Cytotoxic and genotoxic effects of tambjamine D, an alkaloid isolated from the nudibranch Tambja eliora, on Chinese hamster lung fibroblasts
Introduction
Marine invertebrates are an important source of bioactive natural products, many of which exhibit unique structural features not found in natural products derived from terrestrial organisms [1], [2]. As a consequence, marine organisms have attracted attention as potential sources of bioactive substances, and they continue to provide novel lead compounds for the pharmaceutical industry [3], [4], [5], [6], [7], [8].
Tambjamines are representative members of the 4-methoxypyrrolic alkaloids derived from marine sources, which exhibit cytotoxic, antimicrobial and immunosuppressive activities [9], [10]. Tambjamines A–D have been isolated from the nudibranchs Tambja eliora, T. abdere and Roboastra tigris and been shown to have antibacterial activity against Escherichia coli, Staphylococcus aureus and Vibrio anguillarum[11], [12]. Tambjamine D isolated from the nudibranch T. eliora was also found to display antibacterial activity against B. subtilis, antifungal activity against Candida albicans, antimitotic effects in the sea urchin egg development assay, and cytotoxic activity against several tumor cell lines [13].
Some of the biological effects of 4-methoxypyrrolic alkaloids may be related to their ability to intercalate DNA [14], [15]. DNA-intercalating agents such as acridines are known to cause several mutagenic events such as DNA strand breaks and frameshift mutations [16], [17]. Melvin et al. [15] also demonstrated that 4-methoxypyrrolic from natural products bind double-stranded DNA and facilitate its oxidative cleavage.
Considering the DNA-targeting properties of these compounds, the aim of the present study was to investigate the cytotoxic and genotoxic effects of the alkaloid tambjamine D isolated from the nudibranch T. eliora using V79 cells a permanent cell line derived from Chinese hamster lung fibroblasts. This study is important for determining the safety of a possible pharmacological application of this compound and for exploring other biological properties. For this purpose, we determined the cytotoxicity of tambjamine D using three different endpoints, and evaluated the oxidative stress promoted by tambjamine D by measuring nitrite/nitrate production and TBARS formation after cell exposure to tambjamine D. In order to obtain a complete evaluation of the toxic effects of this alkaloid in mammalian cells, DNA damage induced by tambjamine D was estimated using the comet assay and the micronucleus test.
Section snippets
Chemicals
Tambjamine D (Fig. 1) was isolated from T. eliora as previously described [13]. Modified Eagle's medium (MEM), fetal bovine serum, trypsin–EDTA, penicillin and streptomycin were purchased from GIBCO® (Invitrogen, Carlsbad, CA, USA). Cytochalasin-B (Cyt-B) and sulfanilamide were obtained from Sigma–Aldrich Co. (St. Louis, MO, USA). N-(1-Naphthyl)-ethylenediamine dihydrochloride was purchased Merck (USA). All other chemicals and reagents used were of analytical grade.
Cells and treatment
Chinese hamster lung
Cytotoxicity
Tambjamine D inhibited the proliferation of V79 cells when analyzed by the MTT assay. After 24 h of exposure, the alkaloid exhibited a strong cytotoxic effect on the V79 cell line with an IC50 of 1.2 μg/mL (CI 95% of 0.06–2.41). Based on the MTT results, tambjamine D was diluted to four different concentrations (0.6, 1.2, 2.4 and 4.8 μg/mL) corresponding to 1/2 × IC50, IC50, 2 × IC50 and 4 × IC50. The results in Fig. 2 show that tambjamine D exposure reduced significantly the number of viable cells at
Discussion
During our earlier investigation of bioactive metabolites from marine invertebrates [30], we isolated and evaluated the antimicrobial and cytotoxic activities of tambjamines A and D. While tambjamine A was essentially inactive in our assays, tambjamine D displayed antibacterial, antifungal and cytotoxic activities [13].
In this study, we investigated the biological effects of tambjamine D in cultured mammalian cells. V79 fibroblasts were used because they represent a well-characterized cell line
Acknowledgments
We wish to thank the Brazilian National Research Council (CNPq), the Claude Bernard Institute, the Research Support Foundation of Ceará (FUNCAP) and FAPESP (São Paulo) for financial support in the form of grants and fellowship awards. We are also grateful to Dr. A. Leyva for English editing of the manuscript.
References (50)
- et al.
Copper-nuclease efficiency correlates with cytotoxicity for the 4-methoxypyrrolic natural products
J. Inorg. Biochem.
(2001) - et al.
Frameshift mutations: relative roles of simple intercalation and of adduct formation
Mutat. Res.
(1981) - et al.
The influence of chemical structure on the extend and sites of carcinogens and 55 different human carcinogen exposures
Mutat. Res.
(1993) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays
J. Immunol. Methods
(1983)- et al.
Chemical and pharmacological characterization of halitoxin from Amphimedon viridis (PORIFERA) from the southeastern Brazilian coast
Comp. Biochem. Physiol.
(1996) - et al.
Malondialdehyde determination as an index of lipid peroxidation
Methods Enzymol.
(1990) - et al.
A simple technique for quantification of low levels of DNA damage in individual cells
Exp. Cell Res.
(1988) - et al.
The contribution of cytotoxicity to DNA-effects in the single cell gel test (comet assay)
Toxicol. Lett.
(1997) - et al.
The kinetics of repair of oxidative DNA damage (strand breaks and oxidized pyrimidines) in human cells
Mutat. Res.
(1995) - et al.
Evaluation of the micronucleus test using a Chinese hamster cell line as an alternative to the conventional in vitro chromosomal aberration test
Mutat. Res.
(1992)
The in vitro micronucleus technique
Mutat. Res.
The end of (cell) line: methods for the study of apoptosis in vitro
Methods Cell Biol.
Mutagenesis by chemical agents in V79 Chinese hamster cells: a review and analysis of literature
Mutat. Res.
Cytotoxicity of fungal metabolites to lepidopteran (Spodoptera frugiperda) cell line (SF-9)
J. Invertebr. Pathol.
Scavenger and antioxidant properties of prenylflavones isolated from Artocarpus heterophyllus
Free Radical Biol. Med.
Oxidative DNA modifications
Exp. Toxicol. Pathol.
Comparative study with the alkaline Comet assay and the chromosome aberration test
Mutat. Res.
The Comet assay: a comprehensive review
Mutat. Res.
Towards a validation of the micronucleus test
Mutat. Res.
In vitro genotoxicity and cell transformation assessment
Assessment of atypical DNA intercalating agents in biological and in silico systems
Mutat. Res.
Equilibrium and kinetics of rotamer interconversion in immunosuppressant prodigiosin derivatives in solution
J. Pharm. Sci.
Efficient induction of chromosome-type aberrations by topoisomerase II inhibitors closely associated with stabilization of the cleavable complex in cultured fibroblastic cells
Mutat. Res.
Gentotoxic profile of inhibitors of topoisomerase I (camptothecin) and II (etoposide) in a mitotic recombination and sex-chromosome loss somatic eye assay of Drosophila melanogaster
Mutat. Res.
Morphological study on apoptosis Hela cells induced by soyasaponis
Toxicol. In Vitro
Cited by (38)
The chemistry and chemical ecology of nudibranchs
2017, Natural Product ReportsSynthetic tambjamine analogues induce mitochondrial swelling and lysosomal dysfunction leading to autophagy blockade and necrotic cell death in lung cancer
2017, Biochemical PharmacologyCitation Excerpt :Emulating that characteristic, different anionophores have been designed and evaluated [15]. We have focused our attention on the bioactive marine alkaloids tambjamines, a class of natural anionophores that has shown interesting cytotoxic effects [16,17]. We have recently synthesized novel synthetic tambjamine analogues bearing aromatic substituents in the enamine moiety, as well as explored different substitution patterns on characteristic alkoxy group of the central pyrrole ring.
Synthesis of 2,2′-bipyrrole-5-carboxaldehydes and their application in the synthesis of B-ring functionalized prodiginines and tambjamines
2013, TetrahedronCitation Excerpt :D'Alessio et al. demonstrated that elaborating the ring-B methoxy group to provide other ethers has improved the therapeutic potential of certain PGs.5f The only other variation in ring B of PGs is recent work by Lubell's group who introduced alkylamine groups by a diversity-oriented strategy.14 Our recent work on the potent antimalarial activities of PGs,11 which to date have been limited to SAR studies of A- and C-ring functionalized PGs. With a few exceptions,5f,7,14 there have been no reports of a comprehensive series of B-ring functionalized PGs and tambjamines being prepared and evaluated for biological activities. A methodology for enhancing the diversity of 2,2′-bipyrrole-5-carboxaldehyde moiety is thus desirable to further advance SAR studies of interesting PPM products (PGs, tambjamines and etc.).
Genetic toxicology evaluation of essential oil of Alpinia zerumbet and its chemoprotective effects against H<inf>2</inf>O<inf>2</inf>-induced DNA damage in cultured human leukocytes
2012, Food and Chemical ToxicologyCitation Excerpt :According to World Health Organization (2002), about 80% of the population in developing countries rely on herbal medicines at least for their primary health care. Moreover, few plants have been scientifically assessed regarding their quality, safety and efficacy (Cavalcanti et al., 2008a). In spite of this, there have been few studies of the Brazilian medicinal flora aimed at examining potential health risks.
THE TAMBJAMINES: PYRROLYLPYRROMETHENE-CONTAINING ALKALOIDS WITH DIVERSE BIOLOGICAL PROFILES
2023, Targets in Heterocyclic Systems