Cancer Cell & Microenvironment 2014;1:e29. http://www.smartscitech.com/index.php/ccm
EXPERT RESEARCH HIGHLIGHT
Marine bioactive products as anti-cancer agents: effects of sea anemone venom on breast and lung cancer cells 1
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Mahnaz Ramezanpour , Karen Burke da Silva , Barbara J.S. Sanderson
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Department of Medical Biotechnology, School of Medicine, Flinders University of South Australia, Bedford Park, SA 5042, Australia 2 School of Biological Sciences, Flinders University of South Australia GPO, Box 2100, Adelaide 5001, Australia
Correspondence: Barbara J.S. Sanderson E-mail:
[email protected] Received: December 04, 2013 Published online: January 15, 2014 Cancer cell & Microenvironment 2014; 1: e29. doi: 10.14800/ccm.29; © 2014 by Mahnaz Ramezanpour, et al.
phase with an associated accumulation of cells in G0/G1 phase [13]. In contrast, H. magnifica venom applied to human breast cancer T47D and MCF7 cells was found to induce cell cycle arrest in sub G1 phase with a concomitant decrease in the G1 phase, indicating induction of apoptosis [12] . This discrepancy might be due to the difference in cell type analysed. Apoptosis is an important defence against cancer [15]. However, the resistance of human cancer to apoptosis may be due to inactivation of proapoptotic effectors (e.g., loss of the p53 pathway) or overexpression of Bcl-2 family, which can block apoptosis [16, 17]. Therefore, apoptotic pathways are emerging as promising targets for cancer therapy. We used flow cytometry assay for apoptosis, and the experimental data confirmed that venom from H. magnifica could induce apoptosis (Figure 1). However, far fewer papers are published each year on apoptosis in lung cells than in the other major organs. H. magnifica venom is a candidate to induce apoptosis in non-small-cell lung cancer A549 cells [13].
Introduction Marine organisms abound that have a rich heritages as therapeutics resources that have been exploited for effective and beneficial use against many human cancers. Many of these natural compounds in are venom used to acquire prey and as chemical defence against predators in nature [1]. Numerous toxic peptides are found within these venoms, and some of them have been used to investigate biomedical problems including designing novel drugs [2]. Sea anemones are one of the most ancient predatory animals and various in vitro and in vivo studies have demonstrated that more than 32 species of sea anemones studied produce lethal cytolytic peptides and proteins [3]. Many biological assays have been carried out to evaluate toxicity of the venoms. Studies of hemolytic activities [4, 5] immunomodulating activities[6, 7], neurotoxic[8], cardiotox-ic[9] and cytolytic tests [3, 10, 11] have all produced interes-ting outcomes. Some recent observations of the venom from the sea anemone Heteractis magnifica, have shown that it can induce apoptosis in human breast and lung cancer cell lines [12, 13]. Cell cycle deregulation is an important feature of many cancers and therefore control of cell cycle progression is suggested to be an effective strategy to inhibit cancer growth [14]. Clear evidence of cell cycle arrest and apoptosis was observed in our previous study. A549 cancer cells treated with of H. magnifica venom showed delayed progression through the cell cycle and a marked reduction in the number of cells in S
Apoptosis occurs via two principal pathways, the death receptor pathway (extrinsic) and the mitochondrial pathway (intrinsic). The extrinsic pathway is triggered by the binding of death inducing ligands to cell surface receptors, which results in the activation of caspase 8. The intrinsic pathway, in contrast, is triggered by cytotoxic stress, which converges at the mitochondria, leading to the release of cytochrome c from mitochondria. In the cytosol, cytochrome c binds Apaf
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Mahnaz Ramezanpour, et al. Effects of sea anemone venom on breast and lung cancer cells
treatment [25] and also did not express detectable level of caspase 3[26]. H.magnifica venom however, could interfere with the cell cycle at the sub G1 phase, induce apoptosis and increase the activation of caspase 3/7[12]. An increase of mitochondrial membrane permeability is one of the key events in apoptotic pathway by releasing apoptogenic proteins into the cytosol [27-29]. Our previous work demonstrated that H. magnifica venom induced apoptosis by loss of mitochondrial membrane potential of human non-small cell lung cancer A549 cells [13]. These findings are consistent with our previous study on T47D and MCF7 cell lines that the venom significantly increased the percentage of cells positive for JC-1 monomers[12]. Many previous reports have shown that sea anemone venoms exert cytotoxic effects on different cancer cell lines. We have developed the mechanisms action of the H. magnifica venom. H. magnifica venom has great potential to induce significant apoptosis and cell cycle arrest in lung and breast cancer cell lines. Further study could be carried out using more cancer cell line with different background to design of combination treatment using the H. magnifica venom with other therapies, based on molecular targets.
Fig 1. The effect of H. magnifica venom on apoptosis in A549, T47D and MCF7 cell lines as determined by Annexin V-conjugated PI staining followed by flow cytometry. Data from 10,000 events and percentage of early apoptotic cells (Annexin positive) and late apoptotic cells (Annexin positive/PI positive) are shown. Values for each cell line are shown as mean ± SEM from 3 independent experiments, following treatment with 40 µg/ml venom. The control values are the mean ± SEM from all cell lines not treated with venom (n=9).
Acknowledgments This study was supported by a FMC Foundation Grant and a Flinders Centre for Innovation in Cancer (FCIC) research grant. References 1. Banerjee S, Wang Z, Mohammad M, Sarkar FH, Mohammad R. Efficacy of selected natural products as therapeutic agents against cancer. J Nat Prod. 2008;71:492-496. 2. Andreev YA, Kozlov SA, Koshelev SG, Ivanova EA, Monastyrnaya MM, Kozlovskaya EP, et al. Analgesic compound from sea anemone Heteractis crispa is the first polypeptide inhibitor of vanilloid receptor 1 (TRPV1). J Biol Chem. 2008;283:2391423921.
Fig 2. Caspase activity for A549, T47D and MCF7 cell lines after 24 h treatment with venom from H. magnifica, assayed using a luminescent kit. Data are presented as relative luminescence unit (RLU) cells/s (104). Values are shown as mean ± SEM of 3 independent experiments. *Significantly different from the untreated control at p>0.05.
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in the cytoplasm. The Apaf/cytochrome c complex binds caspase 9, to form an active complex called the apoptosome. Activation of caspase 3/7 (effector caspases) is involved in both pathways, while caspase 8 and 9 (initiator caspases) are involved in extrinsic and intrinsic pathway, respectively[15, 18, 19] . Effector caspases, include-ing caspase 3/7, activate DNase resulting in fragmenta-tion of DNA in response to various apoptotic stimuli [20-24]. Recently, we reported that venom from H.magnifica was able to induced cell death by activation of the caspase pathway. H. magnifica venom significantly incr-eased the levels of caspase 3/7, enzyme activity in A549, T47D and MCF7 cell lines (Figure 2) [12, 13] . It has been reported that MCF7 cells do not show an increase in the sub-G1 in response to anti-cancer drug
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To cite this article: Ramezanpour M, et al. Marine bioactive products as anti-cancer agents: effects of sea anemone venom on breast and lung cancer cells. Cancer Cell Microenviron 2014; 1: e29. doi: 10.14800/ccm.29.
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