Elsevier

Free Radical Biology and Medicine

Volume 41, Issue 12, 15 December 2006, Pages 1807-1816
Free Radical Biology and Medicine

Original Contribution
DNA damage induced by resveratrol and its synthetic analogues in the presence of Cu (II) ions: Mechanism and structure-activity relationship

https://doi.org/10.1016/j.freeradbiomed.2006.09.007Get rights and content

Abstract

The prooxidant effect of resveratrol (3,5,4′-trihydroxy-trans-stibene) and its synthetic analogues (ArOH), that is, 3,4,4′-trihydroxy-trans-stibene (3,4,4′-THS), 3,4,5-trihydroxy-trans-stibene (3,4,5-THS), 3,4-dihydroxy-trans-stibene (3,4-DHS), 4,4′-dihydroxy-trans-stibene (4,4′-DHS), 2,4-dihydroxy-trans-stilbene (2,4-DHS), 3,5-dihydroxy-trans-stilbene (3,5-DHS) and 3,5,4′-trimethoxy-trans-stibene (3,5,4′-TMS), on supercoiled pBR322 plasmid DNA strand breakage and calf thymus DNA damage in the presence of Cu (II) ions has been studied. It was found that the compounds bearing ortho-dihydroxyl groups (3,4-DHS, 3,4,4′-THS, and 3,4,5-THS) or bearing 4-hydroxyl groups (2,4-DHS, 4,4′-DHS, and resveratrol) exhibit remarkably higher activity in the DNA damage than the ones bearing no such functionalities. Kinetic analysis by UV-visible spectra demonstrates that the formation of ArOH-Cu (II) complexes, the stabilization of oxidative intermediate derived from ArOH and Cu (II)/Cu (I) redox cycles, might be responsible for the DNA damage. This study also reveals a good correlation between antioxidant and prooxidant activity, as well as cytotoxicity against human leukemia (HL-60 and Jurkat) cell lines. The mechanisms and implications of these observations are discussed.

Introduction

Resveratrol (3,5,4′-trihydroxy-trans-stilbene) is a naturally occurring phytoalexin present in grapes, nuts, and other plants. It is believed that the high level of this compound in red wine (0.1–15 mg/L) [1] is linked to the low incidence of heart diseases in some regions of France, the so-called “French paradox;” i.e., despite high fat intake, mortality from coronary heart disease is lower due to the regular drinking of wine [2], [3]. In addition, resveratrol has been shown to possess cancer chemopreventive activity [4], [5], [6], [7]. The metabolism and bioavailability of resveratrol have been studied in detail recently [8]. Therefore, the past several years have witnessed intense research devoted to the biological activity, especially the antioxidative activity, of this compound [9], [10], [11], [12], since free radical-induced oxidative damage of cell membranes, DNA, and protein is considered to play a causative role in aging and several degenerative diseases, such as cancer and atherosclerosis [13], [14], [15], [16]. On the other hand, it was reported that an antioxidant might become a prooxidant to accelerate lipid peroxidation and/or induce DNA damage under special conditions [17], [18], [19], [20], [21], [22], [23], [24], [25], [26], [27]. α-Tocopherol (vitamin E), a well-known antioxidant, was reported to be able to accelerate low-density lipoprotein (LDL) peroxidation via the tocopherol-mediated peroxidation (TMP) [17], [18], and to induce DNA damage in the presence of cupric ions [19]. Other polyphenolic antioxidants, including quercetin [20], curcumin [21], tea cathechins [22], salsolinol [23], and resveratrol [24], [25], [26], [27], were also reported to induce lipid peroxidation and/or DNA damage in the presence of cupric ions. Therefore, it is of interest to see how an antioxidant can switch to a prooxidant and its biological implication.

We found recently that some synthetic resveratrol analogues bearing ortho-diphenoxyl functionality exhibit significantly higher antioxidant and cytotoxic activity against HL-60 cancer cells than resveratrol and other analogues bearing no such functionality [28], [29], [30], [31]. It was reported very recently that resveratrol and other polyphenols could mobilize endogenous copper in human peripheral lymphocytes, leading to oxidative DNA breakage that might be responsible for their anticancer properties [32]. These observations motivated us to study the Cu (II)-mediated oxidative DNA damage by resveratrol and its analogues, putting emphasis on the structure-activity relationship and the mechanism of the prooxidant activity of these compounds. The compounds (ArOH) studied are 3,5,4′-trihydroxy-trans-stibene (resveratrol), 3,4,4′-trihydroxy-trans-stibene (3,4,4′-THS), 3,4,5-trihydroxy-trans-stibene (3,4,5-THS), 3,4-dihydroxy-trans-stibene (3,4-DHS), 4,4′-dihydroxy-trans-stibene (4,4′-DHS), 2,4-dihydroxy-trans-stilbene (2,4-DHS), 3,5-dihydroxy-trans-stilbene (3,5-DHS), and 3,5,4′-trimethoxy-trans-stibene (3,5,4′-TMS).

Molecular Structure

Section snippets

Materials

Resveratrol and its analogues, i.e., 3,4,4′-THS, 3,4,5-THS, 3,4-DHS, 4,4′-DHS, 2,4-DHS, 3,5-DHS, and 3,4,5-TMS were synthesized with reference to the modified Wittig reaction [33], [34] using diethylbenzylphosphonate, which was prepared from methoxyl-substituted benzyl chloride and triethyl phosphate, reacted with methoxyl substituted benzaldehyde, and followed by removing the methyl protecting group with pyridine hydrochloride. This procedure gave exclusively the trans-isomers with moderate to

Strand breakage of plasmid pBR322 DNA induced by resveratrol and its analogues in the presence of Cu (II)

The destruction of the supercoiled pBR322 DNA and formation of the open circular and linear forms of DNA were used to assess the DNA strand breakage [35]. It was found that neither resveratrol nor Cu (II) alone with concentrations up to 200-300 μM showed appreciable effect on the DNA (Lanes 2 and 3 in Fig. 1, Fig. 2, respectively). Other resveratrol analogues (ArOH) also did not induce the DNA strand breakage in the absence of Cu (II) (Fig. 3). However, Cu (II) could work cooperatively with

Discussion

The resveratrol–Cu (II)-induced oxidative DNA damage has been studied by many investigators previously [9], [24], [25], [26], [27], [35]. However, the structural basis for its activity and the relationship between the antioxidant and the prooxidant activity, as well as cytotoxicity of this molecule, have scarcely been exploited [10], [27]. The present work studied the prooxidant activity of eight resveratrol analogues (ArOH) with three different structural features that enables us to deduce a

Acknowledgments

We thank the National Natural Science Foundation of China (Grant Nos. 20502010, 20332020, and 20021001) for financial support.

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