Resveratrol, an antioxidant present in red wine, induces apoptosis in human promyelocytic leukemia (HL-60) cells

https://doi.org/10.1016/S0304-3835(99)00039-7Get rights and content

Abstract

Resveratrol, a triphenolic stilbene present in grapes and other plants, has striking antioxidant and anti-inflammatory activities which have been considered to be responsible for the beneficial effects of red wine consumption on coronary heart disease. Recent studies reveal that resveratrol can inhibit each step of multistage carcinogenesis. However, the molecular mechanisms underlying anti-tumorigenic or chemopreventive activities of this phytochemical remain largely unknown. In the present work, we have found that resveratrol reduces viability and DNA synthesis capability of cultured human promyelocytic leukemia (HL-60) cells. The growth inhibitory and antiproliferative properties of resveratrol appear to be attributable to its induction of apoptotic cell death as determined by morphological and ultrastructural changes, internucleosomal DNA fragmentation, and increased proportion of the subdiploid cell population. Resveratrol treatment resulted in a gradual decrease in the expression of anti-apoptotic Bcl-2. These results, together with previous findings, suggest the cancer therapeutic as well as chemopreventive potential of resveratrol.

Introduction

A wide variety of naturally occurring phenolic substances have been reported to possess substantial anti-tumorigenic activities [1], [2]. Resveratrol (3,5,4′-trihydroxy-trans-stilbene), a phytoalexin present in grapes and other food products, has been suggested as a potential cancer chemopreventive agent based on its striking inhibitory effects on diverse cellular events associated with tumor initiation, promotion, and progression [3]. The compound has strong antioxidative and anti-inflammatory activities which may contribute to its chemopreventive/chemoprotective properties [3], [4], [5], [6]. Recent work by Uenobe and colleagues [7] demonstrated an antimutagenic activity of resveratrol against the food-borne heterocyclic amine, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) in bacteria. Resveratrol attenuated PC12 cell death induced by reactive oxygen species [8]. Synthetic resveratrol has also been found to inhibit growth of human breast epithelial cells in culture [9] and also proliferation of K-562 human erythroleukemia and P-815 murine mastocytoma cells, which may be associated with suppression of ribonucleotide reductase [10].

Recently, considerable attention has focused on the sequence of events referred to as programmed cell death or apoptosis and the role that this process may play in the pathogenesis and treatment of various human diseases including cancer [11]. The maintenance of homeostasis in normal mammalian tissues reflects a critical balance between cell proliferation and cell death via apoptosis. In contrast, apoptosis may be inhibited or perturbed in tumors in which the rate of cell proliferation exceeds that of cell loss. If misregulation of apoptosis results in a failure of tissue size regulation, which eventually leads to the malignant transformation, apoptotic cell death could be induced to augment interventions designed to suppress or reverse the development of cancer. Indeed a variety of cytostatic/cytotoxic drugs have been reported to induce apoptosis in malignant cells in vitro [12], [13]. It is hence conceivable that dietary and/or pharmacological manipulation of apoptosis may provide efficient and promising treatment strategies to protect against cancer [14]. The present study was designed to investigate the possibility that resveratrol could suppress cancer cell growth by inducing apoptotic death.

Section snippets

Chemicals

Resveratrol (∼99% pure), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), sodium dodecylsulfate (SDS), and 4′,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma Chemical Co. (St. Louis, MO). [methyl-3H]Thymidine was obtained from Amersham (Arlington Heights, IL).

Cell culture

Human promyelocytic leukemia (HL-60) cells were obtained from American Type Culture Collection (Rockville, MD) and maintained at 37°C in a humidified atmosphere of 95% air and 5% CO2 in RPMI-1640 (Celox,

Results

Treatment of HL-60 cells with resveratrol was accompanied by marked growth inhibition which was concentration dependent (Fig. 1, upper panel). The addition of resveratrol at 50 μM for 8 h caused about 45% reduction in the cell viability as determined by the conventional MTT assay. The effects of resveratrol on cell proliferation were assessed by measuring DNA synthesis using a radiolabeled precursor. In parallel with the suppression of cell viability, cell proliferation assessed in terms of

Discussion

Chemoprevention, which refers to the use of non-toxic chemical substances to inhibit, delay and/or reverse cellular events associated with carcinogenesis, is regarded as a promising alternative strategy to therapy for the management of cancer [18]. A vast variety of naturally occurring substances have been shown to protect against experimental carcinogenesis and it is becoming increasingly evident that certain phytochemicals, particularly those included in our daily diet have marked cancer

Acknowledgements

This work was supported by the Genetic Engineering Research Grant from the Ministry of Education (1998-019-F00073) awarded to Y.-J. Surh.

References (41)

  • G. Del Bino et al.

    The concentration dependent diversity of effects of DNA topoisomerase I and II inhibitors on the cell cycle of HL-60 cells

    Exp. Cell Res.

    (1991)
  • T. Hirano et al.

    Potent growth inhibitory activity of a novel Ornithogalum cholestane glycoside on human cells: induction of apoptosis in promyelocytic leukemia HL-60 cells

    Life Sci.

    (1996)
  • M.-V. Clement et al.

    Chemopreventive agent resveratrol, a natural product derived from grapes, triggers CD95 signaling-dependent apoptosis in human tumor cells

    Blood

    (1998)
  • Y. Fukamachi et al.

    Zinc suppresses apoptosis of U937 cells induced by hydrogen peroxide through an increase of the Bcl-2/Bax ratio

    Biochem. Biophys. Res. Commun.

    (1998)
  • M.-T. Huang et al.

    Phenolic compounds in food and cancer prevention

  • Y.-J. Surh

    Cancer chemoprevention by dietary phytochemicals: a mechanistic viewpoint

    Cancer J.

    (1998)
  • M. Jang et al.

    Cancer chemopreventive activity of resveratrol, a natural product derived from grapes

    Science

    (1997)
  • K. Pattichis et al.

    Inhibition of human LDL oxidation by resveratrol

    Lancet

    (1993)
  • D.M. Goldberg

    More on antioxidant activity of resveratrol in red wine

    Clin. Chem.

    (1996)
  • S. Chanvitayapongs et al.

    Amelioration of oxidative stress by antioxidants and resveratrol in PC12 cells

    NeuroReport

    (1997)
  • Cited by (0)

    View full text