu monoklonskih protutijela M3/38. Utvr|eno je da kur-
kumin smanjuje bazalnu razinu galektina-3 te da predinkubacija stanica kurkuminom jako
smanjuje inducibilni u~inak UV-C zra~enja, a potpuno uklanja induktivni u~inak alkilira-
ju}eg agensa MNNG. Rezultati nedvojbeno pokazuju da je kurkumin sna`an inhibitor eks-
presije galektina-3.

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