Biomedicine & Pharmacotherapy 67 (2013) 43–47

Original article

Resveratrol affects the cross talk between immune and colon cancer cells

Michael Bergman a,*,c, Gal Sahav Levin a,c, Hanna Bessler b,c, Meir Djaldetti b,c, Hertzel Salman a,c

a Department of Internal Medicine C, Rabin Medical Center, Hasharon Hospital, 7, Keren Kayemet Street, Petah-Tiqva, Israelb Laboratory for Immunology and Hematology Research, Rabin Medical Center, Hasharon Hospital, Petah-Tiqva, Israelc Sackler School of Medicine, Tel-Aviv University, Ramat-Aviv, Israel

A R T I C L E I N F O

Article history:

Received 21 October 2012

Accepted 23 October 2012

Keywords:

Resveratrol

Cytokines

Mononuclear cells

Colon cancer cells

A B S T R A C T

Resveratrol, a natural polyphenolic compound found mainly in grapes and their seeds, is gaining a

widespread appreciation as a therapeutic adjuvant in a variety of diseases including cancer prevention.

We examined the effect of resveratrol as a modulator of the immune dialog between peripheral blood

mononuclear cells and those from two human colon carcinoma lines, expressed by a possible alteration

of cytokine production. Resveratrol, incubated with non-stimulated mononuclear cells, caused a certain

reduction of IL-6, IL-1ra and IL-10, and a moderate increase of TNFa release. On the other hand,

resveratrol did not affect cytokine production by cancer cells from both lines. When resveratrol was

added to immune and cancer cells jointly, an altered dose-dependent decreased production of the

examined cytokines was obtained. These results favor the existence of a mechanism, additional to those

already described, that may explain the preventive effect of resveratrol on tumorigenesis.

� 2012 Elsevier Masson SAS. All rights reserved.

Available online at

www.sciencedirect.com

1. Introduction

The effect of nutrients on human health has been firmlyestablished. The majority of them poses several biological activities,such as inhibition of oxidative processes, acting as anti-inflamma-tory factors, and even exerting anti-cancer properties, observedmainly in vitro. Recently, following reports on its beneficial effect ona wide range of diseases thoroughly reviewed by Yadav et al. [1], theapplication of resveratrol as a therapeutic adjuvant gains a risinginterest. Studies have shown that the French Paradox could beexplained by moderate red wine consumption that induces a cardio-protective effect due to the angiogenic, antihypercholesterolemicand antidiabetic properties of resveratrol, a polyphenolic compoundpresented in the skin of red grapes and red wine [2–4]. The biologicaleffects of resveratrol occur through various systemic, cellular andeven sub-cellular pathways that have been reviewed in a number ofpapers [5–8]. Considering the close relationship between inflam-mation and cancer development [9], studies have been carried out toconnect the carcino-preventing effect of resveratrol with its anti-inflammatory activities. Apparently, the anti-inflammatory proper-ties of resveratrol occur by a variety of mechanisms, such asinhibited synthesis of pro-inflammatory mediators, inhibition ofimmune cells and suppression of cyclooxygenase-2 production [10].In addition, it has been shown that the inhibitory activity ofresveratrol on monocyte respiratory burst plays an important role in

* Corresponding author. Tel.: +972 3 9372306; fax: +972 3 9372432.

E-mail address: bermanm@clalit.org.il (M. Bergman).

0753-3322/$ – see front matter � 2012 Elsevier Masson SAS. All rights reserved.

http://dx.doi.org/10.1016/j.biopha.2012.10.008

inflammatory suppression [11,12]. Previous in vitro studies in ourlaboratory have implied that the inflammatory response linked withcolon cancer development may proceed through a cross talkbetween immune and cancer cells [13]. This process may bemodulated by a number of drugs, for example aspirin and statins[14,15], as well as by nutrients, such as caffeine and curcumin[16,17]. The purpose of the present study was to address thepossibility that resveratrol may affect the cross talk between humanperipheral blood mononuclear cells (PBMC) and those from twohuman colon cancer lines i.e. HT-29 and RKO. The results mayenlighten the way by which resveratrol affects cancer development.

2. Materials and methods

2.1. Cell preparation

PBMC were separated from venous blood obtained from adultblood bank donors by gradient centrifugation using Lymphoprep-1077 (Axis-Shield PoC AS, Oslo, Norway). The cells were washedtwice in phosphate buffered saline (PBS) and suspended in RPMI-1640 medium (Biological Industries, Beith Haemek, Israel)containing 1% penicillin, streptomycin and nystatin, 10% fetal calfserum (FCS), and designated as complete medium (CM).

2.2. Cell lines

HT-29 and RKO human colon cancer cell lines were obtainedfrom the American Type Cultural Collection (ATCC), Rockville, MD.The cells were maintained in CM containing MacCoy’s 5A

0 10 25 60 0 10 25 60

0

50

100

150HT-29 RK O

* *

* * ***

* *

Resve ratrol conce ntration , µM

XT

T t

es

t, %

of

co

ntr

ol

Fig. 1. Effect of resveratrol on the proliferation rate of HT-29 and RKO colon cancer

cells tested by XTT-proliferation test. 4 � 104 cells/well were incubated for 24 hours

with or without resveratrol at doses as indicated. At the end of incubation period

XTT test was performed. The results were calculated as percent of the control (cells

incubated without Resveratrol). Each column represents the mean of 7

experiments. Bars represent SEM. Asterisks represent statistically significant

difference from peripheral blood mononuclear cells incubated without resveratrol

(**P < 0.01, ***P < 0.001).

M. Bergman et al. / Biomedicine & Pharmacotherapy 67 (2013) 43–4744

(Biological Industries Co, Beth-Haemek, Israel), or modified eaglemedium (MEM- Biological Industries Co., Beth-Haemek, Israel)respectively, supplemented with 10% FCS, 2 mM L-glutamine andantibiotics (penicillin, streptomycin and nystatin-Biological In-dustries Co, Beth-Haemek, Israel). The cells were grown in T-75culture flasks (Nunc, Roskidle, Denmark) at 37 8C in a humidifiedatmosphere containing 5% CO2.

2.3. Resveratrol

Resveratrol (Sigma, Israel) was dissolved in dimethyl sulfoxide(DMSO- Sigma, Israel) at a concentration of 100 mM and kept at –20 8C. Further dilutions were prepared in DMSO before use.Resveratrol was added at final concentrations of 10, 25 and 60 mM.The concentration of DMSO in the cultures with and withoutreservatrol (controls) was 0.1%.

2.4. Effect of resveratrol on cytokine production by peripheral blood

mononuclear cells induced by cancer cells

0.5 ml of PBMC (4 � 106/ml of CM) was incubated with equalvolume of CM or with each type of cancer cells (4 � 105/ml of CM)suspended in the appropriate CM. Resveratrol was added at theonset of cultures at concentrations as described above. Thecultures were incubated for 24 hours at 37 8C in a humidifiedatmosphere containing 5% CO2. At the end of the incubation period,the cells were removed by centrifugation at 1500 rpm for10 minutes, the supernatants were collected and kept at –75 8Cuntil assayed for cytokine content.

2.5. Effect of resveratrol on the secretion of colon cancer cells’

conditioned medium

Colon cancer cells suspended at 4 � 105/ml in the appropriateCM were incubated for 24 hours at 37 8C in a humidifiedatmosphere containing 5% CO2 with or without resveratrol atconcentrations as indicated. At the end of the incubation period,the cells were removed by centrifugation at 1500 rpm for10 minutes and the conditioned media were assayed for theireffect on cytokine secretion by PBMC as follows: 0.5 ml of PBMC(4 � 106/ml of CM) were incubated for 24 hours at 37 8C with equalvolume of the various conditioned media. At the end of theincubation period, the cells were removed by centrifugation at1500 rpm for 10 minutes, the supernatants were collected andkept at –75 8C until assayed for cytokine content.

2.6. Effect of resveratrol on malignant cell proliferation (XTT test)

0.1 ml aliquots (4 � 105/ml) of HT-29 or RKO cells suspended inCM were placed in each one of 96-well flat-bottomed plates (Nunc,Roskidle, Denmark). Resveratrol was added at the onset of theculture at final concentrations of 10, 25 and 60 mM. The cultures,set up in triplicates, were incubated for 24 hours at 37 8C in ahumidified atmosphere supplemented with 5% CO2. At the end ofthe incubation period XTT test (Biological Industries, CO Beth-Haemek, Israel) was performed according to the instructionsprovided by the manufacturer.

2.7. Cytokine content in the supernatants

The concentration of the following cytokines: TNFa, IL-1b, IL-1ra, IL-6, IL-10 and IFNg in the supernatants was tested using ELISAkits specific for human cytokines (Biosource International,Camarillo, CA) as detailed in the guide-line provided by themanufacturer. The detection levels of these kits were: 15 pg/ml forIL-6, and 30 pg/ml for the others.

2.8. Statistics

Data was analyzed using ANOVA with repeated measures foreach cytokine, and paired t-test to compare the difference betweenthe levels of cytokine obtained with or without various concen-trations of resveratrol. P values of < 0.05 were considered asstatistically significant. The results are expressed as mean � SEM.

3. Results

3.1. Effect of resveratrol on malignant cell proliferation

A dose-dependent inhibition of the proliferation of both HT-29and RKO cell lines was found following 24 h of incubation withincreasing doses of resveratrol (F3,35 = 3.4, P = 0.034 andF3,35 = 7.12, P = 0.00136, respectively) (Fig. 1). At 10 mM ofresveratrol, the proliferation of both cell lines was not significantlyaffected. At resveratrol concentration of 25 and 60 mM, theproliferation of HT-29 cells was reduced by 18 and 27%,respectively (P = 0.008 and P = 0.002, respectively) and that ofRKO cells by 17 and 17% respectively (P = 0.0037and P = 0.0008,respectively).

3.2. Effect of resveratrol on cytokine production by cancer cells

Incubation of malignant cells with resveratrol at doses asindicated had no effect on the production of any of the cytokinesexamined in this study. Conditioned media from HT-29 or RKOcells cultured for 24 hours did not contain detectable levels of theabove- tested cytokines.

3.3. Effect of resveratrol on cytokine production by peripheral blood

mononuclear cells

The amount of cytokines secreted by non-stimulated PBMC wassignificantly lower than that produced by PBMC incubated for24 hours with cancer cells (Table 1). Incubation of PBMC withresveratrol caused a significant inhibition of the secretion of IL-6(F3,23 = 10.75, P = 0.0005) and IL-10 (F3,23 = 6.97, P = 0.0036),whereas that of TNFa was slightly stimulated (F3,23 = 4.13,

Table 1Effect of resveratrol on cytokine production by peripheral blood mononuclear cells (PBMC).

PBMC only PBMC + Resveratrol ANOVA

0 10 mM 25 mM 60 mM P value

TNFa, pg/ml 38 � 7 71 � 16 51 � 11 28 � 10 0.025

IFNg, ng/ml 0.15 � 0.03 0.16 � 0.03 0.14 � 0.02 0.13 � 0.02 0.085

IL-1b, ng/ml 1.63 � 0.14 1.73 � 0.23 1.59 � 0.16 1.31 � 0.15 0.056

IL-6, ng/ml 13.5 � 1.6 13.1 � 1.5 12.3 � 1.4* 10.5 � 1.9*** 0.0005

IL-1ra, pg/ml 238 � 35 257 � 38 232 � 31 200 � 17 0.086

IL-10, pg/ml 183 � 9 192 � 25 146 � 4** 122 � 4*** 0.0036

PBMC: peripheral blood mononuclear cells; 2 � 106 PBMC were incubated for 24 hours with or without resveratrol at doses as indicated. The cytokine levels in the

supernatants were tested by ELISA. The results of six experiments are expressed as mean � SEM. Asterisks represent statistically significant difference from PBMC incubated

without resveratrol* P < 0.01.** P < 0.01.*** P < 0.001.

M. Bergman et al. / Biomedicine & Pharmacotherapy 67 (2013) 43–47 45

P = 0.025). The production of IL-1b, IFNg and IL-1ra was notaffected by addition of resveratrol at the doses tested (F3,23 = 3.15,P = 0.056, F3,23 = 2.67, P = 0.085, and F3,23 = 2.65, P = 0.086,respectively).

3.4. Effect of resveratrol on pro-inflammatory cytokine production

3.4.1. TNFaTNFa secretion by PBMC induced by both HT-29 and RKO cells

was affected by incubation with increasing doses of resveratrol(F3;31 = 6.01, P < 0.0039 and F3;35 = 18.2, P < 0.0001, respective-ly) (Fig. 2A). At resveratrol concentration of 60 mM, TNFaproduction induced by both cell lines was reduced by 60%(P < 0.005). At lower concentrations of resveratrol, the secretionof TNFa induced by HT-29 was not affected whereas theproduction of TNFa by PBMC-induced by RKO cells was increasedby 25% (P < 0.001) at drug concentration of 10 mM.

Fig. 2. Effect of resveratrol on pro-inflammatory cytokine secretion by peripheral blood

conditioned media (Sups) derived from similar numbers of the same cells. 2 � 106 PBMC

resveratrol at doses as indicated, or with sups from cancer cells (2 � 105/ml) incubated fo

IL-1b (B), IL-6 (C) and IFN (D) levels in the supernatants was tested by ELISA. Each col

represent statistically significant difference from PBMC incubated without resveratrol

Conditioned media derived from HT-29 cells incubated with thethree doses of resveratrol had no effect on TNFa production byPBMC (F3;27 = 2.16, P = 0.128). However, conditioned mediaderived from RKO cells incubated with increasing doses of thedrug caused a dose-dependent stimulation of TNFa secretion byPBMC (F3;27 = 10.09, P < 0.001). Conditioned media derived fromRKO cells incubated with 25 or 60 mM of resveratrol caused 165and 265% increase in TNFa secretion by PBMC, respectively(P = 0.023, and P = 0.016, respectively).

3.4.2. IL-1bResveratrol affected the production of IL-1b by PBMC induced

by HT-29 or RKO cells (F3;31 = 7.82, P = 0.001 and F3;31 = 12.69,P < 0.001, respectively), however in different ways (Fig 2B). Atresveratrol concentrations of 10, and 25 mM, the secretion of IL-1binduced by HT-29 cells was slightly enhanced by 19% and 6%respectively (P = 0.05, and P = 0.064, respectively), whereas at

mononuclear cells (PBMC) induced by colon cancer cells (HT-29 and RKO) or their

were incubated for 24 hours with 2 � 105 cancer cells in the absence or presence of

r 24 hours with or without resveratrol at concentrations as indicated. The TNFa (A),

umn represents the mean of at least 6 experiments. Bars represent SEM. Asterisks

(*P < 0.05, **P < 0.01, ***P < 0.001).

PBM

C 0 10 25 60 0 10 25 60

0

1

2

3

4Cell -to-cell

HT-29 RKO

***

Sup ernatant

**

****

IL-1

0,

ng

/ml

PBM

C 0 10 25 60 0 10 25 60

0.0

0.5

1.0

1.5

2.0

2.5 HT-29 RKO

*

***

***

***

*

*

*

Resve ratrol concen tration , µM

Resve ratrol concen tration , µM

IL-1

ra,

ng

/ml

A

B

Fig. 3. Effect of resveratrol on anti-inflammatory cytokine secretion by peripheral

blood mononuclear cells (PBMC) induced by colon cancer cells (HT-29 and RKO) or

by their conditioned media (Sups) derived from similar numbers of the same cells.

2 � 106 PBMC were incubated for 24 hours with 2 � 105 cancer cells in the absence

or presence of resveratrol at doses as indicated, or with sups obtained from cancer

cells (2 � 105/ml) incubated for 24 hours with or without resveratrol at

concentrations as indicated. The IL-1ra (A) and IL-10 (B) levels in the

supernatants were tested by ELISA. Each column represents the mean of at least

6 experiments, Bars represent SEM. Asterisks represent statistically significant

difference from PBMC incubated without resveratrol (*P < 0.05, **P < 0.01,***P < 0.001).

M. Bergman et al. / Biomedicine & Pharmacotherapy 67 (2013) 43–4746

60 mM resveratrol caused 27% reduction in its secretion(P = 0.064). As for RKO-induced IL-1b production by PBMC,resveratrol at 10, 25 and 60 mM caused a reduced secretion ofIL-1b by PBMC by 8.5, 7 and 25%, respectively (P = 0.02, P = 0.12and P = 0.004, respectively).

Conditioned media derived from HT-29 cells incubated with orwithout resveratrol, added to PBMC induced a similar IL-1bproduction (F3;27 = 0.6, P = 0.623). However, conditioned mediaderived from RKO cells incubated with resveratrol caused a dose-dependent stimulation of IL-1b secretion (F3;27 = 5.23, P = 0.008,respectively). Twenty-five and 60 mM of resveratrol increased theproduction of IL-1b by 30 and 60%, respectively (P < 0.05).

3.4.3. IL-6

HT-29-induced IL-6 secretion was affected by incubation withresveratrol (F3;31 = 5.18, P = 0.0077) (Fig. 2C). At 10 mM, thesecretion of IL-6 was enhanced by 18% (P = 0.04) whereas at60 mM, the reduction was enhanced by 26% (P = 0.089). RKO-induced IL-6 production by PBMC was not affected by addition ofresveratrol at the three doses tested (F3;31 = 1.43, P = 0.262).

Conditioned media derived from cancer cells from the two linesincubated with or without resveratrol did not affect IL-6production differently (F3;27 = 1.23, P = 0.327 for HT-29 cellsand F3;23 = 1.37, P = 0.286 for RKO cells).

3.4.4. IFNgA dose-dependent inhibited secretion of IFNg by PBMC induced

by both HT-29 and RKO cells was observed following incubationwith resveratrol (F3;31 = 6.59, P = 0.0025, and F3;31 = 12.09,P = 0.00014, respectively) (Fig. 2D). At 60 mM of resveratrol, thesecretion of IFNg by PBMC induced by HT-29 was reduced by 42%(P = 0.012) and that induced by RKO cells was suppressed by 80%(P = 0.0049). At lower concentrations of the drug, the reducedsecretion of IFNg did not reach statistically significance. Condi-tioned medium from both cell lines incubated with or withoutresveratrol had no effect on IFNg production by PBMC (data notshown).

3.5. Effect of resveratrol on anti-inflammatory cytokine production

3.5.1. IL-1ra

Increasing doses of resveratrol caused a dose-dependentreduction of IL-1ra secretion by PBMC induced by HT-29(F3;31 = 5.59, P = 0.005), but had no effect on IL-1ra secretioninduced by RKO cells (F3;31 = 2.14, P = 0.125) (Fig. 3A). At 10, 25and 60 mM, HT-29-induced IL-1ra production was reduced by 7,11.5 and 42%, respectively (P = 0.43, P = 0.38 and P = 0.022,respectively). Conditioned media collected from HT-29 cellsincubated with or without resveratrol exerted a similar effect onIL-1ra secretion (F3;27 = 2.19, P = 0.124) whereas that obtainedfrom RKO cells caused a slightly reduced secretion of this cytokine(F3;27 = 4.0, P = 0.024).

3.5.2. IL-10

Resveratrol caused a dose-dependent decrease in IL-10 secre-tion by PBMC induced by cancer cells from both lines (F3;31 = 5.52P < 0.006, and F3;31 = 5.96, P = 0.004 for HT-29 and RKO cells,respectively) (Fig. 3B). At doses of 10, 25 and 60 mM, resveratrolinhibited IL-10 synthesis by PBMC induced by RKO cells by 10, 13and 35% respectively (P = 0.43, P = 0.018 and P = 0.015, respective-ly), whereas that induced by HT-29 cells was lowered only by60 mM (38% reduction, P = 0.019). Conditioned media derivedfrom HT-29 cells incubated with or without the three dosesof resveratrol, did not affect differently IL-10 secretion by PBMC(F3;27 = 1.18, P = 0.345). Conditioned media from RKO cellsincubated with different doses of resveratrol exerted a

dose-dependent inhibition of IL-10 release by PBMC(F3;27 = 10.63, P < 0.0003). Conditioned medium from RKO cellsincubated with 25 and 60 mM resveratrol reduced IL-10 secretionby PBMC by 6 (P = 0.42) and 28% (P = 0.0005), respectively.

4. Discussion

Normal activity of PBMC and their subsets is imperative forevolution of inflammatory responses. Stimulated by antigens, forexample lipopolysaccharide (LPS), one of their main functions is toproduce pro- and anti-inflammatory cytokines, thus playing animportant role in the initiation and progress of infection andinflammation [18]. Regarding the close relationship betweeninflammation and tumorigenesis, studies have shown that tumorcells are able to exert an immunomodulatory effect on LPSstimulated CD14+/Cd16+ monocyte subpopulations proddingthem to produce higher levels of TNFa and IL-10 and lower levelof IL-12, compared with LPS stimulated cells only [19]. Moreover,in vitro experiments in our laboratory have shown that tumor cellsare able to conduct an immunomodulatory dialog even with non-stimulated PBMC, inducing them to produce various cytokines.However, one must keep in mind that PBMC enfold a larger pool ofimmune cells in addition to CD14+/Cd16+ cells. The data herebypresented indicate that non-stimulated PBMC were able to

M. Bergman et al. / Biomedicine & Pharmacotherapy 67 (2013) 43–47 47

produce certain amount of both pro- and anti-inflammatorycytokines, whereas carcinoma cells alone did not producedetectable amounts of cytokines at all. On the other hand,incubation of PBMC with cells of the two colon carcinoma linesbrought to an increased cytokine production, indicating that thiscell-to-cell interaction serves as a trigger for cytokine generation. Itis plausible that in cases with colon carcinoma, this paradigm maylead to an increased production of pro-inflammatory cytokines bymononuclear cells, promoting a chronic inflammatory processwith successive tumor development. In the present work, additionof resveratrol to non-stimulated PBMC caused a reduced release ofIL-1b, IL-6 and IL-10 while the production of TNFa was increasedby lower doses and decreased by the higher one. This finding is ofinterest since resveratrol acted on non-activated PBMC, whileRichard et al. [20] could obtain increased expression of IL-6 and IL-8 upon applying resveratrol on stimulated cells only. However, itshould be emphasized that in distinction from our experiments,their study was carried out with suspensions containing bothmonocytes and polymorphonuclear cells. The dual dose-depen-dent pattern of TNFa production by PBMC incubated withresveratrol is important since TNFa acts not only as a pro-inflammatory cytokine but possesses antitumor activity [21], acharacteristic that may explain its dual effect on both inflamma-tion and tumorigenesis. While resveratrol exerted a statisticallysignificant effect on PBMC stimulating the production of both pro-inflammatory (TNFa and IL-6) and anti-inflammatory (IL-10)cytokines by PBMC/tumor cells interaction, it did not affect thegeneration of any of the cytokines examined by cancer cells of bothlines. Our results demonstrate that resveratrol acts as an inhibitorof human cancer cell proliferation, as it has been shown in otherstudies [22–24]. Moreover, it is able to modulate the inflammatorycytokine production initiated by an immune dialog between PBMCand cancer cells. In our previous study [13], we have demonstratedthat the amount of inflammatory cytokines released by PBMC/carcinoma interaction is directly proportional to the number of themalignant cells, resulting in dysregulation of cytokine equilibrium.Since resveratrol caused an inhibition of malignant cell prolifera-tion, one would expect that the secretion of pro-inflammatorycytokines would be suppressed. However, while the production ofmost of the cytokines was reduced at higher resveratrol doses, thatof IL-6 was not affected, and at low dose (10 mM) a significantstimulation of IL-1b, IL-6 and TNFa production was observed. Thepossible relationship between reduced colon cancer cell prolifera-tion caused by resveratrol and the lower secretion of cytokinescannot be ruled out.

In short, in addition to its capacity to modulate an array ofimmune cell functions, resveratrol is gaining an increasedreputation as a potential inhibitor of tumor development [25–27]. The present study, indicating that resveratrol may shift thebalance of the immune equilibrium between PBMC and tumorcells, forwards an additional pathway for explaining its anti-cancereffect.

Disclosure of interest

The authors declare that they have no conflicts of interestconcerning this article.

References

[1] Yadav M, Jain S, Bhardwaj A, Nagpal R, Puniya M, Tomar R, et al. Biological andmedicinal properties of grapes and their bioactive constituents: an update. JMed Food 2009;12:473–84.

[2] Kaur G, Roberti M, Raul F, Pendurthi UR. Suppression of human monocytetissue factor induction by red wine phenolic and synthetic derivatives ofresveratrol. Thromb Res 2007;119:247–56.

[3] Penumathsa SV, Thirunavukkarasu M, Koneru S, Juihasz B, Zhan L, Pant R, et al.Statin and resveratrol in combination induces cardioprotection against myo-cardial infarction in hypercholesterolemic rat. J Mol Cell Cardiol 2007;42:508–16.

[4] Penumathsa SV, Maulik N. Resveratrol: a promising agent in promotingcardioprotection against coronary heart disease. Can J Physiol Pharmacol2009;87:275–86.

[5] Szabo G. A glass of red wine to improve mitochondrial biogenesis? Novelmechanisms of resveratrol. Am J Physiol Heart Circ Physiol 2009;297:H8–9.

[6] Csiszar A, Labinskyy N, Pinto JT, Ballabh P, Zhang H, Losonczy G, et al.Resveratrol induces mitochondrial biogenesis in endothelial cells. Am J PhysiolHeart Circ Physiol 2009;297:H13–20.

[7] Marques FZ, Markus MA, Morris BJ. Resveratrol: Cellular actions of a potentnatural chemical that confers a diversity of health benefits. Int J Biochem CellBiol 2009;41:2125–8.

[8] Harikumar KB, Aggarwal BB. Resveratrol. A multitargeted agent for age-associated chronic diseases. Cell Cycle 2008;7:1020–35.

[9] Grivennikov SI, Karin M. Inflammatory cytokines and cancer: tumor necrosisfactor and interleukin 6 take the stage. Ann Rheum Dis 2011;70(Suppl 1):104–8.

[10] de la Lastra CA, Villegas I. Resveratrol is an anti-inflammatory and anti-agingagent: mechanisms and clinical implications. Mol Nutr Food Res 2005;49:405–30.

[11] Poolman TM, Ng LL, Manson MM. Inhibition of the respiratory burst byresveratrol in human monocytes: correlation with inhibition of PI3 K signal-ing. Free Radic Biol Med 2005;39:118–32.

[12] de la Lastra CA, Villegas I. Resveratrol as an antioxidant and pro-oxidant agent:mechanisms and clinical implications. Biochem Soc Trans 2007;35:1156–60.

[13] Bessler H, Djaldetti M. Role of the equilibrium between colon cancer andmononuclear cells in cytokine production. Biomed Pharmacother 2010;64:707–11.

[14] Bergman M, Djaldetti M, Salman H, Bessler H. Inflammation and colorectalcancer: does aspirin affect the interaction between cancer and immune cells?Inflammation 2011;34:22–8.

[15] Bergman M, Salman H, Djaldetti M, Bessler H. Statins as modulators of coloncancer cells induced cytokine secretion by human PBMC. Vasc Pharmacol2011;54:88–92.

[16] Bessler H, Salman H, Bergman M, Djaldetti M. Caffeine alters cytokine secre-tion by PBMC induced by colon cancer cells. Cancer Investig 2011;30:87–91.

[17] Bessler H, Djaldetti M. Curcumin affects the cross-talk between immunocytesand colon carcinoma cells. Recent Res Devel Nutr 2012;8:81–93.

[18] Ziegler-Heitbrock L. The CD14+ CD16+ blood monocytes: their role in infectionand inflammation. J Leukoc Biol 2007;81:584–92.

[19] Szaffarska A, Baj-Krzyworzeka M, Siedlar M, Weglarczyk K, Ruggiero I, Hajto B,et al. Antitumor response of CD14+/CD16+ monocyte subpopulation. ExpHematol 2004;32:748–55.

[20] Richard N, Porath D, Radspieler A, Schwager J. Effects of resveratrol, picea-tannol, tri-acetoxystilbene, and genistein on the inflammatory response ofhuman peripheral blood leucocytes. Mol Nutr Food Res 2005;49:431–42.

[21] Drutskaya MS, Efimov GA, Kruglov AA, Kuprash DV, Nedospasov SA. Tumornecrosis factor, lymphotoxin and cancer. IUBMB Life 2010;62:283–9.

[22] Li T, Fan GX, Wang W, Li T, Yuan YK. Resveratrol induces apoptosis, influencesIL-6 and exerts immunomodulatory effect on mouse lymphocytic leukemiaboth in vitro and in vivo. Int Immunopharmacol 2007;7:1221–31.

[23] Juan ME, Alfaras A, Pianas JM. Colorectal cancer chemoprevention by transre-servatrol. Pharmacol Res 2012;65:584–91.

[24] Vanamala J, Reddivari L, Radhaksishnan S, Tarver C. Resveratrol suppressesIGF-1 induced human colon cancer cell proliferation and elevates apoptosis viasuppression of IGF-1R/Wnt and activation of p53 signaling pathways. BMCCancer 2010;10:238–51.

[25] Shankar S, Singh G, Srivastava RK. Chemoprevention by resveratrol: molecularmechanisms and therapeutic potential. Front Biosci 2007;12:4839–54.

[26] Udenigwe CC, Ramprasath VR, Aluko RE, Jones PJH. Potential of resveratrol inanticancer and anti-inflammatory therapy. Nutr Rev 2008;55:445–54.

[27] Kaur M, Agarwal C, Agarwal R. Anticancer and cancer chemopreventivepotential of grape seed extract and other grape-based products. J Nutr2009;139:1806S–12S.