The aim of this work was to evaluate in vitro the genotoxic and/or antigenotoxic effects of resveratrol (RESV) and pterostilbene (PTER) on HepG2 cells. Moreover, additional tests were performed to evaluate early and late apoptosis events induced by the tested stilbenes. RESV and PTER did not show any genotoxic activity. As regards antigenotoxicity testing, RESV and PTER showed a typical, U-shaped hormetic dose-response relationship characterized by a biphasic trend with small quantities having opposite effects to large ones. HepG2 cells treated with PTER exhibited a marked increase in early apoptosis (40.1 %) at 250 μM; whereas, the highest concentration tested for both RESV and PTER significantly increased the proportion of HepG2 cells undergoing late apoptosis (32.5 and 51.2 %, respectively). The observed pro-apoptotic activity could, at least in part, explain the hormetic response observed when the compounds were tested for antigenotoxicity (i.e., in the presence of induced DNA damage).
MahadyGB, PendlandSL, ChadwickLR. (2003) Resveratrol and red wine extracts inhibit the growth of CagA+ strains of Helicobacter pylori in vitro. The American Journal of Gastroenterology, 98, 1440–1441.
2.
RimandoAM, SuhN. (2008) Biological/chemopreventive activity of stilbenes and their effect on colon cancer. Planta Medica, 74, 1635–1643.
3.
ValenzanoDR, TerzibasiE, GenadeT, CattaneoA, DomeniciL, CellerinoA. (2006) Resveratrol prolongs lifespan and retards the onset of age-related markers in a short-lived vertebrate. Current Biology, 16, 296–300.
4.
WangY, DingL, WangX, ZhangJ, HanW, FengL, SunJ, JinH, WangXJ. (2012) Pterostilbene simultaneously induces apoptosis, cell cycle arrest and cyto-protective autophagy in breast cancer cells. American Journal of Translational Research, 4, 44–51.
5.
AbrahamSK, EckhardtA, OliRG, StopperH. (2012) Analysis of in vitro chemoprevention of genotoxic damage by phytochemicals, as single agents or as combinations. Mutation Research, 744, 117–124.
6.
SgambatoA, ArditoR, FaragliaB, BoninsegnaA, WolfFI, CittadiniA. (2001) Resveratrol, a natural phenolic compound, inhibits cell proliferation and prevents oxidative DNA damage. Mutation Research, 496, 171–180.
7.
PanMH, ChangYH, BadmaevV, NagabhushanamK, HoCT. (2007) Pterostilbene induces apoptosis and cell cycle arrest in human gastric carcinoma cells. Journal of Agricultural and Food Chemistry, 55, 7777–7785.
8.
AzmiAS, BhatSH, HanifS, HadiSM. (2006) Plant polyphenols mobilize endogenous copper in human peripheral lymphocytes leading to oxidative DNA breakage: a putative mechanism for anticancer properties. FEBS Letters, 580, 533–538.
9.
ChangJ, RimandoA, PallasM, CaminsA, PorquetD, ReevesJ, Shukitt-HaleB, SmithMA, JosephJA, CasadesusG. (2012) Low-dose pterostilbene, but not resveratrol, is a potent neuromodulator in aging and Alzheimer's disease. Neurobiology of Aging, 33, 2062–2071.
10.
ChenRJ, HoCT, WangYJ. (2010) Pterostilbene induces autophagy and apoptosis in sensitive and chemoresistant human bladder cancer cells. Molecular Nutrition & Food Research, 54, 1819–1832.
11.
KunduJK, SurhYJ. (2008) Cancer chemopreventive and therapeutic potential of resveratrol: mechanistic perspectives. Cancer Letters, 269, 243–261.
12.
MannalP, McDonaldD, McFaddenD. (2010) Pterostilbene and tamoxifen show an additive effect against breast cancer in vitro. American Journal of Surgery, 200, 577–580.
13.
McCormackD, SchneiderJ, McDonaldD, McFaddenD. (2011) The antiproliferative effects of pterostilbene on breast cancer in vitro are via inhibition of constitutive and leptin-induced Janus kinase/signal transducer and activator of transcription activation. American Journal of Surgery, 202, 541–544.
14.
PatelKR, ScottE, BrownVA, GescherAJ, StewardWP, BrownK. (2011) Clinical trials of resveratrol. Annals of the New York Academy of Sciences, 1215, 161–169.
15.
RoupeKA, RemsbergCM, YanezJA, DaviesNM. (2006) Pharmacometrics of stilbenes: seguing towards the clinic. Current Clinical Pharmacology, 1, 81–101.
16.
LangcakeP, CornfordC, PryceR. (1979) Identification of pterostilbene as a phytoalexin from Vitis vinifera leaves. Phytochemistry, 18, 1025–1027.
17.
LangcakeP, PryceR. (1976) The production of resveratrol by Vitis vinifera and other members of the Vitaceae as a response to infection or injury. Physiological Plant Pathology, 9, 77–86.
18.
FerrerP, AsensiM, SegarraR, OrtegaA, BenllochM, ObradorE, VareaMT, AsensioG, JordaL, EstrelaJM. (2005) Association between pterostilbene and quercetin inhibits metastatic activity of B16 melanoma. Neoplasia, 7, 37–47.
19.
LinHS, YueBD, HoPC. (2009) Determination of pterostilbene in rat plasma by a simple HPLC-UV method and its application in pre-clinical pharmacokinetic study. Biomedical Chromatography: BMC, 23, 1308–1315.
20.
McCormackD, McFaddenD. (2012) Pterostilbene and cancer: current review. The Journal of Surgical Research, 173, e53–61.
21.
PanZ, AgarwalAK, XuT, FengQ, BaersonSR, DukeSO, RimandoAM. (2008) Identification of molecular pathways affected by pterostilbene, a natural dimethylether analog of resveratrol. BMC Medical Genomics, 1, 7 [on line].
22.
TiceRR, AgurellE, AndersonD, BurlinsonB, HartmannA, KobayashiH, MiyamaeY, RojasE, RyuJC, SasakiYF. (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environmental and Molecular Mutagenesis, 35, 206–221.
23.
SalvioliS, ArdizzoniA, FranceschiC, CossarizzaA. (1997) JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess delta psi changes in intact cells: implications for studies on mitochondrial functionality during apoptosis. FEBS Letters, 411, 77–82.
24.
NagataS. (2000) Apoptotic DNA fragmentation. Experimental Cell Research, 256, 12–18.
25.
NagataS, NagaseH, KawaneK, MukaeN, FukuyamaH. (2003) Degradation of chromosomal DNA during apoptosis. Cell Death and Differentiation, 10, 108–116.
26.
HayesDP. (2008) Adverse effects of nutritional inadequacy and excess: a hormetic model. The American Journal of Clinical Nutrition, 88, 578S–581S.
27.
CalabreseEJ, MattsonMP, CalabreseV. (2010) Resveratrol commonly displays hormesis: occurrence and biomedical significance. Human & Experimental Toxicology, 29, 980–1015.
28.
MukherjeeS, DudleyJI, DasDK. (2010) Dose-dependency of resveratrol in providing health benefits. Dose-Response: A Publication of International Hormesis Society, 8, 478–500.
DarzynkiewiczZ, JuanG, LiX, GorczycaW, MurakamiT, TraganosF. (1997) Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis). Cytometry, 27, 1–20.
31.
HanX, WangF, YaoW, XingH, WengD, SongX, ChenG, XiL, ZhuT, ZhouJ, XuG, WangS, MengL, IadecolaC, WangG, MaD. (2007) Heat shock proteins and p53 play a critical role in K+ channel-mediated tumor cell proliferation and apoptosis. Apoptosis, 12, 1837–1846.
32.
ParkerBW, KaurG, Nieves-NeiraW, TaimiM, KohlhagenG, ShimizuT, LosiewiczMD, PommierY, SausvilleEA, SenderowiczAM. (1998) Early induction of apoptosis in hematopoietic cell lines after exposure to flavopiridol. Blood, 91, 458–465.
33.
LeeI, ChoeYS, ChoiJY, LeeKH, KimBT. (2012) Synthesis and evaluation of (1)(8)F-labeled styryltriazole and resveratrol derivatives for beta-amyloid plaque imaging. Journal of Medicinal Chemistry, 55, 883–892.
34.
DiamondL, KruszewskiF, AdenDP, KnowlesBB, BairdWM. (1980) Metabolic activation of benzo[a]pyrene by a human hepatoma cell line. Carcinogenesis, 1, 871–875.
35.
SassaS, SugitaO, GalbraithRA, KappasA. (1987) Drug metabolism by the human hepatoma cell, Hep G2. Biochemical and Biophysical Research Communications, 143, 52–57.
36.
KnasmullerS, ParzefallW, SanyalR, EckerS, SchwabC, UhlM, Mersch-SundermannV, WilliamsonG, HietschG, LangerT, DarroudiF, NatarajanAT. (1998) Use of metabolically competent human hepatoma cells for the detection of mutagens and antimutagens. Mutation Research, 402, 185–202.
37.
ShahD, NaciriM, CleeP, Al-RubeaiM. (2006) NucleoCounter-An efficient technique for the determination of cell number and viability in animal cell culture processes. Cytotechnology, 51, 39–44.
38.
SinghNP, McCoyMT, TiceRR, SchneiderEL. (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research, 175, 184–191.
CollinsAR. (2004) The comet assay for DNA damage and repair: principles, applications, and limitations. Molecular Biotechnology, 26, 249–261.
41.
LovellDP, OmoriT. (2008) Statistical issues in the use of the comet assay. Mutagenesis, 23, 171–182.
42.
VillariniM, PagiottiR, DominiciL, FatigoniC, VanniniS, LevoratoS, MorettiM. (2014) Investigation of the cytotoxic, genotoxic, and apoptosis-inducing effects of estragole isolated from fennel (Foeniculum vulgare). Journal of Natural Products, 77, 773–778.
43.
KajsturaM, HalickaHD, PryjmaJ, DarzynkiewiczZ. (2007) Discontinuous fragmentation of nuclear DNA during apoptosis revealed by discrete “sub-G1” peaks on DNA content histograms. Cytometry. Part A: The Journal of the International Society for Analytical Cytology, 71, 125–131.
