Antitumor Effects of Zerumbone from Zingiber zerumbet in P-388D₁ Cells in Vitro and in Vivo

Abstract

The fresh rhizome of Zingiber zerumbet (L.) Roscoe ex Smith (Zingiberaceae) is widely used as a folk medicine in Taiwan. In this study, the fresh rhizome was extracted with 95 % EtOH and partitioned with diethyl ether. The antitumor effects of the diethyl ether extract were measured in cultured P-388D₁ cells and in an animal model of P-388D₁-bearing CDF₁ mice. The results indicated that the extract could induce DNA fragmentation in P-388D₁ cells in vitro, and significantly prolong the life of P-388D₁-bearing CDF₁ mice (ILS% = 127.8) at a dosage of 5 mg/kg body weight. After column chromatography combined with an MTT cytotoxicity bioassay, zerumbone, a cyclic sesquiterpene was isolated from the diethyl ether extract. Zerumbone inhibited the growth of P-388D₁ cells, induced DNA fragmentation in culture, and significantly prolonged the life of P-388D₁-bearing CDF₁ mice (ILS% = 120.5) at a dosage of 2 mg/kg. Furthermore, zerumbone inhibited the growth of a human leukemia cell line, HL-60 cells, in a time- and concentration-dependent manner, with IC₅₀ values of 22.29, 9.12, and 2.27 μg/mL for 6, 12, and 18 h, respectively. The cell cycle of HL-60 cells was observed after treatment with zerumbone, which induced G₂/M cell cycle arrest in HL-60 cells in a time- and concentration-dependent manner, and decreased the cyclin B1/cdk 1 protein level. These results suggest that zerumbone is an active principal of Z. zerumbet and is potentially a lead compound for the development of anticancer drugs.

Key words

Zingiber zerumbet - Zingiberaceae - antitumor - zerumbone - P-388D₁ cell - cell cycle

References

• 1 Surh Y. Molecular mechanisms of chemopreventive effects of selected dietary and medicinal phenolic substances.  Mutat Res. 1999;  428 305-27
• 2 Tanaka T, Shimizu M, Kohno H, Yoshitani S, Tsukio Y, Murakami A, Safitri R, Takahashi D, Yamamoto K, Koshimizu K, Ohigashi H, Mori H. Chemoprevention of azoxymethane-induced rat aberrant crypt foci by dietary zerumbone isolated from Zingiber zerumbet .  Life Sci. 2001;  69 1935-45
• 3 Chiu N Y, Chang K H. The illustrated medicinal plants of Taiwan. 1995: 2, 285
• 4 Damodaran N P, Sukh D. Studies in sesquiterpenes-XXX sesquiterpenes from the essential oil of Zingiber zerumbet Smith.  Tetrahedron. 1968;  24 4113-22
• 5 Matthes H WD, Luu B, Ourisson G. Cytotoxic compounds of Zingiber zerumbet, Curcuma zedoaria and C. domestica .  Phytochemistry. 1980;  19 2643-50
• 6 Masuda T, Jitoe A, Kato S, Nobuji N. Acetylated flavonol glycosides from Zingiber zerumbet .  Phytochemistry. 1991;  20 2391-2
• 7 Murakami A, Takahashi M, Jiwajinda S, Koshimizu K, Ohigashi H. Identification of zerumbone in Zingiber zerumbet Smith as a potent inhibitor of 12-O-tetradecanoylphorbol-13-acetate-induced Epstein-Barr virus activation.  Biosci Biotechnol Biochem. 1999;  63 1811-2
• 8 Murakami A, Takahashi D, Kinoshita T, Koshimizu K, Kim H W, Yoshihiro A, Nakamura Y, Jiwajinda S, Terao J, Ohigashi H. Zerumbone, a Southeast Asian ginger sesquiterpene, markedly suppresses free radical generation, proinflammatory protein production, and cancer cell proliferation accompanied by apoptosis: the alpha, beta-unsaturated carbonyl group is a prerequisite.  Carcinogenesis. 2002;  23 795-802
• 9 Chen F C, Liu Y T, Chen H Y, Lee W, Lin H C, Wu C H, King M L. Mouse lymphocytic leukemia P388 for screening antitumor agents from crude plant extracts and fermentation broth.  Chin Pharm J. 1992;  44 381-8
• 10 Gebhardt R. In vitro screening of plant extracts and phytopharmaceuticals: novel approaches for the elucidation of active compounds and their mechanisms.  Planta Med. 2000;  66 99-105
• 11 Kitayama T, Okamoto T, Hill R K, Kawai Y, Takahashi S, Yonemori S, Yamamoto Y, Ohe K, Uemura S, Sawada S. Chemistry of zerumbone. 1. Simplified isolation, conjugate addition reactions, and a unique ring contracting transannular reaction of its dibromide.  J Org Chem. 1999;  64 2667-72
• 12 Bruggisser R, Daeniken K V, Jundt G, Schaffner W, Tullberg-Reinert H. Interference of plant extracts, phytoestrogens and antioxidants with the MTT tetrazolium assay.  Planta Med. 2002;  68 445-8
• 13 Wang C C, Chen L G, Yang L L. Cytotoxic activity of sesquiterpenoids from Atractylodes ovata on leukemia cell lines.  Planta Med. 2002;  68 204-8
• 14 Yang LL,Chang C C, Chen L G, Wang C C. Antitumor principle constituents of Myrica rubra var. acuminata .  J Agric Food Chem. 2003;  51 2974-9
• 15 Hartwell L H, Weinert T A. Checkpoints: controls that ensure the order of cell cycle events.  Science. 1989;  246 629-34
• 16 Malumbres M, Barbacid M. To cycle or not to cycle: a critical decision in cancer.  Nat Rev Cancer. 2001;  1 222-31
• 17 Jackman M, Lindon C, Nigg E A, Pines J. Active cyclin B1-Cdk1 first appears on centrosomes in prophase.  Nat Cell Bill. 2003;  5 143-8
• 18 Murakami A, Takahashi D, Koshimizu K, Ohigashi H. Synergistic suppression of superoxide and nitric oxide generation from inflammatory cells by combined food factors.  Mutat Res. 2003;  523 151-61
• 19 Liao Y H, Houghton P J, Hoult J R. Novel and known constituents from Buddleja species and their activity against leukocyte eicosanoid generation.  J Nat Prod. 1999;  62 1241-5
• 20 Murakami A, Tanaka T, Lee J Y, Surh Y J, Kim H W, Kawabata K, Nakamura Y, Jiwajinda S, Ohigashi H. Zerumbone, a sesquiterpene in subtropical ginger, suppresses skin tumor initiation and promotion stages in ICR mice.  Int J Cancer. 2004;  110 481-90