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Review

Resveratrol: Challenges in Translation to the Clinic — A CriticalDiscussion

Lalita Subramanian1,2, Sherry Youssef1, Saswati Bhattacharya1, Jason Kenealey1,3,Arthur S. Polans1,2,3,4, and Paul R. van Ginkel1

AbstractLow cancer survival rates and the serious side effects often associated with current chemotherapeutics

highlight the need for new and effective nontoxic anticancer agents. Since 1997 when Jang and colleagues

first described resveratrol’s ability to inhibit carcinogenesis, it has consistently proven effective at tumor

inhibition in diverse human cancer models. This finding has raised the hope that resveratrol would pioneer

a novel class of nontoxic chemotherapeutics. As a consequence of initial basic and preclinical studies,

resveratrol is now being extensively promoted in the unregulated nutraceutical sector. However, some

fundamental aspects of resveratrol’s action need to be understood before it can be developed into a

clinically viable anticancer drug. These areas pertain to the key mechanism(s) by which resveratrol

potentiates its antitumor effects. Current research suggests that these mechanisms might be through novel

pathways, requiring an understanding of cellular uptake, sentinel targets, and in vivo biological networks.

The metabolism of resveratrol and its bioavailablity also warrant further consideration in light of recent

in vitro and in vivo studies. Finally, we need to appreciate the sorts of information about resveratrol that may

translate between different disease entities. We present a critical discussion of these issues and suggest

important experiments that could pave the way to the successful translation of resveratrol to the clinic.

Clin Cancer Res; 16(24); 5942–8. �2010 AACR.

Now that 60 Minutes, Barbara Walters, National PublicRadio, Dr. Oz, and a myriad of columnists at the New YorkTimes and other newspapers have offered their commentsand recommendations about the health benefits of resver-atrol, including its potential anticancer properties, perhapsit is time for scientists to ask a few essential questions aboutthe popular product before considering a further pitch tothe public.

In 1997, Jang and colleagues published their seminalreport on resveratrol’s ability to inhibit the three majorstages of carcinogenesis (1). Since then, there has been anexplosion of literature about resveratrol’s effects on cancer.Some fundamental aspects of our understanding of resver-atrol, however, are yet to be resolved. These gaps in ourknowledge must be addressed before the full potential ofthis nontoxic compound as an anticancer drug can berealized.

Cellular Uptake

What evidence do we have that resveratrol actuallyenters cancer cells?

A critical first step to delineate the mechanisms of drugaction is to determine whether resveratrol mediates itseffects by cell surface receptors or intracellular targets intumor and other cells. Both intra- and extracellular resver-atrol targets have been proposed (see below, "Targets: Fastand Slow"). However, a direct-binding partner has yet to beconvincingly established. Elucidation of whether resvera-trol enters cells or acts at the cell surface will shed somelight on the targets on which it acts.

Resveratrol is a hydrophobic compound and, currently,the only compelling evidence for entry of resveratrol intocells comes from highly specialized cell types: intestinaland liver cells. When a monolayer of colonic Caco-2 cellswas treated in vitro with resveratrol, a saturating dose-dependent apical to basolateral transport was observed(2, 3). In another study, the human hepatoblastoma cellline HepG2 and human hepatocytes were used to compareresveratrol transport in normal and tumor cells (4). Resver-atrol uptake was detected by its intrinsic fluorescenceproperties as well as by using radiolabeled drug. Concen-tration and temperature dependence of uptake were exam-ined, and results indicated that both passive diffusion andcarrier-mediated transport might be involved.

The above cell types, however, have specialized uptakemechanisms. The intestinal epithelium is adapted for

Authors' Affiliations: 1Department of Ophthalmology and VisualSciences, 2Eye Research Institute; 3Department of Biomolecular Chem-istry, and 4Carbone Cancer Center, University of Wisconsin, Madison,Wisconsin

Corresponding Author: Paul van Ginkel, Department of Ophthalmologyand Visual Sciences, University of Wisconsin School of Medicine andPublic Health, Room K6/454 Clinical Sciences Center, 600 HighlandAvenue, Madison, WI 53792. Phone: 608-265-5409; Fax: 608-265-6021; E-mail: [email protected]

doi: 10.1158/1078-0432.CCR-10-1486

�2010 American Association for Cancer Research.

ClinicalCancer

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absorption of nutrients from food. A study of drug trans-port using a Caco-2monolayer may indicate transepithelialtransport of a drug, but does not necessarily imply thatother cells have similar uptake mechanisms. Similarly, oneof the main functions of hepatocytes is to extract protein-bound drugs and metabolites from circulation. Conse-quently, hepatocytes are equipped with many differenttransport proteins in the basolateral membrane. The onlyother evidence of possible resveratrol uptake has beenshown indirectly through sulfotransferase 1A1 activityassociated with two breast cancer cell lines, resulting inthe conversion of exogenously added resveratrol to resver-atrol 3-O-sulfate (5). These studies have not been replicatedin other tumor cells.If resveratrol cannot be shown to efficiently enter all

tumor cells, it may bemore likely that its antitumor effect isthe result of binding to an extracellular target. Therefore,further studies are necessary to directly address cellularuptake in cancer cells. New technology, such as two-photon microscopy, may be particularly appropriate fordetecting the real-time uptake of unmodified resveratroland its metabolites by taking advantage of their intrinsicfluorescence under conditions of minimal photo-bleach-ing. Results of such studies will indicate the validity ofintracellular targets identified thus far and aid in theidentification of new sentinel targets.

Targets: Fast and Slow

What are the direct targets of resveratrol, and can wedifferentiate binding events from indirect effects ofresveratrol?The wealth of resveratrol literature is, in part, a result of

the myriad pathways and molecules that are altered inresponse to resveratrol treatment (reviewed in ref. 6). Someof these effectsmay be cell-type specific, tissue-type specific,or dependent on whether a cell is transformed. Through

parametric analysis of gene set enrichment (PAGE analy-sis), resveratrol has been found to cause a significantalteration in 127 pathways (7). Certain effects of drugtreatment are measurable in a matter of seconds (8),whereas others only become apparent after hours or evendays. This finding, then, brings up the question of which ofthese effects is a direct result of resveratrol bindingand which are downstream effects of the initial targetinteraction.

Several molecular targets have been postulated as med-iators of resveratrol’s anticancer effects (7). Approachessuch as a phage-display screen using biotinylated resvera-trol identified several candidates (9). However, the directbinding of resveratrol to these targets in a cellular contexthas yet to be convincingly shown.

An example of a direct-binding target identified throughcolumn chromatography is quinone reductase 2 (10). Thebinding constant for NQO2 was in a physiologically rele-vant range. However, not all tumor cells have measurablelevels of this protein, despite having similar sensitivity tothe drug (11). Therefore, binding to this proteinmay not bepart of a general mechanism for resveratrol antitumoraction.

Other targets reported include DNA polymerase a, PKC,PKD, COX2, ribonucleotide reductase, and F0/F1 ATPase/ATP synthase, on the basis of alterations of enzymaticactivity. Only F0/F1 ATPase/ATP synthase, however, hasbeen shown to interact directly with resveratrol. An addi-tional complication in identifying and validating suchtargets is that binding and enzymatic experiments in vitroor in cells in culture may not reflect in vivo conditions.Because levels of unmetabolized resveratrol in vivo do notexceed the low micromolar range (11), it is possible thatsome observed target binding and modulations of path-ways are only induced because of exposure of isolatedenzymes or cells to constant and high drug concentrationsin vitro.

A more recent group of potential targets, the sirtuins, hasbeen the subject of intense research as mediators ofresveratrol’s life-extending activity as well as antitumoractivity. The direct binding of resveratrol to sirtuins hasnot been shown. Early work showing direct resveratrolactivation of purified SIRT1 using a fluorescently modifiedsubstrate for sirtuin was shown to be an artifact of the assay(12). Additionally, the effects of resveratrol on sirtuins incells seem to be measurable only after several hours. Thisfinding is suggestive of downstream or indirect effects, ifany at all. More recent studies also conflict with the originalobservation that resveratrol has an effect on Sir2 activity invivo (13), or an effect on the lifespan of yeast (13), Dro-sophila, or Caenorhabditis elegans (14).

Extracellular receptors for resveratrol such as insulingrowth factor receptor (IGFR) and integrin aVb3 have beenproposed. The IGFR study compared downstream activityusing known ligands (15). No direct binding was shown inthis study. The aVb3 study relied on competition assaysusing a known ligand and binding studies usingmembranefractions (16). Radiolabeled resveratrol was used to suggest

Translational Relevance

Side effects from chemotherapy highlight the need foreffective nontoxic anticancer agents. Resveratrol hasconsistently proven effective at tumor inhibition indiverse human cancer models, while remaining non-toxic. This finding suggests that resveratrol could pio-neer a novel class of chemotherapeutics. It is alreadybeing extensively promoted in the unregulated nutra-ceutical sector, despite recent controversies about targetinteraction, bioavailability, and other essential matters.However, fundamental aspects of resveratrol actionneed to be understood before it can be developed intoa clinically viable anticancer drug. We present a short,but critical, discussion of these issues and suggestimportant experiments that could pave the way to thesuccessful translation of resveratrol to the clinic.

Resveratrol: Challenges in Translation to the Clinic

www.aacrjournals.org Clin Cancer Res; 16(24) December 15, 2010 5943




direct binding to the b3 subunit of the integrin. However,given its properties, resveratrol is likely to associate withmembranes readily, making it difficult to assess the speci-ficity of these binding studies. Even though further work isneeded to establish that resveratrol binds to these extra-cellular receptors, these studies suggest that resveratrolcould activate several extracellular targets.

One of the very early in vitro events observed in tumorcells following resveratrol addition is an increase inintracellular calcium, which can be measured withinseconds after drug treatment (8). Calcium signaling(Fig. 1) may be a shared early response underlying thenontoxic feature of different anticancer natural products(17). If so, it may be possible to define a novel calcium-based mechanistic model for the development of a new

generation of nontoxic chemotherapeutics using resver-atrol as a model compound. However, the sentinel targetsinvolved in calcium activation are yet to be identified.Further, we have to recognize that some binding targetsfor resveratrol activate early events, whereas others mightpotentiate later events.

Despite important progress, the binding targets of resver-atrol that mediate its anticancer activity remain unknown.The relevant targets mediating effects in vitro versus in vivomight differ, and there seem to be several targets within asingle cell type as well as differences between cell types. Tobetter understand the mechanisms through which resver-atrol mediates its anticancer properties, studies need tobetter reflect conditions in vivo at drug levels that lead toinhibition of tumor growth.

© 2010 American Association for Cancer Research

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Fig. 1. Resveratrol-induced Ca2þ-mediated apoptosis. Resveratrol activates the inositol triphosphate receptor (IP3R) either through an extracellularreceptor or through an intracellular target resulting in release of extracellular receptor calcium, reflected in a rapid increase in intracellular calcium ([Ca2þ]i). Thesubsequent calcium uptake by the mitochondria causes mitochondrial membrane depolarization (Ym), increase in reactive oxygen species production(ROS), release of cytochrome c and smac/diablo, activating the intrinsic apoptotic pathway via caspases-9 and -3. Mitochondrial calcium-induced-calcium-release (mCICR) results in the activation of calpain, which then cleaves various substrates including calcium exchangers and pumps, likePMCA1, leading to additional elevation of intracellular calcium, further stimulating cell death.

Subramanian et al.

Clin Cancer Res; 16(24) December 15, 2010 Clinical Cancer Research5944




Resveratrol Actions

Is resveratrol antiproliferative, proapoptotic, andantiangiogenic?In vitro studies of resveratrol using a variety of tumor cell

lines have shown the drug to be antiproliferative as well asproapoptotic, through the activation of many differentpathways. Resveratrol inhibits cell-cycle progression bymodulating major cell-cycle regulators, arresting cells atthe G1/S, S, or G2/M phase, and by inhibiting DNA synth-esis. These effects, however, have not been shown in vivo.The mechanisms causing tumor inhibition, therefore, areunclear, even though tumor inhibition is observed in manydifferent animal models.Alternatively, it is well established that angiogenesis is

critical to tumor progression. Resveratrol has beenreported to have an antiangiogenic effect in certaintumor models, as indicated by decreased vessel densityin treated tumors (18–21). However, an observeddecrease in microvessel density cannot differentiatebetween cause and effect. Either the inhibition of tumorgrowth results in fewer blood vessels or an antiangio-genic effect causes decreased tumor outgrowth because ofa lack of nutrients and oxygen. The tumor-inhibitoryeffects in vivo could be due either to direct effects ofresveratrol on tumor cell growth or indirect effects onangiogenesis. An initial study in a mouse model of Lewislung carcinoma indicated a resveratrol effect on neovas-cularization (19). However, more in vivo studies areneeded to show a direct effect of resveratrol on bloodvessel formation as the mechanism for tumor inhibitionusing cell-specific reporter systems.In contrast to tumor inhibition, higher local doses of

resveratrol result in massive tumor cell death and tumorregression as indicated by terminal deoxynucleotidyl trans-ferasemediated dUTP nick end labeling (TUNEL) stainingand histology (11, 22). Thus, the reported proapoptoticeffects of resveratrol may only be relevant when localconcentrations of resveratrol more closely match the higherconcentrations studied in vitro. Therefore, strategies toimprove the bioavailability of resveratrol either systemi-cally or locally will provide more potent anticancer effects.

Bioavailability

How do we reconcile the anticancer effects ofresveratrol determined from in vitro studies within vivo measurements of resveratrol bioavailability?Preclinical studies using resveratrol in animal models

of different human cancers indicate that resveratrol haspotent antitumor properties. However, measurements ofserum levels of unmetabolized resveratrol suggest that itsbioavailability is very low after oral or intraperitonealadministration of the drug. This low bioavailability ismainly due to poor absorption and ready metabolism toglucuronidated and sulfated compounds, followed byrapid clearance of these metabolites. Serum levels ofunmetabolized resveratrol peak in the sub- to low-micro-

molar range (<3 mmol/L) within minutes of oral drugadministration, and decrease rapidly thereafter (11, 23–26). One implication of these findings is that the directactivation of cellular targets by resveratrol needs to reflectthis transient occurrence of both resveratrol and themetabolites. In addition, no accumulation of resveratrolin tumors or normal tissues has been found afterextended treatment regimens (11). These data contrastsharply with in vitro studies wherein sustained concentra-tions of tens of micromolars are required to obtain sub-stantive antitumor effects. This apparent conundrum wasdiscussed elegantly by Gescher and Steward (27). Theseobservations may suggest one of two different things: (a)the mechanisms of action of resveratrol are different invivo and in vitro; or (b) the in vitro conditions for growingcells somehow alter the sensitivity to resveratrol. In thefirst case, the antiangiogenic activity of resveratrol couldindirectly inhibit tumor growth. Such properties of thetumor environment are not recapitulated in vitro. In thesecond case, growth of tumor cells on a two-dimensionalsurface, the presence of high concentrations of growthfactors, high O2 levels, and lack of inhibitory cell-cellcontacts with surrounding normal cells could decreasethe sensitivity of cells to resveratrol. These optimizedgrowth conditions for tumor cells in culture may neces-sitate higher drug concentrations in vitro.

Reports of the relevant serum levels of resveratrol arefurther complicated by recent studies and unpublishedobservations indicating that resveratrol metabolites donot inhibit tumor cell growth (refs. 28, 29; and L. Sub-ramanian, unpublished observations). This finding sug-gests that glucuronidation and sulfation not only targetthe drug for removal from the body, but also mitigate itsantitumor activity. Minor conversion back to resveratrolby local sulfatases, for example, would not compensatefor this rapid decrease in activity. To increase the potencyof resveratrol treatment, it will be necessary to increasethe amount of unmetabolized compound at the tumorsite. A major obstacle to attaining this goal systemically isthe low solubility of the drug. Future research should bedirected toward new formulations and analogues thatincrease its bioavailability and delay its metabolism(Fig. 2).

The bioavailability conundrum also brings up the issueof resveratrol’s ability to differentiate between normalcells and cancerous cells in animal models. At low con-centrations in which there is no evidence of cell deatheither in normal or tumor tissue, there is, nevertheless, anantitumor effect of the drug leading to tumor inhibition.At higher local concentrations in which there is signifi-cant evidence for cell death in tumor tissue, the surround-ing normal tissue remains unaffected (11). Thisdiscrimination between normal and tumor cells couldarise due to differences in the cellular uptake of resver-atrol, the expression of cellular targets, or a differentialability to modify resveratrol into less active forms. Studiesof these processes are necessary to understand the non-toxic nature of resveratrol.






Consistencies between Disease Processes

Can the findings from resveratrol research done inthe realm of cancer be generalized tocardioprotection, life prolongation, diabetesprevention, and vice versa?

Resveratrol has a wide variety of pharmacologic activ-ities, making its study relevant to several diseases such as

cancer, cardiovascular and neurodegenerative diseases,and diabetes. However, it is not always clear how thesame drug causes different or even opposite effects indifferent cell types and in different diseases. For example,resveratrol causes cell death in tumor cells but is effec-tive as an antiapoptotic or protective agent in nervecells (30). Similarly, resveratrol is antiangiogenic incertain tumor models yet proangiogenic in the case of

© 2010 American Association for Cancer ResearchC R h

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Fig. 2. Translation of resveratrol to the clinic. Successful translation of resveratrol to an approved chemotherapy drug will necessitate new, moresoluble formulations to improve resveratrol bioavailability and delay metabolism to achieve significant improvements in antitumor efficacy.

Subramanian et al.





myocardial infarction (31, 32). Resveratrol increasesmitochondrial health in muscle cells, whereas it inducesmitochondria-mediated apoptosis in tumor cells (33).How might these effects occur? Although most directtargets of resveratrol may be expressed in a variety ofdifferent cell types, albeit at different levels, intracellularsignaling networks will be different depending on thecell type and cell status (nondividing, dividing, tumori-genic), leading to different outcomes following initialtarget interaction.Epidemiologic studies suggest a correlation between

diabetes and cancer risk. Resveratrol has been shown toexhibit antidiabetic effects. A recent report of the use of theantidiabetic drug metformin in cancer treatment suggeststhat resveratrol’s antidiabetic effects may be relevant to itsantitumor effects as well (32, 34). Therefore, findings ofresveratrol action will be of potential cross-disease impor-tance. As different molecular pathways are elucidated ingreater detail and are incorporated into larger networks, itwill become increasingly apparent how the addition of adrug like resveratrol can still be effective in very disparatediseases.Many of these diseases are prevalent simultaneously,

particularly in elderly patients. On the basis of currentliterature, it would seem that resveratrol could amelio-rate secondary conditions even as it is used to treat aprimary disease. However, it is necessary to carry outcomprehensive clinical trials on both the preventivepotential as well as curative efficacy of resvera-trol in different diseases before such claims can bevalidated.

Market Claims

What can we tell the public?Resveratrol has great potential as an anticancer drug

either as primary or as adjuvant therapy. One of its majorbenefits is a lack of toxicity at doses needed for itsantitumor effect. Toxicity remains one of the majorconcerns with current chemotherapies, and resveratrol,therefore, may allow for lowering of the doses oftoxic compounds while maintaining or enhancing theantitumor efficacy. Furthermore, there have been noreports of acquired resistance to resveratrol duringtreatment.At this time no clinical trials have been completed to

indicate that resveratrol will be effective as an anticancertreatment in humans. All the studies to date have beendone in tissue culture models or animal models. Thesestudies show a strong antitumor effect of resveratrol withoral dosing and the potential for even higher efficacy ifdoses can be further increased by overcoming deliveryand solubility obstacles. However, as has been shownwith numerous other potential drugs, results from ani-mal studies do not necessarily extrapolate to humans.Therefore, great caution has to be taken not to raise

expectations of success in using this compound as ananticancer agent before clinical trials have been success-fully conducted. Even as these trials proceed, the con-cerns raised in previous sections should be adequatelyaddressed in order to maximize resveratrol’s potential asa nontoxic anticancer drug. Currently, one trial is under-way in colon cancer patients, with trials for a number ofother types of cancer proposed. Ultimately, based on theresults of the clinical trials, the Food and Drug Admin-istration (FDA) alone has the authority to approve theuse of resveratrol in cancer therapy in the United States.In addition, its antioxidant effects, currently being testedin other clinical trials, may lead to its use as an approvedchemopreventive agent or in combination with otheranticancer drugs. Again, the nontoxic aspects of resver-atrol also make it a likely candidate for neoadjuvant oradjuvant treatment.

Nevertheless, resveratrol is already available as a nutri-tional supplement. Because nutraceuticals are moreloosely regulated by the FDA, much stronger claims aboutresveratrol’s benefits to human health and disease aretouted by this industry. Because those afflicted withcancer are desperate for a cure and are distrustful ofcurrent chemotherapy, many turn to nutraceutical pro-ducts, drawn by the promise of a successful "natural"treatment. Unfortunately, many of these claims aboutresveratrol go well beyond current research results, lead-ing to false hopes in patients. These expectations areincreased further by reports in newspapers and on tele-vision promoting resveratrol’s anticancer and anti-agingproperties along with a host of other health benefits. Allof these claims are yet to be scientifically proven inhumans. It is not surprising that companies with a com-mercial interest in resveratrol may oversell its qualities inorder to promote their product. However, it is importantthat researchers provide accurate information throughpeer-reviewed publications and to the public via newsoutlets and other avenues indicating the limitations of thestudies done to date. As results of human trials becomeknown, we will find out whether the initial promises canbe fulfilled.

Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were disclosed.

Acknowledgments

We thank Chue Vang for excellent technical assistance.

Grant Support

This work was supported by funds from the American Institute for CancerResearch (P.R. van Ginkel), the Mandelbaum Therapeutics Initiative (A.S.Polans), and the Retina Research Foundation (A.S. Polans is the M.D. MatthewsR.R.F. Professor).

Received 06/07/2010; accepted 09/13/2010; published OnlineFirst11/02/2010.






References1. Jang M, Cai L, Udeani GO, et al. Cancer chemopreventive activity of

resveratrol, a natural product derived from grapes. Science1997;275:218–20.

2. Maier-Salamon A, Hagenauer B, Wirth M, Gabor F, Szekeres T, JagerW. Increased transport of resveratrol across monolayers of the humanintestinal caco-2 cells is mediated by inhibition and saturation ofmetabolites. Pharm Res 2006;23:2107–15.

3. Kaldas MI, Walle UK, Walle T. Resveratrol transport and metabolismby human intestinal caco-2 cells. J Pharm Pharmacol 2003;55:307–12.

4. Lancon A, Delmas D, Osman H, Thenot JP, Jannin B, Latruffe N.Human hepatic cell uptake of resveratrol: Involvement of both passivediffusion and carrier-mediated process. Biochem Biophys Res Com-mun 2004;316:1132–7.

5. Murias M, Miksits M, Aust S, et al. Metabolism of resveratrol in breastcancer cell lines: Impact of sulfotransferase 1A1 expression on cellgrowth inhibition. Cancer Lett 2008;261:172–82.

6. Aggarwal BB, Bhardwaj A, Aggarwal RS, Seeram NP, Shishodia S,Takada Y. Role of resveratrol in prevention and therapy of cancer:Preclinical and clinical studies. Anticancer Res 2004;24:2783–840.

7. Pirola L, Frojdo S. Resveratrol: One molecule, many targets. IUBMBLife 2008;60:323–32.

8. Sareen D, Darjatmoko SR, Albert DM, Polans AS. Mitochondria,calcium, and calpain are key mediators of resveratrol-induced apop-tosis in breast cancer. Mol Pharmacol 2007;72:1466–75.

9. Feng L, Jin J, Zhang LF, Yan T, Tao WY. Analysis of the resveratrol-binding protein using phage-displayed random peptide library. ActaBiochim Biophys Sin (Shanghai) 2006;38:342–8.

10. Wang Z, Hsieh TC, Zhang Z, Ma Y, Wu JM. Identification andpurification of resveratrol targeting proteins using immobilized resver-atrol affinity chromatography. Biochem Biophys Res Commun2004;323:743–9.

11. van Ginkel PR, Sareen D, Subramanian L, et al. Resveratrol inhibitstumor growth of human neuroblastoma and mediates apoptosis bydirectly targeting mitochondria. Clin Cancer Res 2007;13:5162–9.

12. Beher D, Wu J, Cumine S, et al. Resveratrol is not a direct activator ofSIRT1 enzyme activity. Chem Biol Drug Des 2009;74:619–24.

13. Kaeberlein M, McDonagh T, Heltweg B, et al. Substrate-specificactivation of sirtuins by resveratrol. J Biol Chem 2005;280:17038–45.

14. Bass TM, Weinkove D, Houthoofd K, Gems D, Partridge L. Effects ofresveratrol on lifespan in drosophila melanogaster and caenorhabditiselegans. Mech Ageing Dev 2007;128:546–52.

15. Singh SK, Moretta D, Almaguel F, Wall NR, De Leon M, De Leon D.Differential effect of proIGF-II and IGF-II on resveratrol induced celldeath by regulating survivin cellular localization and mitochondrialdepolarization in breast cancer cells. Growth Factors 2007;25:363–72.

16. Lin HY, Lansing L, Merillon JM, et al. Integrin alphaVbeta3 contains areceptor site for resveratrol. FASEB J 2006;20:1742–4.

17. Das A, Banik NL, Ray SK. Flavonoids activated caspases for apop-tosis in human glioblastoma T98G and U87MG cells but not in humannormal astrocytes. Cancer 2010;116:164–76.

18. Garvin S, Ollinger K, Dabrosin C. Resveratrol induces apoptosis andinhibits angiogenesis in human breast cancer xenografts in vivo.Cancer Lett 2006;231:113–22.

19. Kimura Y, Okuda H. Resveratrol isolated from polygonum cuspidatumroot prevents tumor growth and metastasis to lung and tumor-induced neovascularization in lewis lung carcinoma-bearing mice. JNutr 2001;131:1844–9.

20. Chen JC, Chen Y, Lin JH, Wu JM, Tseng SH. Resveratrol suppressesangiogenesis in gliomas: Evaluation by color doppler ultrasound.Anticancer Res 2006;26:1237–45.

21. Tseng SH, Lin SM, Chen JC, et al. Resveratrol suppresses theangiogenesis and tumor growth of gliomas in rats. Clin Cancer Res2004;10:2190–202.

22. Zhou HB, Chen JJ, Wang WX, Cai JT, Du Q. Anticancer activity ofresveratrol on implanted human primary gastric carcinoma cells innude mice. World J Gastroenterol 2005;11:280–4.

23. Asensi M, Medina I, Ortega A, et al. Inhibition of cancer growth byresveratrol is related to its low bioavailability. Free Radic Biol Med2002;33:387–98.

24. Marier JF, Vachon P, Gritsas A, Zhang J, Moreau JP, Ducharme MP.Metabolism and disposition of resveratrol in rats: Extent of absorption,glucuronidation, and enterohepatic recirculation evidenced by alinked-rat model. J Pharmacol Exp Ther 2002;302:369–73.

25. Meng X, Maliakal P, Lu H, LeeMJ, Yang CS. Urinary and plasma levelsof resveratrol and quercetin in humans, mice, and rats after ingestionof pure compounds and grape juice. J Agric FoodChem 2004;52:935–42.

26. Yu C, Shin YG, Chow A, et al. Human, rat, and mouse metabolism ofresveratrol. Pharm Res 2002;19:1907–14.

27. Gescher AJ, Steward WP. Relationship between mechanisms, bioa-vailibility, and preclinical chemopreventive efficacy of resveratrol: Aconundrum. Cancer Epidemiol Biomarkers Prev 2003;12:953–7.

28. Wang LX, Heredia A, Song H, et al. Resveratrol glucuronides as themetabolites of resveratrol in humans: Characterization, synthesis, andanti-HIV activity. J Pharm Sci 2004;93:2448–57.

29. Miksits M, Wlcek K, Svoboda M, et al. Antitumor activity of resveratroland its sulfated metabolites against human breast cancer cells. PlantaMed 2009;75:1227–30.

30. Dasgupta B,Milbrandt J. Resveratrol stimulates AMP kinase activity inneurons. Proc Natl Acad Sci U S A 2007;104:7217–22.

31. Chen Y, Tseng SH. Review. Pro- and anti-angiogenesis effects ofresveratrol. In Vivo 2007;21:365–70.

32. Penumathsa SV, Maulik N. Resveratrol: A promising agent in promot-ing cardioprotection against coronary heart disease. Can J PhysiolPharmacol 2009;87:275–86.

33. Lagouge M, Argmann C, Gerhart-Hines Z, et al. Resveratrol improvesmitochondrial function and protects against metabolic disease byactivating SIRT1 and PGC-1alpha. Cell 2006;127:1109–22.

34. Hirsch HA, Iliopoulos D, Tsichlis PN, Struhl K. Metformin selectivelytargets cancer stem cells, and acts together with chemotherapy toblock tumor growth and prolong remission. Cancer Res 2009;69:7507–11.

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2010;16:5942-5948. Published OnlineFirst November 2, 2010.Clin Cancer Res Lalita Subramanian, Sherry Youssef, Saswati Bhattacharya, et al. Discussion

A Critical−−Resveratrol: Challenges in Translation to the Clinic

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