Purpose: To investigate the effect and possible mechanisms of resveratrol on pancreatic cancer cells in vitro. Methods: After being treated with resveratrol, cell viability, cell cycle phase distribution and apoptosis rate of pancreatic cancer cells were measured by CCK-8 assay and flow cytometer, respectively. The effects of resveratrol on the Hedgehog pathway were studied by real-time RT-PCR and Western blotting. By interfering Gli1 expression in PANC-1 cells and overexpressing Gli1 in BxPC-3 cells, we detected the expressions of Gli1-targeted genes, such as Ptc1, CCND1 and BCL-2, compared with resveratrol experimental group. We further used the luciferase reporter assay to explore the correlation between resveratrol and Gli1. Results: Resveratrol inhibited the growth of pancreatic cancer cells in a dose- and time-dependent manner. Compared with control group, the cells in the G0/G1 phase and the apoptosis rate were significantly increased. Low concentration of resveratrol decreased the expression of the Hedgehog pathway members including Gli1, Ptc1 and Smo. The expression of downstream target genes of the Hedgehog pathway such as Gli1, Ptc1, CCND1 and BCL-2 were significantly decreased after 12.5 µM resveratrol treatment, which demonstrated a similar change of gene expression when Gli1 was knocked down by the RNAi technique in PANC-1 cells. Resveratrol also downregulated the expression of Gli1, Ptc1, CCND1 and BCL-2 in Gli1-overexpressed BxPC-3 cells. Results of the luciferase assay showed that resveratrol did not act on the Gli1 promoter directly. Conclusion: Resveratrol can inhibit pancreatic cancer cell survival and its mechanisms might be partly via the Hedgehog signaling pathway.

Keywords:
Resveratrol, Pancreatic cancer, Hedgehog signaling pathway, Proliferation, Apoptosis
1.
Kayahara M, Funaki K, Tajima H, Takamura H, Ninomiya I, Kitagawa H, Ohta T: Surgical implication of micrometastasis for pancreatic cancer. Pancreas 2010;39:884–888.
2.
Mayo SC, Austin DF, Sheppard BC, Mori M, Shipley DK, Billingsley KG: Adjuvant therapy and survival after resection of pancreatic adenocarcinoma: a population-based analysis. Cancer 2010;116:2932–2940.
3.
Richter J, Saif MW: Updates in adjuvant therapy in pancreatic cancer: gemcitabine and beyond. Highlights from the ‘2010 ASCO Gastrointestinal Cancers Symposium’, Orlando, USA. JOP 2010;11:144–147.
4.
Li D, Xie K, Wolff R, Abbruzzese JL: Pancreatic cancer. Lancet 2004;363:1049–1057.
5.
Hebrok M: Hedgehog signaling in pancreas development. Mech Dev 2003;120:45–57.
6.
Caro I, Low JA: The role of the Hedgehog signaling pathway in the development of basal cell carcinoma and opportunities for treatment. Clin Cancer Res 2010;16:3335–3339.
7.
Barginear MF, Leung M, Budman DR: The Hedgehog pathway as a therapeutic target for treatment of breast cancer. Breast Cancer Res Treat 2009;116:239–246.
8.
Saqui-Salces M, Merchant JL: Hedgehog signaling and gastrointestinal cancer. Biochim Biophys Acta 2010;1803:786–795.
9.
Thayer SP, di Magliano MP, Heiser PW, Nielsen CM, Roberts DJ, Lauwers GY, Qi YP, Gysin S, Fernández-del Castillo C, Yajnik V, Antoniu B, McMahon M, Warshaw AL, Hebrok M: Hedgehog is an early and late mediator of pancreatic cancer tumorigenesis. Nature 2003;425:851–856.
10.
Jones S, Zhang X, Parsons DW, et al: Core signaling pathways in human pancreatic cancers revealed by global genomic analyses. Science 2008;321:1801–1806.
11.
Feldmann G, Habbe N, Dhara S, Bisht S, Alvarez H, Fendrich V, Beaty R, Mullendore M, Karikari C, Bardeesy N, Ouellette MM, Yu W, Maitra A: Hedgehog inhibition prolongs survival in a genetically engineered mouse model of pancreatic cancer. Gut 2008;57:1420–1430.
12.
Lee CJ, Dosch J, Simeone DM: Pancreatic cancer stem cells. J Clin Oncol 2008;6:2806–2812.
13.
Mueller MT, Hermann PC, Witthauer J, Rubio-Viqueira B, Leicht SF, Huber S, Ellwart JW, Mustafa M, Bartenstein P, D’Haese JG, Schoenberg MH, Berger F, Jauch KW, Hidalgo M, Heeschen C: Combined targeted treatment to eliminate tumorigenic cancer stem cells in human pancreatic cancer. Gastroenterology 2009;137:1102–1113.
14.
Jimeno A, Feldmann G, Suárez-Gauthier A, Rasheed Z, Solomon A, Zou GM, Rubio-Viqueira B, García-García E, López-Ríos F, Matsui W, Maitra A, Hidalgo M: A direct pancreatic cancer xenograft model as a platform for cancer stem cell therapeutic development. Mol Cancer Ther 2009;8:310–314.
15.
Pervaiz S, Holme AL: Resveratrol: its biologic targets and functional activity. Antioxid Redox Signal 2009;11:2851–2897.
16.
Jang M, Cai L, Udeani GO, et al: Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 1997;275:218–220.
17.
Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CW, Fong HH, Farnsworth NR, Kinghorn AD, Mehta RG, Moon RC, Pezzuto JM: Antiproliferative effect of resveratrol in pancreatic cancer cells. Phytother Res 2010;24:1637–1644.
18.
Oi N, Jeong CH, Nadas J, Cho YY, Pugliese A, Bode AM, Dong Z: Resveratrol, a red wine polyphenol, suppresses pancreatic cancer by inhibiting leukotriene a4 hydrolase. Cancer Res 2010;70:9755–9764.
19.
Harikumar KB, Kunnumakkara AB, Sethi G, Diagaradjane P, Anand P, Pandey MK, Gelovani J, Krishnan S, Guha S, Aggarwal BB: Resveratrol, a multitargeted agent, can enhance antitumor activity of gemcitabine in vitro and in orthotopic mouse model of human pancreatic cancer. Int J Cancer 2010;127:257–268.
20.
Wang F, Xu L, Guo C, Ke A, Hu G, Xu X, Mo W, Yang L, Huang Y, He S, Wang X: Identification of RegIV as a novel GLI1 target gene in human pancreatic cancer. PloS One 2011;11:e18434.
21.
Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ: Cancer Statistics, 2009. CA Cancer J Clin 2009;59:225–249.
22.
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;24:2783–2840.
23.
Athar M, Back JH, Kopelovich L, Bickers DR, Kim AL: Multiple molecular targets of resveratrol: Anti-carcinogenic mechanisms. Arch Biochem Biophys 2009;486:95–102.
24.
Delmas D, Solary E, Latruffe N: Resveratrol, a phytochemical inducer of multiple cell death pathways: apoptosis, autophagy and mitotic catastrophe. Curr Med Chem 2011;18:1100–1121.
25.
Shankar S, Nall D, Tang SN, Meeker D, Passarini J, Sharma J, Srivastava RK: Resveratrol inhibits pancreatic cancer stem cell characteristics in human and KrasG12D transgenic mice by inhibiting pluripotency maintaining factors and epithelial-mesenchymal transition. PLoS One 2011;6:e16530.
26.
Tili E, Michaille JJ: Resveratrol, MicroRNAs, inflammation, and cancer. J Nucl Acids 2011;2011:102431.
27.
Araújo JR, Gonçalves P, Martel F; Chemopreventive effect of dietary polyphenols in colorectal cancer cell lines. Nutr Res 2011;31:77–87.
28.
Xu FG, Ma QY, Wang Z: Blockade of Hedgehog signaling pathway as a therapeutic strategy for pancreatic cancer. Cancer Lett 2009;283:119–124.
29.
Takaori K: Current understanding of precursors to pancreatic cancer. J Hepatobiliary Pancreat Surg 2007;14:217–223.
30.
Katoh Y, Katoh M: Hedgehog target genes: mechanisms of carcinogenesis induced by aberrant Hedgehog signaling activation. Curr Mol Med 2009;9:873–886.
31.
Colin D, Limagne E, Jeanningros S, Jacquel A, Lizard G, Athias A, Gambert P, Hichami A, Latruffe N, Solary E, Delmas D: Endocytosis of resveratrol via lipid rafts and activation of downstream signaling pathways in cancer cells. Cancer Prev Res 2011:1095–1106.
32.
Kasper M, Schnidar H, Neill GW, Hanneder M, Klingler S, Blaas L, Schmid C, Hauser-Kronberger C, Regl G, Philpott MP, Aberger F: Selective modulation of Hedgehog/GLI target gene expression by epidermal growth factor signaling in human keratinocytes. Mol Cell Biol 2006;26:6283–6298.
33.
Hegde GV, Peterson KJ, Emanuel K, Mittal AK, Joshi AD, Dickinson JD, Kollessery GJ, Bociek RG, Bierman P, Vose JM, Weisenburger DD, Joshi SS: Hedgehog-induced survival of B-cell chronic lymphocytic leukemia cells in a stromal cell microenvironment: a potential new therapeutic target. Mol Cancer Res 2008;6:1928–1936.
34.
Wang K, Pan L, Che X, Cui D, Li C: Gli1 inhibition induces cell-cycle arrest and enhanced apoptosis in brain glioma cell lines. J Neurooncol 2010;98:319–327.
© 2012 S. Karger AG, Basel and IAP
2012
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.