This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Holmes-McNary, M. Articles by Baldwin, A. S. Search for Related Content PubMed PubMed Citation Articles by Holmes-McNary, M. Articles by Baldwin, A. S., Jr.

Chemopreventive Properties of trans-Resveratrol Are Associated with Inhibition of Activation of the IB Kinase¹

Lineberger Comprehensive Cancer Center [M. H-M., A. S. B.] and Curriculum in Genetics and Molecular Biology [A. S. B.], University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7295

	ABSTRACT

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
trans-Resveratrol (Res), a phytoalexin found at high levels in grapes and in grape products such as red wine, has been shown to have anti-inflammatory and antioncogenic properties. Because the transcription factor nuclear factor B (NF-B) is involved in inflammatory diseases and oncogenesis, we tested whether Res could modulate NF-B activity. Res was shown to be a potent inhibitor of both NF-B activation and NF-B-dependent gene expression through its ability to inhibit IB kinase activity, the key regulator in NF-B activation, likely by inhibiting an upstream signaling component. In addition, Res blocked the expression of mRNA-encoding monocyte chemoattractant protein-1, a NF-B-regulated gene. Relative to cancer chemopreventive properties, Res induced apoptosis in fibroblasts after the induced expression of oncogenic H-Ras. Thus, Res is likely to function by inhibiting inflammatory and oncogenic diseases, at least in part, through the inhibition of NF-B activation by blocking IB kinase activity. These data may also explain aspects of the so-called "French paradox" that is associated with reduced mortality from coronary heart disease and certain cancers and provide a molecular rationale for the role of a potent chemopreventive compound in blocking the initiation of inflammation and oncogenesis.

	INTRODUCTION

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
Significant interest surrounds dietary approaches directed toward the prevention of disease initiation and progression. Res³ (trans-3, 4',5-trihydroxystilbene), a natural phytoalexin found in grapes and grape products such as red wine, has anticancer and anti-inflammatory effects (1 , 2) . These findings are consistent with epidemiological studies that defined the so-called "French paradox" (3 , 4) as the association of reduced mortality from coronary heart disease and breast cancer (4 , 5) with increased red wine consumption. In addition, Res was found to have both estrogenic/antiestrogenic activities in vitro and in vivo (6, 7, 8) and antioxidant properties (1 , 9, 10, 11, 12) . Recently, Res has been shown to possess chemopreventive activity by inhibiting cellular events associated with tumor initiation, promotion, and progression (1 , 13 , 14) ; by inhibiting ribonucleotide reductase (15) ; and by inhibiting proliferation of some cancer cells in vitro (8 , 13 , 16 , 17) . Pertinent to cancer prevention, Res also suppresses the expression of inducible nitric oxide synthase (17) and cyclooxygenase-2 (1 , 18) , which is likely to contribute to both its anti-inflammatory and antioncogenic mechanism. Despite these important advances, the molecular mechanism(s) by which Res exerts its broad biological effects has not yet been elucidated.

The transcription factor NF-B is strongly linked to inflammatory and immune responses (19, 20, 21, 22) and is associated with oncogenesis in certain models of cancer (23, 24, 25, 26, 27) . NF-B is important for the regulation of cell proliferation, cell transformation, and tumor development (28, 29, 30, 31) . Recently, we demonstrated that oncogenic forms of Ras (32) and the oncoprotein Bcr-Abl (26) both activate NF-B through the activation of the transcriptional function of the RelA/p65 subunit. Furthermore, not only is NF-B activity required for Ras to initiate cellular transformation, but it is also required for Bcr-Abl-initiated tumorigenesis and transformation (26 , 32) . Activated NF-B has been found in primary breast tumors (25 , 33) and has been shown to be required for proliferation and survival of Hodgkin’s disease tumor cells (24) . In terms of cell proliferation, breast cancer and other cancers often exhibit high levels of cyclin D1 (34 , 35) , and we and others have shown that NF-B activates transcription of the cyclin D1 gene (36 , 37) . Importantly, the requirement for NF-B in oncogenesis appears to be based, at least in part, on its ability to suppress transformation-associated apoptosis (23) .

NF-B activity is regulated in part by its subcellular localization. Under noninduced conditions, NF-B is sequestered in the cytoplasm through interactions with an inhibitor protein known as IB (28, 29, 30, 31) . Numerous extracellular stimuli can activate NF-B through signal transduction pathways that activate an IKK complex that phosphorylates IB on serines 32 and 36. The phosphorylation of IB leads to its ubiquitination and ultimate degradation by the proteasome (28, 29, 30, 31) , allowing NF-B to translocate to the nucleus where it activates the expression of genes. Activation of the NF-B/Rel family of transcription factors regulates the expression of genes that participate in pathways involving inflammation, cell proliferation, and apoptosis (28, 29, 30, 31) , including the inflammatory mediators nitric oxide synthase and cyclooxygenase-2 (38 , 39) . Although numerous effects have been described for Res, the molecular mechanisms responsible for its anti-inflammatory and antioncogenic effects are not yet clear. Here we asked whether the chemopreventive effect of Res occurs through inhibition of NF-B activation, and, if so, through what mechanism.

	MATERIALS AND METHODS

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
Cell Cultures and Cell Extracts
Human monocyte (THP-1) and macrophage (U937) cell lines were grown in RPMI 1640 supplemented with 10% fetal bovine serum, 5 x 10^-5 M ß-mercaptoethanol, and 1% penstrep. Cells were seeded at a density of 10⁶ cells/ml and cultured at 37°C in a 5% CO₂ atmosphere. Purified Res was purchased from Sigma (St. Louis, MO) and prepared according to the manufacturer’s protocol. Briefly, for all experiments, a 50 mg/ml stock solution prepared in 100% ethanol was used. Cells were preincubated for 60 min with Res (30 µM) and stimulated with either TNF (Promega; 10 ng/ml) for 15 min or with LPS (Promega; 1 µg/ml) for 4 h. CEs and NEs were prepared as described previously (40) and stored at -70°C. Rat1:iRas cultures expressing the IPTG-inducible oncogenic H-Ras allele (23) were cultured in DMEM supplemented with 10% fetal bovine serum. Before the experiments, subconfluent Rat1:iRas cell cultures were serum-starved for 4 h and either left untreated or stimulated with 5 mM IPTG in the presence or absence of Res.

EMSA
NEs (5–10 µg) were preincubated with 1 µg of poly(deoxyinosinic-deoxycytidylic acid) in binding buffer (10 mM Tris, 50 mM NaCl, 20% glycerol, 1 mM DTT, and 0.5 mM EDTA) for 10 min at room temperature. Approximately 30,000 cpm of ³²P-labeled DNA probe containing the murine MHC class I NF-B DNA binding site (41) were added and allowed to bind for 15 min. The complexes were separated by 5% PAGE and detected by autoradiography. Specificity of binding was examined by competition with excess unlabeled oligonucleotide (UV21). For supershift assays, NEs were incubated with antibodies against p50 and p65 subunits of NF-B for 20 min at room temperature before analysis by EMSA.

Transfection and Gene Expression Assay
3XB-Luc Assay.
THP-1 cells were cotransfected using DEAE/dextran with the empty expression vector (pDCR) and either the 3XB-luc reporter or the super-repressor form of IB (SR-IB) expression vector (42) . Transfections used 5.0 µg of the pDCR empty expression vector only, 1.0 µg of the 3XB-luc reporter, or 1.0 µg of the SR-IB expression vector and were brought to a final concentration of 5.0 µg with the empty vector. After 48 h, cells were pretreated for 60 min with Res (30 µM) and stimulated for 6 h with TNF (10 ng/ml). Cell lysates were made by freeze-thawing three times. Protein concentrations were determined, and 100 mg of protein were assayed for luciferase activity as described previously (32 , 43) . The results (Fig. 2A) are expressed as the fold luciferase induction relative to the transfection that contained the empty expression vector, whose value was placed at 1.0.

View larger version (23K): [in this window] [in a new window] [Download PPT slide]   Fig. 2. Res suppresses NF-B-regulated gene expression. A, THP-1 cells were cotransfected using DEAE/dextran with the empty expression vector (pDCR) and either the 3XB-luc reporter or the super-repressor form of IB (SR-IB) expression vector. To inhibit NF-B activity, SR-IB, which cannot be phosphorylated or degraded, was used to block nuclear translocation and subsequent transactivation of NF-B-responsive genes (42) . Transfections used 5.0 µg of the pDCR empty expression vector only, 1.0 µg of the 3XB-luc reporter, or 1.0 µg of the SR-IB expression vector and were brought to a final concentration of 5 µg with the empty vector. After 48 h, cells were prepared as described. The results are expressed as the fold luciferase induction relative to the transfection that contained the empty expression vector, whose value was placed at 1.0. Bars, mean ± SE determined from at least three independent transfection experiments. Statistical analysis was performed by ANOVA (StatView), and different letters between groups indicate significant difference at b, P < 0.01 and c, P < 0.05. B, THP-1 cells were cotransfected along with the pDCR empty expression vector as described above, except that either 1.0 µg of a NF-B CAT-linked reporter containing WT HIV-LTR-CAT or 1.0 µg of a MUT HIV-LTR-CAT in which both inducible B sites have been mutated. Pretreatment and stimulation were as described, cells were harvested, and CAT activity was determined. The results are expressed as the fold CAT induction as described above. Bars, mean ± SE determined from at least three independent transfection experiments. Statistical analysis was performed by ANOVA (StatView), and different letters between groups indicate significant difference at P > 0.01. C, THP-1 cells were pretreated with Res (30 µM), followed by TNF (10 ng/ml) stimulation for 3 h. Total RNA was reverse transcribed and amplified by PCR (RT-PCR) using specific primers for MCP-1 and actin. A representative photograph was scanned, and MCP-1 mRNA levels were quantitated by PhosphorImager analysis (Molecular Dynamics). Data are representative of three independent experiments.

	RT-PCR

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
THP-1 cells were pretreated with Res (30 µM), followed by TNF (10 ng/ml) stimulation for 3 h. RNA was isolated using the Trizol method (Life Technologies, Inc.), and 1 µg of total RNA was reverse-transcribed and amplified by PCR (RT-PCR) using specific primers for MCP-1 and actin (44) , as described previously. The oligonucleotide primers used were as follows: (a) MCP-1, 5'-GGCTGAGCCCACTTATCACTCATGG-3' (5' primer) and 5'-GGAAGCTTGCTGGAGGCGAGAGTGCGAG-3' (3' primer); and (b) actin, 5'-CCAACCGCGAGAAGATGACC-3' (5' primer) and 5'-GATCTTCATGAGGTAGTCAGT-3' (3' primer). Actin was used to determine equal protein loading. The PCR temperatures used were 94°C for 45 s, 55°C (actin) and 60°C (MCP-1) for 30 s, and 72°C for 90 s, followed by extension for 10 min at 72°C. The PCR reaction was set for various cycles (20–35 cycles) to maintain the linearity of the amplification. The PCR products (10 µl) were electrophoresed on a 2% agarose gel containing gel star fluorescent dye (FMC Corp., Philadelphia, PA). A representative photograph was scanned and analyzed. Negative controls consisted of tubes with and without RNA. MCP-1 mRNA levels were quantitated by PhosphorImager analysis (Molecular Dynamics). Data are representative of three independent experiments.

	Western Blotting

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
Equal amounts of CEs were resolved on a 10% SDS-polyacrylamide gel and transferred to nitrocellulose membrane. Blots were blocked in 5% milk in 1x TBST (Tris-buffered saline and 0.5% Tween 20) and probed with a specific IB antibody (1:1000; Rockland). Blots were probed with a secondary antirabbit antibody conjugated with horseradish peroxidase (1:10,000; Promega). Protein bands were visualized with an enhanced chemiluminescence detection system (ECL; Amersham Life Technologies).

	In Vitro Kinase Assay

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
THP-1 cells were treated as described above for the indicated time periods. Whole cell extracts were prepared, and IKK was immunoprecipitated with a specific antibody to the IKK-ß subunit. IKK activity was measured using a GST-IB-1–54(1–54) WT or a MUT GST-IB- substrate (45, 46, 47, 48, 49) . IKK activity was measured using a GST-IB-1–54(1–54) (4 µg) WT or a MUT GST-IB- substrate, where Ser³² and Ser³⁶ were substituted by Thr (4 µg; S32T and S36T; Refs. 45, 46, 47, 48, 49 ). These substrates were enzymatically phosphorylated by activated IKK with [-³²P]ATP (New England Nuclear, Boston, MA). IKK activity was quantitated by PhosphorImager analysis (Molecular Dynamics) and normalized to the IKK activity of untreated cells. Data are expressed as the fold induction. Data are representative of three independent experiments.

	ELISA in Situ Cell Death Assay and Microscopy

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
Rat1:iRas cells were pretreated with or without Res (30 µM) for 60 min before the addition of IPTG (5 mM) and incubated for 48 h. After incubation, cell death was detected by the ELISA in situ apoptosis assay (Boehringer Mannheim), and the percentage of apoptosis was quantitated. Staurosporine was used as the positive control for the induction of apoptosis. This assay measures DNA strand breaks and is therefore diagnostic for cells undergoing apoptosis. In a parallel experiment, Rat1:iRas cells were either pretreated with or without Res (30 µM) for 60 min before the addition of IPTG (5 mM) and incubated for 48 h. Nonadherent, dying cells are shown as refractive by phase-contrast microscopy.

	RESULTS

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 
Res Inhibits NF-B DNA Binding Activity.
For our initial studies, we investigated whether Res inhibited NF-B DNA binding activity. We used two macrophage/monocytic cell lines, THP-1 and U937, both of which are well characterized with regard to activation of NF-B. THP-1 cells were either left untreated or exposed to TNF, a potent inducer of NF-B in many cell types. As expected, TNF strongly induced activation of NF-B DNA binding activity, as determined by EMSA (Fig. 1 , Lane 2). In a dose-response study, we found that the effective dose of Res for inhibiting activation of NF-B was 30 µM (data not shown), and this dose was used for all experiments. Pretreatment with Res had little effect on the basal binding activity of NF-B (data not shown) but strongly blocked the ability of TNF to activate NF-B DNA binding (Fig. 1 , Lane 3). Similarly, the induction of NF-B in THP-1 cells by LPS, a potent bacterial endotoxin, was also effectively blocked by Res (Fig. 1 , Lane 5). Virtually identical data were obtained with U937 cells (data not shown), indicating that the ability of Res to block NF-B activation is not cell-type specific. Consistent with these results, it has been shown recently that Res can inhibit NF-B activation in RAW 264.7 macrophage cells (17) . In the same extracts, DNA binding of the constitutive transcription factor Oct-1 and the CAAT/enhancer binding protein transcription factor were unaffected by the presence of Res (data not shown), demonstrating that Res does not negatively affect transcription factors in a general manner.

View larger version (69K): [in this window] [in a new window] [Download PPT slide]   Fig. 1. Res-mediated repression of TNF- and LPS-induced NF-B activation. THP-1 cells were pretreated with Res (30 µM) and stimulated with either TNF (10 ng/ml; 15 min) or LPS (1 µg/ml; 4 h). NEs were analyzed by EMSA as described. Data are a photograph of an autoradiograph and are representative of three independent experiments.

Res Inhibits NF-B-dependent Gene Transcription.

Inhibition of TNF-induced IKK Activity.
The majority of inducers of NF-B stimulate a signal transduction pathway that leads to the activation of the IKK complex (45, 46, 47, 48, 49) that phosphorylates IB on Ser³² and Ser³⁶. Phosphorylated IB is then targeted for ubiquitination and subsequent degradation by the 26S proteasome, liberating NF-B and allowing nuclear translocation (28, 29, 30, 31) . To determine the level at which Res blocks NF-B activity, we analyzed the relative levels of IB after exposure of cells to TNF. CEs were prepared from the THP-1 cells used previously for EMSA. As expected, TNF stimulation led to a strong IB degradative response in THP-1 cells after 15 min (Fig. 3A , Lane 3), followed by a reappearance of IB at 30 min (Fig. 3A , Lane 4). Consistent with the ability to block NF-B activation (Fig. 1) , Res inhibited IB degradation in response to TNF (Fig. 3A , Lanes 6–8). Additionally, Res blocked TNF-induced phosphorylation of IB (data not shown) as demonstrated with the use of a phosphospecific antibody directed to Ser³², suggesting that Res inhibits an upstream signaling component in the TNF signaling pathway. We therefore analyzed levels of IKK activity after TNF treatment of THP-1 cells in the absence or presence of Res. Whole cell extracts were prepared, and IKK was immunoprecipitated with a specific antibody to the IKK-ß subunit (46) . IKK activity was measured using a GST-IB1–54(1–54) WT or a MUT GST-IB substrate, in which Ser³² and Ser³⁶ were substituted by threonine (S32T and S36T; Refs. 45, 46, 47, 48, 49 ). Res was found to be a potent inhibitor of inducible IKK activity in response to TNF exposure (Fig. 3B) . Additional data show that Res is apparently not an intrinsic IKK inhibitor because Res does not block IKK activity when added directly to the in vitro kinase reaction (data not shown). Moreover, it is unlikely that the mechanism of action of Res relative to its ability to inhibit NF-B acts through its antioxidant property because others (54) have shown that TNF-induced IKK activity was not affected by pretreatment with the potent antioxidant N-acetyl-L-cysteine. Additionally, it is unlikely that the mechanism of action of Res to block NF-B acts through its estrogenic properties because treatment of THP-1 cells with 17-ß-estradiol at concentrations as high as 10^-5 M did not lead to inhibition of NF-B (data not shown). Thus, our results suggest that the major mechanism whereby Res blocks NF-B activity acts through the inhibition of induction of IKK activity.

View larger version (39K): [in this window] [in a new window] [Download PPT slide]   Fig. 3. Res prevents TNF-induced degradation of IB by blocking IKK activity. A, IB degradation kinetics. THP-1 cells were either untreated (Lane 1), treated with TNF (10 ng/ml) for indicated times (Lanes 2–4), or preincubated for 60 min with Res (30 µM) followed by TNF (10 ng/ml) stimulation for the indicated times (Lanes 5–8). CEs were prepared, and protein expression was assessed by immunoblotting (60 µg/lane) with an antibody specific for IB (Rockland). THP-1 cells were treated as described. Whole cell extracts were immunoprecipitated with a specific antibody to the IKK-ß subunit (44) . IKK activity was measured using a GST-IB-(1–54) WT or a MUT GST-IB- substrate. IKK activity was measured using a GST-IB-(1–54) (4 µg) WT or a MUT GST-IB- substrate, in which Ser32 and Ser36 were substituted by Thr (4 µg; S32T and S36T). These substrates were enzymatically phosphorylated by activated IKK with [-32P]ATP. IKK activity was quantitated by PhosphorImager analysis (Molecular Dynamics) and normalized to the IKK activity of untreated cells. Data are expressed as the fold induction. Data are representative of three independent experiments.

Enhanced Apoptosis by Inhibition of Ras-mediated NF-B Activation.

View larger version (45K): [in this window] [in a new window] [Download PPT slide]   Fig. 4. Apoptotic effects of Res in Rat-1 cells expressing IPTG-inducible oncogenic H-Ras. A, Res -mediated suppression of NF-B in Rat1:iRas cells (23) . Rat1:iRas cultures stably expressing the IPTG-inducible oncogenic H-Ras allele were serum-starved, and after 4 h, complete media were added. Cells were pretreated in the absence (U, Lane 1) or presence of Res (30 µM; Lanes 3 and 4) for 60 min. Cultured Rat1:iRas cells were stimulated for 48 h with IPTG (5 mM; Lanes 2 and 3) to induce H-Ras-activated NF-B DNA binding. NEs were analyzed by EMSA with the MHC class I NF-B DNA binding probe as described previously (41) . Data are representative of three independent experiments. B, Res induces DNA fragmentation in Rat1:iRas cells. Rat1:iRas cells were pretreated with Res (30 µg/ml) for 60 min before the addition of IPTG (5 mM) and incubated for 48 h. After incubation, cell death was detected by the ELISA in situ apoptosis assay (Boehringer Mannheim), and the percentage of apoptosis was quantitated. Staurosporine was used as a positive control for the induction of apoptosis. C, Res enhances apoptosis in IPTG-induced Rat1:iRas cells. Micrograph panel (x20 magnification): Rat1:iRas cells were either pretreated without (U) or with Res (Res; 30 µM) for 60 min before the addition of IPTG (5 mM; iptg and I+Res) and incubated for 48 h. Nonadherent dying cells are shown as refractive by phase-contrast microscopy.

	DISCUSSION

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

Our data indicate that Res is a potent inhibitor of NF-B nuclear translocation and IB degradation. Furthermore, Res effects are mediated through the inhibition of IKK, the key regulatory complex required for NF-B activation of gene transcription. The molecular target of Res action is presently unknown because Res does not appear to directly block IKK activity. Presumably, Res inhibits an upstream signaling component that leads to the activation of IKK, and we are presently examining whether Res inhibits NF-B-inducing kinase or MEKK1, upstream activators of the IKK complex (46 , 64, 65, 66, 67) . Because evidence has been presented that Res can block AP-1 activity (18) , which is consistent with our observations (data not shown), one possibility is that MEKK1 is the target of Res action because MEKK1 can activate both the AP-1 and NF-B pathways (46 , 64, 65, 66, 67) . This hypothesis, if proven, could explain the dual inhibition of NF-B and of AP-1 transcriptional responses.

NF-B is strongly associated with inflammatory diseases and oncogenesis (28, 29, 30, 31) , and the activation of NF-B target genes, including proinflammatory cytokines, has been implicated in promoting the transformation and survival of tumor cells (23 , 42 , 68, 69, 70) . However, NF-B activation has been shown to be blocked by anti-inflammatory compounds such as aspirin (71 , 72) and glucocorticoids (73, 74, 75, 76) . Therefore, our results demonstrate that a potent chemopreventive compound also targets NF-B activation to block both inflammation and cancer initiation. In this case, Res blocks the signaling pathway leading to NF-B activation through its ability to block IKK activation. Res would then block the expression of NF-B-dependent genes such as MCP-1 and other genes that would normally promote inflammation, protect against apoptosis, and potentiate cell growth. Thus, our results provide a molecular rationale to explain the broad chemopreventive properties of Res.

	ACKNOWLEDGMENTS

 
We thank the members of the Baldwin laboratory for helpful discussion; Drs. M. Mayo, S. Westerheide, J. Webster-Cyriaque, and F. L. McNary for careful reading of the manuscript; Drs. J. Norris, M. Mayo, B. Ashburner, and C. Jobin for providing the NIH-3T3 and Rat1:iRas cells, SR-IB-, Gal-4-p65, and HIV-LTR-CAT plasmids, and PCR primers; Dr. Frank Mercurio (Signal Pharmaceuticals, Inc., San Diego, CA) for generously providing the GST- WT and MUT plasmids and IKK-specific antibody; and Dr. R. Bagnell and V. Madden for microscopy services and assistance.

	FOOTNOTES

 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Supported in part by National Cancer Institute Grants CA72771 and CA75080 (to A. S. B.), North Carolina Affiliate-American Heart Association Fellowship 9894835U (to M. H-M.), and National Cancer Institute-National Research Service Award 1F32CA77908-01 (to M. H-M).

² To whom requests for reprints should be addressed, at Lineberger Comprehensive Cancer Center, CB# 7295, University of North Carolina, Chapel Hill, NC 27599-7295. Phone: (919) 966-3652; Fax: (919) 966–0444; E-mail: jhall{at}med.unc.edu

³ The abbreviations used are: Res, trans-resveratrol; NF-B, nuclear factor B; IKK, IB kinase; MCP-1, monocyte chemoattractant protein-1; TNF, tumor necrosis factor; LPS, lipopolysaccharide; LTR, long terminal repeat; CAT, chloramphenicol aminotransferase; GST, glutathione S-transferase; WT, wild-type; MUT, mutant; IPTG, isopropyl-ß-D-thiogalactopyranoside; MEKK1, mitogen-activated protein/extracellular signal-regulated kinase kinase kinase 1; AP-1, activator protein 1; CE, cytoplasmic extract; NE, nuclear extract; SR-IB, NF-B IB inhibitor; IB, inhibitor B; EMSA, electrophoretic mobility shift assay; RT-PCR, reverse transcription-PCR.

Received 11/17/99. Accepted 5/ 2/00.

	REFERENCES

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RT-PCR Western Blotting In Vitro Kinase Assay ELISA in Situ Cell... RESULTS DISCUSSION REFERENCES

 

1. Jang M., Cai L., Udeani G. O., Slowing K. V., Thomas C. F., Beecher C. W., Fong H. H., Farnsworth N. R., Kinghorn A. D., Mehta R. G., Moon R. C., Pezzuto J. M. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science (Washington DC), 275: 218-220, 1997.[Abstract/Free Full Text]
2. Soleas G. J., Diamandis E. P., Goldberg D. M. Wine as a biological fluid: history, production, and role in disease prevention. J. Clin. Lab. Anal., 11: 287-313, 1997.[Medline]
3. Richard J. L. Coronary risk factors. The French paradox. Arch. Mal Coeur Vaiss., 80: 17-21, 1987.
4. Renaud S., de Lorgeril M. The French paradox: dietary factors and cigarette smoking-related health risks. Ann. N. Y. Acad. Sci., 686: 299-309, 1993.[Medline]
5. Gronbaek M., Deis A., Sorensen T. I., Becker U., Schnohr P., Jensen G. Mortality associated with moderate intakes of wine, beer, or spirits. Br. Med. J., 310: 1165-1169, 1995.[Abstract/Free Full Text]
6. Gehm B. D., McAndrews J. M., Chien P. Y., Jameson J. L. Resveratrol, a polyphenolic compound found in grapes and wine, is an agonist for the estrogen receptor. Proc. Natl. Acad. Sci. USA, 94: 14138-14143, 1997.[Abstract/Free Full Text]
7. Lu R., Serrero G. Resveratrol, a natural product derived from grape, exhibits antiestrogenic activity and inhibits the growth of human breast cancer cells. J. Cell. Physiol., 179: 297-304, 1999.[Medline]
8. Basly J-P., Marre-Fournier F., Le Bail J-C., Habrioux G., Chulia A. J. Estrogenic/antiestrogenic and scavenging properties of (E)- and (Z)-resveratrol. Life Sci., 66: 769-777, 2000.[Medline]
9. Goldberg D. M. More on antioxidant activity of resveratrol in red wine. Clin. Chem., 42: 113-114, 1996.[Free Full Text]
10. Kawada N., Seki S., Inoue M., Kuroki T. Effect of antioxidants, resveratrol, quercetin, and N-acetylcysteine on the functions of cultured rat hepatic stellate cells and Kupffer cells. Hepatology, 27: 1265-1274, 1998.[Medline]
11. Waffo Teguo P., Fauconneau B., Deffieux G., Huguet F., Vercauteren J., Merillon J. M. Isolation, identification, and antioxidant activity of three stilbene glucosides newly extracted from Vitis vinifera cell cultures. J. Nat. Prod., 61: 655-657, 1998.[Medline]
12. Ray P. S., Maulik G., Cordis G. A., Bertelli A. A., Bertelli A., Das D. K. The red wine antioxidant resveratrol protects isolated rat hearts from ischemia reperfusion injury. Free Radical Biol. Med., 27: 160-169, 1999.[Medline]
13. Mgbonyebi O. P., Russo J., Russo I. H. Antiproliferative effect of synthetic resveratrol on human breast epithelial cells. Int. J. Oncol., 12: 865-869, 1998.[Medline]
14. Carbo N., Costelli P., Baccino F. M., Lopez-Soriano F. J., Argiles J. M. Resveratrol, a natural product present in wine, decreases tumour growth in a rat tumour model. Biochem. Biophys. Res. Commun., 254: 739-743, 1999.[Medline]
15. Fontecave M., Lepoivre M., Elleingand E., Gerez C., Guittet O. Resveratrol, a remarkable inhibitor of ribonucleotide reductase. FEBS Lett., 421: 277-279, 1998.[Medline]
16. ElAttar T. M., Virji A. S. Modulating effect of resveratrol and quercetin on oral cancer cell growth and proliferation. Anticancer Drugs, 10: 187-193, 1999.[Medline]
17. Tsai S. H., Lin-Shiau S. Y., Lin J. K. Suppression of nitric oxide synthase and the down-regulation of the activation of NF-B in macrophages by resveratrol. Br. J. Pharmacol., 126: 673-680, 1999.[Medline]
18. Subbaramaiah K., Chung W. J., Michaluart P., Telang N., Tanabe T., Inoue H., Jang M., Pezzuto J. M., Dannenberg A. J. Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells. J. Biol. Chem., 273: 21875-21882, 1998.[Abstract/Free Full Text]
19. Barnes P. J., Karin M. Nuclear factor-B: a pivotal transcription factor in chronic inflammatory diseases. N. Engl. J. Med., 336: 1066-1071, 1997.[Free Full Text]
20. Brand K., Page S., Walli A. K., Neumeier D., Baeuerle P. A. Role of nuclear factor-B in atherogenesis. Exp. Physiol., 82: 297-304, 1997.[Abstract]
21. Jobin C., Panja A., Hellerbrand C., Iimuro Y., Didonato J., Brenner D. A., Sartor R. B. Inhibition of proinflammatory molecule production by adenovirus-mediated expression of a nuclear factor B super-repressor in human intestinal epithelial cells. J. Immunol., 160: 410-418, 1998.[Abstract/Free Full Text]
22. Miagkov A. V., Kovalenko D. V., Brown C. E., Didsbury J. R., Cogswell J. P., Stimpson S. A., Baldwin A. S., Makarov S. S. NF-B activation provides the potential link between inflammation and hyperplasia in the arthritic joint. Proc. Natl. Acad. Sci. USA, 95: 13859-13864, 1998.[Abstract/Free Full Text]
23. Mayo M. W., Wang C. Y., Cogswell P. C., Rogers-Graham K. S., Lowe S. W., Der C. J., Baldwin A. S., Jr. Requirement of NF-B activation to suppress p53-independent apoptosis induced by oncogenic Ras. Science (Washington DC), 278: 1812-1815, 1997.[Abstract/Free Full Text]
24. Bargou R. C., Emmerich F., Krappmann D., Bommert K., Mapara M. Y., Arnold W., Royer H. D., Grinstein E., Greiner A., Scheidereit C., Dorken B. Constitutive nuclear factor-B-RelA activation is required for proliferation and survival of Hodgkin’s disease tumor cells. J. Clin. Invest., 100: 2961-2969, 1997.[Medline]
25. Sovak M. A., Bellas R. E., Kim D. W., Zanieski G. J., Rogers A. E., Traish A. M., Sonenshein G. E. Aberrant nuclear factor-B/Rel expression and the pathogenesis of breast cancer. J. Clin. Invest., 100: 2952-2960, 1997.[Medline]
26. Reuther J. Y., Reuther G. W., Cortez D., Pendergast A. M., Baldwin A. S., Jr. A requirement for NF-B activation in Bcr-Abl-mediated transformation. Genes Dev., 12: 968-981, 1998.[Abstract/Free Full Text]
27. Krappmann D., Emmerich F., Kordes U., Scharschmidt E., Dorken B., Scheidereit C. Molecular mechanisms of constitutive NF-B/Rel activation in Hodgkin/Reed-Sternberg cells. Oncogene, 18: 943-953, 1999.[Medline]
28. Baldwin A. S., Jr. The NF-B and IB proteins: new discoveries and insights. Annu. Rev. Immunol., 14: 649-683, 1996.[Medline]
29. Verma I. M., Stevenson J. IB kinase: beginning, not the end. Proc. Natl. Acad. Sci. USA, 94: 11758-11760, 1997.[Free Full Text]
30. Maniatis T. Catalysis by a multiprotein IB kinase complex. Science (Washington DC), 278: 818-819, 1997.[Free Full Text]
31. Ghosh S., May M. J., Kopp E. B. NF-B and Rel proteins: evolutionarily conserved mediators of immune responses. Annu. Rev. Immunol., 16: 225-260, 1998.[Medline]
32. Finco T. S., Westwick J. K., Norris J. L., Beg A. A., Der C. J., Baldwin A. S., Jr. Oncogenic Ha-Ras-induced signaling activates NF-B transcriptional activity, which is required for cellular transformation. J. Biol. Chem., 272: 24113-24116, 1997.[Abstract/Free Full Text]
33. Nakshatri H., Bhat-Nakshatri P., Martin D. A., Goulet R. J., Jr., Sledge G. W., Jr. Constitutive activation of NF-B during progression of breast cancer to hormone-independent growth. Mol. Cell. Biol., 17: 3629-3639, 1997.[Abstract]
34. Sasano H., Frost A. R., Saitoh R., Taniyama Y., Nagura H., Matsunaga G., Takehana K., Kimura M., Silverberg S. G. Immunolocalization of cyclins D and E and cyclin-dependent kinase (cdk) 2 and 4 in human breast carcinoma. Anticancer Res., 17: 3685-3690, 1997.[Medline]
35. Hedberg Y., Davoodi E., Roos G., Ljungberg B., Landberg G. Cyclin-D1 expression in human renal-cell carcinoma. Int. J. Cancer, 84: 268-272, 1999.[Medline]
36. Guttridge D. C., Albanese C., Reuther J. Y., Pestell R. G., Baldwin A. S., Jr. NF-B controls cell growth and differentiation through transcriptional regulation of cyclin D1. Mol. Cell. Biol., 19: 5785-5799, 1999.[Abstract/Free Full Text]
37. Hinz M., Krappmann D., Eichten A., Heder A., Scheidereit C., Strauss M. NF-B function in growth control: regulation of cyclin D1 expression and G₀/G₁-to-S-phase transition. Mol. Cell. Biol., 19: 2690-2698, 1999.[Abstract/Free Full Text]
38. Xie Q. W., Kashiwabara Y., Nathan C. Role of transcription factor NF-B/Rel in induction of nitric oxide synthase. J. Biol. Chem., 269: 4705-4708, 1994.[Abstract/Free Full Text]
39. Yamamoto K., Arakawa T., Ueda N., Yamamoto S. Transcriptional roles of nuclear factor B and nuclear factor interleukin-6 in tumor necrosis factor -dependent induction of cyclooxygenase-2 in MC3T3–E1 cells. J. Biol. Chem., 270: 31315-31350, 1995.[Abstract/Free Full Text]
40. Finco T. S., Beg A. A., Baldwin A. S., Jr. Inducible phosphorylation of IB is not sufficient for its dissociation from NF-B and is inhibited by protease inhibitors. Proc. Natl. Acad. Sci. USA, 91: 11884-11888, 1994.[Abstract/Free Full Text]
41. Cogswell P. C., Mayo M. W., Baldwin A. S., Jr. Involvement of Egr-1/RelA synergy in distinguishing T cell activation from tumor necrosis factor--induced NF-B1 transcription. J. Exp. Med., 185: 491-497, 1997.[Abstract/Free Full Text]
42. Wang C. Y., Mayo M. W., Baldwin A. S., Jr. TNF- and cancer therapy-induced apoptosis: potentiation by inhibition of NF-B. Science (Washington DC), 274: 784-787, 1996.[Abstract/Free Full Text]
43. Norris J. L., Baldwin A. S., Jr. Oncogenic Ras enhances BF-kB transcriptional activity through Raf-dependent and Raf-independent mitogen-activated protein kinase signaling pathways. J. Biol. Chem., 274: 13841-13846, 1999.[Abstract/Free Full Text]
44. Boring L., Gosling J., Cleary M., Charo I. F. Decreased lesion formation in CCR2-/- mice reveals a role for chemokines in the initiation of atherosclerosis. Nature (Lond.), 394: 894-897, 1998.[Medline]
45. DiDonato J. A., Hayakawa M., Rothwarf D. M., Zandi E., Karin M. A cytokine-responsive IB kinase that activates the transcription factor NF-B. Nature (Lond.), 388: 548-554, 1997.[Medline]
46. Mercurio F., Zhu H., Murray B. W., Shevchenko A., Bennett B. L., Li J., Young D. B., Barbosa M., Mann M., Manning A., Rao A. IKK-1 and IKK-2: cytokine-activated IB kinases essential for NF-B activation. Science (Washington DC), 278: 860-866, 1997.[Abstract/Free Full Text]
47. Woronicz J. D., Gao X., Cao Z., Rothe M., Goeddel D. V. IB kinase-ß. NF-B activation and complex formation with IB kinase- and NIK. Science (Washington DC), 278: 866-869, 1997.[Abstract/Free Full Text]
48. Regnier C. H., Song H. Y., Goeddel D. V., Cao Z., Rothe M. Identification and characrterization of an IB kinase. Cell, 90: 373-383, 1997.[Medline]
49. Zandi E., Rothwarf D. M., Delhase M., Hayakawa M., Karin M. The IB kinase complex (IKK) contains two kinase subunits. IKK and IKKß, necessary for IB phosphorylation and NF-B activation. Cell, 91: 243-252, 1997.[Medline]
50. Brockman J. A., Scherer D. C., McKinsey T. A., Hall S. M., Qi X., Lee W. Y., Ballard D. W. Coupling of a signal response domain in IB to multiple pathways for NF-B activation. Mol. Cell. Biol., 15: 2809-2818, 1995.[Abstract]
51. Brown K., Gerstenberger S., Carlson L., Franzoso G., Siebenlist U. Control of IB proteolysis by site-specific, signal-induced phosphorylation. Science (Washington DC), 267: 1485-1491, 1995.[Abstract/Free Full Text]
52. Traenckner E. B., Pahl H. L., Henkel T., Schmidt K. N., Wilk S., Baeuerle P. A. Phosphorylation of human IB- on serines 32 and 36 controls IB- proteolysis and NF-B activation in response to diverse stimuli. EMBO J., 14: 2876-2883, 1995.[Medline]
53. Chen Z., Hagler J., Palombella V. J., Melandri F., Scherer D., Ballard D., Maniatis T. Signal-induced site-specific phosphorylation targets IB to the ubiquitin-proteasome pathway. Genes Dev., 9: 1586-1597, 1995.[Abstract/Free Full Text]
54. Li N., Karin M. Is NF-B the sensor of oxidative stress?. FASEB J., 13: 1137-1143, 1999.[Abstract/Free Full Text]
55. Finco T. S., Baldwin A. S. Mechanistic aspects of NF-B regulation: the emerging role of phosphorylation and proteolysis. Immunity, 3: 263-272, 1995.[Medline]
56. Block G., Patterson B., Subar A. Fruit, vegetables, and cancer prevention: a review of the epidemiological evidence. Nutr. Cancer, 18: 1-29, 1992.[Medline]
57. Willett W. C. Diet and health: what should we eat?. Science (Washington DC), 264: 532-537, 1994.[Abstract/Free Full Text]
58. Hu F. B., Stampfer M. J., Manson J. E., Rimm E., Colditz G. A., Rosner B. A., Hennekens C. H., Willett W. C. Dietary fat intake and the risk of coronary heart disease in women. N. Engl. J. Med., 337: 1491-1499, 1997.[Abstract/Free Full Text]
59. Li D., Wang M., Paul G. P., Pitot H. C., Dragan Y. Dietary oat lipids-induced novel DNA modifications and suppression of altered hepatic foci formation. Nutr. Cancer, 33: 40-45, 1999.[Medline]
60. Constant J. Alcohol, ischemic heart disease, and the French paradox. Coron. Artery Dis., 8: 645-649, 1997.[Medline]
61. Kopp P. Resveratrol, a phytoestrogen found in red wine. A possible explanation for the conundrum of the "French paradox"?. Eur. J. Endocrinol., 138: 619-620, 1998.[Medline]
62. Wattenberg L. W. Inhibition of carcinogenic effects of polycyclic hydrocarbons by benzyl isothiocyanate and related compounds. J. Natl. Cancer Inst., 58: 395-398, 1977.
63. Clinton S. K., Giovannucci E. Diet, nutrition, and prostate cancer. Annu. Rev. Nutr., 18: 413-440, 1998.[Medline]
64. Lee F. S., Hagler J., Chen Z. J., Maniatis T. Activation of the IB kinase complex by MEKK1, a kinase of the JNK pathway. Cell, 88: 213-222, 1997.[Medline]
65. Karin M., Delhase M. JNK or IKK, AP-1 or NF-B, which are the targets for MEK kinase 1 action?. Proc. Natl. Acad. Sci. USA, 95: 9067-9069, 1998.[Free Full Text]
66. Ling L., Cao Z., Goeddel D. V. NF-B-inducing kinase activates IKK- by phosphorylation of Ser-176. Proc. Natl. Acad. Sci. USA, 95: 3792-3797, 1998.[Abstract/Free Full Text]
67. Nakano H., Shindo M., Sakon S., Nishinaka S., Mihara M., Yagita H., Okumura K. Differential regulation of IB kinase and ß by two upstream kinases, NF-B-inducing kinase and mitogen-activated protein kinase/ERK kinase kinase-1. Proc. Natl. Acad. Sci. USA, 95: 3537-3542, 1998.[Abstract/Free Full Text]
68. Dong G., Chen Z., Kato T., Van Waes C. The host environment promotes the constitutive activation of nuclear factor-B and proinflammatory cytokine expression during metastatic tumor progression of murine squamous cell carcinoma. Cancer Res., 59: 3495-3504, 1999.[Abstract/Free Full Text]
69. Gilmore T. D., Koedood M., Piffat K. A., White D. W. Rel/NF-B/IB proteins and cancer. Oncogene, 13: 1367-1378, 1996.[Medline]
70. Van Antwerp D. J., Martin S. J., Kafri T., Green D. R., Verma I. M. Suppression of TNF--induced apoptosis by NF-B. Science (Washington DC), 274: 787-789, 1996.[Abstract/Free Full Text]
71. Kopp E., Ghosh S. Inhibition of NF-B by sodium salicylate and aspirin. Science (Washington DC), 265: 956-959, 1994.[Abstract/Free Full Text]
72. Yin M. J., Yamamoto Y., Gaynor R. B. The anti-inflammatory agents aspirin and salicylate inhibit the activity of IB kinase-ß. Nature (Lond.), 396: 77-80, 1998.[Medline]
73. Scheinman R. I., Gualberto A., Jewell C. M., Cidlowski J. A., Baldwin A. S., Jr. Characterization of mechanisms involved in transrepression of NF-B by activated glucocorticoid receptors. Mol. Cell. Biol., 15: 943-953, 1995.[Abstract]
74. Scheinman R. I., Cogswell P. C., Lofquist A. K., Baldwin A. S., Jr. Role of transcriptional activation of IB in mediation of immunosuppression by glucocorticoids. Science (Washington DC), 270: 283-286, 1995.[Abstract/Free Full Text]
75. Auphan N., DiDonato J. A., Rosette C., Helmberg A., Karin M. Immunosuppression by glucocorticoids: inhibition of NF-B activity through induction of IB synthesis. Science (Washington DC), 270: 286-290, 1995.[Abstract/Free Full Text]
76. Caldenhoven E., Liden J., Wissink S., Van de Stolpe A., Raaijmakers J., Koenderman L., Okret S., Gustafsson J. A., Van der Saag P. T. Negative cross-talk between RelA and the glucocorticoid receptor: a possible mechanism for the antiinflammatory action of glucocorticoids. Mol. Endocrinol., 9: 401-412, 1995.[Abstract/Free Full Text]

This article has been cited by other articles:

				M. N. Chahine and G. N. Pierce Therapeutic Targeting of Nuclear Protein Import in Pathological Cell Conditions Pharmacol. Rev., September 1, 2009; 61(3): 358 - 372. [Abstract] [Full Text] [PDF]

				D. Zhang, L. Qiu, X. Jin, Z. Guo, and C. Guo Nuclear Factor-{kappa}B Inhibition by Parthenolide Potentiates the Efficacy of Taxol in Non-Small Cell Lung Cancer In vitro and In vivo Mol. Cancer Res., July 1, 2009; 7(7): 1139 - 1149. [Abstract] [Full Text] [PDF]

				A. Sharafkhaneh, S. Velamuri, V. Badmaev, C. Lan, and N. Hanania Review: The potential role of natural agents in treatment of airway inflammation Therapeutic Advances in Respiratory Disease, December 1, 2007; 1(2): 105 - 120. [Abstract] [PDF]

				T. Szkudelski Resveratrol-induced inhibition of insulin secretion from rat pancreatic islets: evidence for pivotal role of metabolic disturbances Am J Physiol Endocrinol Metab, October 1, 2007; 293(4): E901 - E907. [Abstract] [Full Text] [PDF]

				E.-O. Lee, H.-J. Lee, H.-S. Hwang, K.-S. Ahn, C. Chae, K.-S. Kang, J. Lu, and S.-H. Kim Potent inhibition of Lewis lung cancer growth by heyneanol A from the roots of Vitis amurensis through apoptotic and anti-angiogenic activities Carcinogenesis, October 1, 2006; 27(10): 2059 - 2069. [Abstract] [Full Text] [PDF]

				J. K. Kundu, Y. K. Shin, S. H. Kim, and Y.-J. Surh Resveratrol inhibits phorbol ester-induced expression of COX-2 and activation of NF-{kappa}B in mouse skin by blocking I{kappa}B kinase activity Carcinogenesis, July 1, 2006; 27(7): 1465 - 1474. [Abstract] [Full Text] [PDF]

				J. Mohan, A. A. Gandhi, B. C. Bhavya, R. Rashmi, D. Karunagaran, R. Indu, and T. R. Santhoshkumar Caspase-2 Triggers Bax-Bak-dependent and -independent Cell Death in Colon Cancer Cells Treated with Resveratrol J. Biol. Chem., June 30, 2006; 281(26): 17599 - 17611. [Abstract] [Full Text] [PDF]

				M. Sengottuvelan, P. Viswanathan, and N. Nalini Chemopreventive effect of trans-resveratrol - a phytoalexin against colonic aberrant crypt foci and cell proliferation in 1,2-dimethylhydrazine induced colon carcinogenesis Carcinogenesis, May 1, 2006; 27(5): 1038 - 1046. [Abstract] [Full Text] [PDF]

				K. R. Martin Targeting Apoptosis with Dietary Bioactive Agents Experimental Biology and Medicine, February 1, 2006; 231(2): 117 - 129. [Abstract] [Full Text] [PDF]

				J. E. Hoberg, A. E. Popko, C. S. Ramsey, and M. W. Mayo I{kappa}B Kinase {alpha}-Mediated Derepression of SMRT Potentiates Acetylation of RelA/p65 by p300 Mol. Cell. Biol., January 15, 2006; 26(2): 457 - 471. [Abstract] [Full Text] [PDF]

				C. A. Conover, L. K. Bale, S. C. Harrington, Z. T. Resch, M. T. Overgaard, and C. Oxvig Cytokine stimulation of pregnancy-associated plasma protein A expression in human coronary artery smooth muscle cells: inhibition by resveratrol Am J Physiol Cell Physiol, January 1, 2006; 290(1): C183 - C188. [Abstract] [Full Text] [PDF]

				H. S. Youn, J. Y. Lee, K. A. Fitzgerald, H. A. Young, S. Akira, and D. H. Hwang Specific Inhibition of MyD88-Independent Signaling Pathways of TLR3 and TLR4 by Resveratrol: Molecular Targets Are TBK1 and RIP1 in TRIF Complex J. Immunol., September 1, 2005; 175(5): 3339 - 3346. [Abstract] [Full Text] [PDF]

				S. Ghisletti, C. Meda, A. Maggi, and E. Vegeto 17{beta}-Estradiol Inhibits Inflammatory Gene Expression by Controlling NF-{kappa}B Intracellular Localization Mol. Cell. Biol., April 15, 2005; 25(8): 2957 - 2968. [Abstract] [Full Text] [PDF]

				H. Jiang, L. Zhang, J. Kuo, K. Kuo, S. C. Gautam, L. Groc, A. I. Rodriguez, D. Koubi, T. Jackson Hunter, G. B. Corcoran, et al. Resveratrol-induced apoptotic death in human U251 glioma cells Mol. Cancer Ther., April 1, 2005; 4(4): 554 - 561. [Abstract] [Full Text] [PDF]

				M. T. Yip-Schneider, H. Nakshatri, C. J. Sweeney, M. S. Marshall, E. A. Wiebke, and C. M. Schmidt Parthenolide and sulindac cooperate to mediate growth suppression and inhibit the nuclear factor-{kappa}B pathway in pancreatic carcinoma cells Mol. Cancer Ther., April 1, 2005; 4(4): 587 - 594. [Abstract] [Full Text] [PDF]

				P. Marchetti Natural medicine: a 'new frontier' in oncology? Ann. Onc., March 1, 2005; 16(3): 339 - 340. [Full Text] [PDF]

				M. Lun, P. L. Zhang, P. K. Pellitteri, A. Law, T. L. Kennedy, and R. E. Brown Nuclear Factor-kappaB Pathway as a Therapeutic Target in Head and Neck Squamous Cell Carcinoma: Pharmaceutical and Molecular Validation in Human Cell Lines Using Velcade and siRNA/NF-{kappa}B. Ann. Clin. Lab. Sci., January 1, 2005; 35(3): 251 - 258. [Abstract] [Full Text] [PDF]

				J.-L. Su, M.-T. Lin, C.-C. Hong, C.-C. Chang, S.-G. Shiah, C.-W. Wu, S.-T. Chen, Y.-P. Chau, and M.-L. Kuo Resveratrol induces FasL-related apoptosis through Cdc42 activation of ASK1/JNK-dependent signaling pathway in human leukemia HL-60 cells Carcinogenesis, January 1, 2005; 26(1): 1 - 10. [Abstract] [Full Text] [PDF]

				L. E. Donnelly, R. Newton, G. E. Kennedy, P. S. Fenwick, R. H. F. Leung, K. Ito, R. E. K. Russell, and P. J. Barnes Anti-inflammatory effects of resveratrol in lung epithelial cells: molecular mechanisms Am J Physiol Lung Cell Mol Physiol, October 1, 2004; 287(4): L774 - L783. [Abstract] [Full Text] [PDF]

				J. Leiro, E. Cano, F. M. Ubeira, F. Orallo, and M. L. Sanmartin In Vitro Effects of Resveratrol on the Viability and Infectivity of the Microsporidian Encephalitozoon cuniculi Antimicrob. Agents Chemother., July 1, 2004; 48(7): 2497 - 2501. [Abstract] [Full Text] [PDF]

				S.-Y. Sun, N. Hail Jr, and R. Lotan Apoptosis as a Novel Target for Cancer Chemoprevention J Natl Cancer Inst, May 5, 2004; 96(9): 662 - 672. [Abstract] [Full Text] [PDF]

				N. Osakabe, A. Yasuda, M. Natsume, and T. Yoshikawa Rosmarinic acid inhibits epidermal inflammatory responses: anticarcinogenic effect of Perilla frutescens extract in the murine two-stage skin model Carcinogenesis, April 1, 2004; 25(4): 549 - 557. [Abstract] [Full Text] [PDF]

				A. W. Opipari Jr., L. Tan, A. E. Boitano, D. R. Sorenson, A. Aurora, and J. R. Liu Resveratrol-induced Autophagocytosis in Ovarian Cancer Cells Cancer Res., January 15, 2004; 64(2): 696 - 703. [Abstract] [Full Text] [PDF]

				S. PERVAIZ Resveratrol: from grapevines to mammalian biology FASEB J, November 1, 2003; 17(14): 1975 - 1985. [Full Text] [PDF]

				A. J. Gescher and W. P. Steward Relationship between Mechanisms, Bioavailibility, and Preclinical Chemopreventive Efficacy of Resveratrol: A Conundrum Cancer Epidemiol. Biomarkers Prev., October 1, 2003; 12(10): 953 - 957. [Full Text] [PDF]

				Z. Estrov, S. Shishodia, S. Faderl, D. Harris, Q. Van, H. M. Kantarjian, M. Talpaz, and B. B. Aggarwal Resveratrol blocks interleukin-1{beta}-induced activation of the nuclear transcription factor NF-{kappa}B, inhibits proliferation, causes S-phase arrest, and induces apoptosis of acute myeloid leukemia cells Blood, August 1, 2003; 102(3): 987 - 995. [Abstract] [Full Text] [PDF]

				J. R. Stewart, M. C. Artime, and C. A. O'Brian Resveratrol: A Candidate Nutritional Substance for Prostate Cancer Prevention J. Nutr., July 1, 2003; 133(7): 2440S - 2443. [Abstract] [Full Text] [PDF]

				F. D. Ragione, V. Cucciolla, V. Criniti, S. Indaco, A. Borriello, and V. Zappia p21Cip1 Gene Expression Is Modulated by Egr1: A NOVEL REGULATORY MECHANISM INVOLVED IN THE RESVERATROL ANTIPROLIFERATIVE EFFECT J. Biol. Chem., June 20, 2003; 278(26): 23360 - 23368. [Abstract] [Full Text] [PDF]

				S. C. Pingle, J. F. Sanchez, D. M. Hallam, A. L. Williamson, S. B. Maggirwar, and V. Ramkumar Hypertonicity Inhibits Lipopolysaccharide-Induced Nitric Oxide Synthase Expression in Smooth Muscle Cells by Inhibiting Nuclear Factor {kappa}B Mol. Pharmacol., June 1, 2003; 63(6): 1238 - 1247. [Abstract] [Full Text] [PDF]

				Y. Bayon, M. A. Ortiz, F. J. Lopez-Hernandez, F. Gao, M. Karin, M. Pfahl, and F. J. Piedrafita Inhibition of I{kappa}B Kinase by a New Class of Retinoid-Related Anticancer Agents That Induce Apoptosis Mol. Cell. Biol., February 1, 2003; 23(3): 1061 - 1074. [Abstract] [Full Text]

				K. Ashikawa, S. Majumdar, S. Banerjee, A. C. Bharti, S. Shishodia, and B. B. Aggarwal Piceatannol Inhibits TNF-Induced NF-{kappa}B Activation and NF-{kappa}B-Mediated Gene Expression Through Suppression of I{kappa}B{alpha} Kinase and p65 Phosphorylation J. Immunol., December 1, 2002; 169(11): 6490 - 6497. [Abstract] [Full Text] [PDF]

				J. L. Evans, I. D. Goldfine, B. A. Maddux, and G. M. Grodsky Oxidative Stress and Stress-Activated Signaling Pathways: A Unifying Hypothesis of Type 2 Diabetes Endocr. Rev., October 1, 2002; 23(5): 599 - 622. [Abstract] [Full Text] [PDF]

				U. G. B. Haider, D. Sorescu, K. K. Griendling, A. M. Vollmar, and V. M. Dirsch Resveratrol Suppresses Angiotensin II-Induced Akt/Protein Kinase B and p70 S6 Kinase Phosphorylation and Subsequent Hypertrophy in Rat Aortic Smooth Muscle Cells Mol. Pharmacol., October 1, 2002; 62(4): 772 - 777. [Abstract] [Full Text] [PDF]

				A. V. Gasparian, Y. J. Yao, J. Lu, A. Y. Yemelyanov, L. A. Lyakh, T. J. Slaga, and I. V. Budunova Selenium Compounds Inhibit I{kappa}B Kinase (IKK) and Nuclear Factor-{kappa}B (NF-{kappa}B) in Prostate Cancer Cells Mol. Cancer Ther., October 1, 2002; 1(12): 1079 - 1087. [Abstract] [Full Text] [PDF]

				S. Banerjee, C. Bueso-Ramos, and B. B. Aggarwal Suppression of 7,12-Dimethylbenz(a)anthracene-induced Mammary Carcinogenesis in Rats by Resveratrol: Role of Nuclear Factor-{kappa}B, Cyclooxygenase 2, and Matrix Metalloprotease 9 Cancer Res., September 1, 2002; 62(17): 4945 - 4954. [Abstract] [Full Text] [PDF]

				H. Ferry-Dumazet, O. Garnier, M. Mamani-Matsuda, J. Vercauteren, F. Belloc, C. Billiard, M. Dupouy, D. Thiolat, J. P. Kolb, G. Marit, et al. Resveratrol inhibits the growth and induces the apoptosis of both normal and leukemic hematopoietic cells Carcinogenesis, August 1, 2002; 23(8): 1327 - 1333. [Abstract] [Full Text] [PDF]

				Y. M. Kim, J. Yun, C.-K. Lee, H. Lee, K. R. Min, and Y. Kim Oxyresveratrol and Hydroxystilbene Compounds. INHIBITORY EFFECT ON TYROSINASE AND MECHANISM OF ACTION J. Biol. Chem., May 3, 2002; 277(18): 16340 - 16344. [Abstract] [Full Text] [PDF]

				F. D'Acquisto, M. J. May, and S. Ghosh Inhibition of Nuclear Factor Kappa B (NF-B):: An Emerging Theme in Anti-Inflammatory Therapies Mol. Interv., February 1, 2002; 2(1): 22 - 35. [Abstract] [Full Text] [PDF]

				S. O. SHAHEEN, J. A. C. STERNE, R. L. THOMPSON, C. E. SONGHURST, B. M. MARGETTS, and P. G. J. BURNEY Dietary Antioxidants and Asthma in Adults . Population-based Case-Control Study Am. J. Respir. Crit. Care Med., November 15, 2001; 164(10): 1823 - 1828. [Abstract] [Full Text] [PDF]

				J. Gusman, H. Malonne, and G. Atassi A reappraisal of the potential chemopreventive and chemotherapeutic properties of resveratrol Carcinogenesis, August 1, 2001; 22(8): 1111 - 1117. [Abstract] [Full Text] [PDF]

				K. P. L. Bhat and J. M. Pezzuto Resveratrol Exhibits Cytostatic and Antiestrogenic Properties with Human Endometrial Adenocarcinoma (Ishikawa) Cells Cancer Res., August 1, 2001; 61(16): 6137 - 6144. [Abstract] [Full Text] [PDF]

				N. Ahmad, V. M. Adhami, F. Afaq, D. K. Feyes, and H. Mukhtar Resveratrol Causes WAF-1/p21-mediated G1-phase Arrest of Cell Cycle and Induction of Apoptosis in Human Epidermoid Carcinoma A431 Cells Clin. Cancer Res., May 1, 2001; 7(5): 1466 - 1473. [Abstract] [Full Text]

				M. Q. Holmes-McNary, A. S. Baldwin Jr., and S. H. Zeisel Opposing Regulation of Choline Deficiency-induced Apoptosis by p53 and Nuclear Factor kappa B J. Biol. Chem., October 26, 2001; 276(44): 41197 - 41204. [Abstract] [Full Text] [PDF]

				E. Heiss, C. Herhaus, K. Klimo, H. Bartsch, and C. Gerhauser Nuclear Factor kappa B Is a Molecular Target for Sulforaphane-mediated Anti-inflammatory Mechanisms J. Biol. Chem., August 17, 2001; 276(34): 32008 - 32015. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Holmes-McNary, M. Articles by Baldwin, A. S. Search for Related Content PubMed PubMed Citation Articles by Holmes-McNary, M. Articles by Baldwin, A. S., Jr.