Journal
TOXICON
Volume 119, Issue -, Pages 140-151Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.toxicon.2016.05.016
Keywords
Picocyanobacteria; Anticancer potential; Cell cycle; Apoptosis; Proteomics
Categories
Funding
- Portuguese Foundation for Science and Technology (FCT) [PTDC/MAR/102638/2008, UID/Multi/04423/2013]
- FCT [BTI/PTDC/MAR/102638/2008/2010-025]
- project MARBIOTECH within the SR&TD Integrated Program MARVALOR - Building research and innovation capacity for improved management and valorization of marine resources - Programa Operational Regional do Norte (ON.2 - O Novo Norte) [NORTE-07-0124-FEDER-000047]
- project NOVOMAR - European Regional Development Fund [0687-NOVOMAR-1-P]
- Fundação para a Ciência e a Tecnologia [PTDC/MAR/102638/2008] Funding Source: FCT
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Purpose: In this work, we analysed the potential of picoplanktonic marine cyanobacteria strains as a source of anticancer compounds by elucidating the cytotoxic mechanisms of an ethyl acetate fraction of Cyanobium sp. (LEGE06113) and the Synechocystis salina (LEGE06155) on the RKO colon adenocarcinoma cell line. Methods: Cytotoxicity was analysed by MIT. Effects on cells were evaluated by mRNA expression of cell cycle and apoptotic genes, flow cytometry (cell cycle), qualitative and quantitative fluorescence microscopy (apoptosis), and quantitative proteomics. Results: IC50 values were 27.01 and 8.03 mu g/ml for Cyanobium sp., and 37.71 and 17.17 mu g/ml for Synechocystis salina, after 24 h and 48 h, respectively. Exposure to the Cyanobium sp. fraction increased 2.5 fold BCL-2 mRNA expression (p < 0.05), and altered proteins (13, p < 0.05) belonged to apoptosis (PSMA5, PSMA7, TPT1, UBE2K), cell cycle (EIF4E, PCNA), cellular metabolism (AHSG, GLO1, ATP5H, HSP90AB1, NME1, HNRNPC) and cell structure (KRT10). Exposure to the Synechocystis salina fraction decreased 2fold CCNB1 mRNA expression (p < 0.05). Accordingly, flow cytometry demonstrated a decrease of cells in the GO/G1 and S phase (p < 0.05), indicating a cell cycle arrest at the G2/M transition. Fluorescence microscopy confirmed a higher level of apoptosis compared to the solvent control group (p < 0.01). Altered proteins (6, p < 0.05) belonged to apoptosis (HSPD1, UBE2K), protein metabolism (PKM, PDIA3) and cell structure (KRT10, KRT1). Conclusion: Since induction of cytotoxicity is a very broad parameter, the study demonstrates the potential of picocyanobacteria to produce bioactive compounds that target cancer cells via different molecular mechanisms. (C) 2016 Elsevier Ltd. All rights reserved.
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