Methods: Several compounds were synthesized and their molecular identity was confirmed using nuclear magnetic resonance. Potential anticancer properties were determined using cytopathogenicity assays and growth inhibition assays using cervical cancer cells (HeLa). Cells were incubated with different concentrations of compounds belonging to Benzodioxane, Naphthalene diimide, Aminophenol derivatives and Porphyrins and effects were determined. HeLa cells cytopathogenicity was determined by measuring lactate dehydrogenase release using cytotoxicity detection assay. Growth inhibition assays were performed by incubating 50% semi-confluent HeLa cells with Benzodioxane, Naphthalene diimide, Aminophenol derivatives and Porphyrin compounds and HeLa cell proliferation was observed. Growth inhibition and host cell death were compared in the presence and absence of drugs.
Results: Cytopathogenicity assays showed that the selected compounds were cytotoxic against HeLa cells, killing up to 90% of cells. Growth inhibition assays exhibited 100% growth inhibition. These effects are likely via oxidative stress, production of reactive oxygen species, changes in cytosolic and intracellular calcium/adenine nucleotide homeostasis, inhibition of ribonucleotide reductase/cyclooxygenase and/or glutathione depletion.
Conclusions: Benzodioxane, Naphthalene diimide, Aminophenol derivatives and Porphyrins exhibited potent anticancer properties. These findings are promising and should pave the way in the rationale development of anticancer drugs. Using different cancer cell lines, future studies will determine their potential as anti-tumour agents as well as their precise molecular mode of action.
METHODS: In this study, various species of vertebrates and invertebrates were procured including Columba livia, Gallus gallus domesticus, Varanus salvator, Cuora kamamora amboinensis, Reticulatus malayanus, Oreochromis mossambicus, Rattus rattus, American bullfrog, Donax sp., Polymesoda coaxans, Tenebrio molitor, Lumbricus terrestris, Blatta lateralis, Grammostola rosea, and Penaeus monodon. Species were dissected and their organ lysates/sera/haemolymph were prepared. Cytotoxicity assays were performed using Prostate Cancer cells (PC3), Henrietta Lacks cervical adenocarcinoma cells (HeLa) and human breast adenocarcinoma cells (MCF7) as well as human keratinized skin cells (Hacat), by measuring lactate dehydrogenase release as an indicator for cell death. Growth inhibition assays were performed to determine the effects on cancer cell proliferation. Liquid Chromatography-Mass Spectrometry (LC-MS/MS) was performed for molecular identification.
RESULTS: The results revealed that body lysates of Polymesoda coaxans demonstrated more than 99% growth inhibition of all cancer cell lines tested but not on normal Hacat cells. More importantly, the serum of M. reticulatus abolished growth and produced cytotoxicity. Hence these samples were subjected to Liquid Chromatography- Mass Spectrometry (LC-MS/MS), which detected 81 small molecules and putatively identified 20 molecules when matched against the METLIN database. Out of 1094 peptides, 21 peptides were identified, while 1074 peptides were categorized as novel peptides. Based on properties such as peptide amino acid composition, binary profile, dipeptide composition and pseudo-amino acid composition, 306 potential peptides were identified.
CONCLUSION: To our knowledge, here for the first time, we report a comprehensive analysis of sera exhibiting cytotoxicity against cancer cell lines tested and identified several molecules using LC-MS/MS.
METHODS: Using purified compounds, assays were performed to determine their effects against cancer cell lines using growth inhibition assays, cytotoxicity assays, and cell survival assays against HeLa, PC3 and MCF7 cells.
RESULTS: The results showed that the selected small molecules L-Methionine, Rofecoxib, and Artocarpin suppressed the growth of more than 90% PC3 cells at 40µM. Similarly, Valdecoxib alone and in combination with other molecules exhibited potent growth inhibition and cytotoxicity against cancer cells tested. Peptide from the serum of M. reticulatus, demonstrated selective cytotoxicity against cancer cells without inhibiting the growth of normal cells.
CONCLUSION: These findings are significant and provide a basis for the rational development of therapeutic anticancer agents, however intensive research is needed to determine in vivo effects of the identified molecules together with their mode of action to realize these expectations.
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METHODS: Faecal and gut microbiota of Columbia livia were isolated, identified and conditioned media were prepared containing metabolites. Growth inhibition, lactate dehydrogenase cytotoxicity and cell survival assays were accomplished against cervical cancer cells. Next, liquid-chromatography mass spectrometry was conducted to elucidate the molecules present.
RESULTS: A plethora of bacteria from faecal matter and gastrointestinal tract were isolated. Selected conditioned media exhibited potent anticancer effects and displayed cytotoxicity to cervical cancer cells at IC50 concentration of 10.65 and 15.19 µg/ml. Moreover, cells treated with conditioned media exhibited morphological changes, including cell shrinking and rounding; indicative of apoptosis, when compared to untreated cells. A total of 111 and 71 molecules were revealed from these gut and faecal metabolites. The identity of 60 molecules were revealed including, dihydroxymelphalan. Nonetheless, 122 molecules remain unidentified and are the subject of future studies.
CONCLUSION: These findings suggest that gut bacteria of Columbia livia possess molecules, which may have anticancer activities. Further in silico testing and/or high throughput screening will determine potential anticancer properties of these molecules.
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Methods: Crocodylus porosus was procured, blood collected, dissected and lysates prepared from internal organs. Organ lysates and sera were tested for growth inhibition, cytotoxic effects and cell survival against HeLa, PC3 and MCF7 cells and subjected to liquid chromatography mass spectrometry. RNA transcriptome analysis and differential gene analysis were performed using Galaxy Bioinformatics.
Results: Sera exhibited potent growth inhibition and cytotoxic effects against cancer cells. 80 molecules were detected from C. porosus and 19 molecules were putatively identified. Additionally, more than 100 potential anticancer peptides were identified from sera using bioinformatics based on peptide amino acid composition, binary profile, dipeptide composition and pseudo-amino acid composition. Following transcriptome analysis, 14 genes in treated HeLa cells, 51 genes in treated MCF7 cells and 2 genes in treated PC3 cells, were found to be expressed, compared with untreated controls.
Conclusion: Animals residing in polluted milieus are an unexploited source for prospective pharmaceutical drugs, and could lead to identification of novel antitumour compound(s) and/or further understanding of the mechanisms of cancer resistance.
MATERIALS AND METHODS: A PubMed search using antibacterials, antimicrobials, invertebrates, and natural products as keywords was carried out. In addition, we consulted conference proceedings, original unpublished research undertaken in our laboratories, and discussions in specific forums.
RESULTS: Representative of a stupefying 95% of the fauna, invertebrates are fascinating organisms which have evolved strategies to survive germ-infested environments, yet they have largely been ignored. Since invertebrates such as cockroaches inhabit hazardous environments which are rampant with pathogens, they must have developed defense mechanisms to circumvent infections. This is corroborated by the presence of antimicrobial molecules in the nervous systems and hemolymph of cockroaches. Antimicrobial compounds have also been unraveled from the nervous, adipose, and salivary glandular tissues of locusts. Interestingly, the venoms of arthropods including ants, scorpions, and spiders harbor toxins, but also possess multiple antimicrobials.
CONCLUSION: These findings have rekindled the hopes for newer and enhanced therapeutic agents derived from a plentiful and diverse resource to combat fatal infectious diseases. Such antimicrobials from unusual sources can potentially be translated into clinical practice, however intensive research is needed over the next several years to realize these expectations.