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Seaweed fucoidan-stabilized gold nanoparticles and their antitumor activities
Abstracts of Pharma Nutrition 2013 / PharmaNutrition 2 (2014) 75–119
functions. Using the human epithelial intestinal cell line Caco-2 in a transwell system as an in vitro model, we studied the effect of DON in the presence and absence of galacto-oligosaccharides (scGOS). Parameters assessed included transepithelial electrical resistance (TEER), paracellular tracer flux, expression of tight junction proteins, cytokine release and overall cell viability. Interestingly, when a DON challenged non-permeable Caco-2 monolayer was preincubated with scGOS for 24 h, TEER and paracellular transport of the marker lucifer yellow remained unaffected. In parallel, the levels of claudin-3 mRNA expression were not affected by DON exposure in the presence of scGOS. Furthermore, the increase in epithelial IL-8 mRNA expression and in IL-8 secretion, observed after DON exposure, was counteracted by scGOS. In conclusion, we demonstrated with this in vitro model that scGOS protects and restores the epithelial barrier dysfunction exerted by the food contaminant DON. In addition, the hypothesis that scGOS has antiinflammatory properties was confirmed by demonstrating that the DON-induced production of IL-8 was counteracted by scGOS. Taken together, these findings indicate that dietary oligosaccharides may be a promising approach to mitigate the effects of undesirable dietary mycotoxin exposure. Keywords: Deoxynivalenol; Oligosaccharides; Intestinal barrier; IL-8 http://dx.doi.org/10.1016/j.phanu.2013.11.018 [O5] From phyto-chemoprevention to discovery of novel markers and targets for therapy of breast cancer A. Ouhtit 1 , S. Shanmuganathan 1,∗ , R. Gaur 1 , M.E. Abdraboh 1 , A. Hollenbach 2 , I. Gupta 1 , M.H.G. Raj 2 1 2
Sultan Qaboos University, Oman Louisiana State University Health Science Center, USA
Background: Conventional Medicine faces serious challenges in cancer treatment such as drug resistance and lack of specificity for the target. Hence, Complementary Alternative Medicine is increasingly being practiced worldwide due to its safety and beneficial therapeutic effects. Our preliminary studies revealed that a combination of Indole-3-Carbinol and Resveratrol synergized, at bioavailable doses synergized and induced apoptosis of ovarian cancer cell lines. Therefore, we hypothesized that a super combination (SC) of additional known phytochemicals used at bioavailable levels could synergize to induce 100% clearance of breast cancer (BC) cells in vitro and microarray analysis would identify potential genes for targeted therapy of BC. Methods: Mesenchymal Stems cells (MSC control) and various BC cell lines were treated with a combination of various well established pro-apoptotic phytochemicals at bioavailable levels. Alamar Blue and Flow Cytometry assays were used to determine cell proliferation and cell death. Wound healing and invasion assays were used to examine cell migration/invasion through Matrigel. Western Blot analysis was used to examine the expression of genes associated with cell cycle, apoptosis and cell motility. Results: More than 20 phytochemicals were tested in various combinations, and a SC of six compounds synergized and induced 100% BC cell death as compared to control MSCs. Furthermore, the SC down-regulated the expression of specific genes in the BC cell lines, including PCNA, Rb, CDK4, Bcl-2, SVV, and CD44. More interestingly, microarray analysis revealed several differentially expressed key genes underpinning SC-promoted BC cell death and motility. Conclusion: The present study identified a SC of phytochemicals that induced 100% BC cell death and determined key genes
81
that underpin SC-mediated cell death and cell motility. Our ongoing experiments aim to validate this data in in vivo mouse models. Keywords: Complementary alternative medicine; Phytochemicals; Combination; Microarray http://dx.doi.org/10.1016/j.phanu.2013.11.019 [O6] Seaweed fucoidan-stabilized gold nanoparticles and their antitumor activities S. Soisuwan 1 , K. Lirdprapamongko 1,2 , W. Warisnoicharoen 1,∗ , J. Svasti 2 1 2
Chulalongkorn University, Thailand Chulabhorn Research Institute, Thailand
Fucoidans are sulphated polysaccharides extracted from brown seaweed. They have been recently employed as a nutritional supplement due to their pharmacological activities including antiinflammation, anti-peptic ulcer, anti-viral and anti-tumor activities. Truly, nano-sized particles preferentially accumulate at tumor sites leading to enhanced permeability and retention (EPR) effects, hence usage of fucoidan and nanoparticles in combination might provide a promising way for cancer treatment. According to biocompatibility, stability and size-controllability of gold nanoparticles, they were subjected to be synthesized using fucoidans as a stabilizer. The study was aimed to investigate the antitumor activities of fucoidan-stabilized gold nanoparticles (GNP) in human lung adenocarcinoma (A549) cells. GNP were synthesized using fucoidan extract from brown seaweed Cladosiphon okamuranus at a weight ratio of 1:20 gold atom to fucoidans and characterized for their size distribution and morphology by transmission electron microscopy. The cells exposed to various concentrations of GNP were determined for viability and anoikis resistance using a MTT assay. The effect of GNP on cell adhesion, cell invasion and migration was observed using the medium containing human embryonic lung fibroblast (MRC-5) cells as a chemoattractant. The results showed that GNP were spherical mono-dispersed particle with average sizes of 8.54 ± 2.96 nm. GNP could decrease the A549 cell survival more than the fucoidans and no anoikis resistance was observed. GNP had a slight inhibitory effect (∼16% inhibition) on A549 cell invasion; this effect might come from their anti-adhesive property to fibronectin (∼30% inhibition). Moreover, the level of matrix metalloproteinase-2 enzyme in the cells treated with GNP was repressed (∼60% inhibition). Conclusively, fucoidans from C. okamuranus were able to stabilize gold nanoparticles. The synthesized nanoparticles, GNP, could reduce the cell viability as well as inhibit cell adhesion and invasion. Accordingly, GNP might be a challenged compound for cancer therapy as well as the prevention of cancer metastasis. Keywords: Fucoidan; Gold nanoparticles; Anti-tumor; A549 cells http://dx.doi.org/10.1016/j.phanu.2013.11.020
functions. Using the human epithelial intestinal cell line Caco-2 in a transwell system as an in vitro model, we studied the effect of DON in the presence and absence of galacto-oligosaccharides (scGOS). Parameters assessed included transepithelial electrical resistance (TEER), paracellular tracer flux, expression of tight junction proteins, cytokine release and overall cell viability. Interestingly, when a DON challenged non-permeable Caco-2 monolayer was preincubated with scGOS for 24 h, TEER and paracellular transport of the marker lucifer yellow remained unaffected. In parallel, the levels of claudin-3 mRNA expression were not affected by DON exposure in the presence of scGOS. Furthermore, the increase in epithelial IL-8 mRNA expression and in IL-8 secretion, observed after DON exposure, was counteracted by scGOS. In conclusion, we demonstrated with this in vitro model that scGOS protects and restores the epithelial barrier dysfunction exerted by the food contaminant DON. In addition, the hypothesis that scGOS has antiinflammatory properties was confirmed by demonstrating that the DON-induced production of IL-8 was counteracted by scGOS. Taken together, these findings indicate that dietary oligosaccharides may be a promising approach to mitigate the effects of undesirable dietary mycotoxin exposure. Keywords: Deoxynivalenol; Oligosaccharides; Intestinal barrier; IL-8 http://dx.doi.org/10.1016/j.phanu.2013.11.018 [O5] From phyto-chemoprevention to discovery of novel markers and targets for therapy of breast cancer A. Ouhtit 1 , S. Shanmuganathan 1,∗ , R. Gaur 1 , M.E. Abdraboh 1 , A. Hollenbach 2 , I. Gupta 1 , M.H.G. Raj 2 1 2
Sultan Qaboos University, Oman Louisiana State University Health Science Center, USA
Background: Conventional Medicine faces serious challenges in cancer treatment such as drug resistance and lack of specificity for the target. Hence, Complementary Alternative Medicine is increasingly being practiced worldwide due to its safety and beneficial therapeutic effects. Our preliminary studies revealed that a combination of Indole-3-Carbinol and Resveratrol synergized, at bioavailable doses synergized and induced apoptosis of ovarian cancer cell lines. Therefore, we hypothesized that a super combination (SC) of additional known phytochemicals used at bioavailable levels could synergize to induce 100% clearance of breast cancer (BC) cells in vitro and microarray analysis would identify potential genes for targeted therapy of BC. Methods: Mesenchymal Stems cells (MSC control) and various BC cell lines were treated with a combination of various well established pro-apoptotic phytochemicals at bioavailable levels. Alamar Blue and Flow Cytometry assays were used to determine cell proliferation and cell death. Wound healing and invasion assays were used to examine cell migration/invasion through Matrigel. Western Blot analysis was used to examine the expression of genes associated with cell cycle, apoptosis and cell motility. Results: More than 20 phytochemicals were tested in various combinations, and a SC of six compounds synergized and induced 100% BC cell death as compared to control MSCs. Furthermore, the SC down-regulated the expression of specific genes in the BC cell lines, including PCNA, Rb, CDK4, Bcl-2, SVV, and CD44. More interestingly, microarray analysis revealed several differentially expressed key genes underpinning SC-promoted BC cell death and motility. Conclusion: The present study identified a SC of phytochemicals that induced 100% BC cell death and determined key genes
81
that underpin SC-mediated cell death and cell motility. Our ongoing experiments aim to validate this data in in vivo mouse models. Keywords: Complementary alternative medicine; Phytochemicals; Combination; Microarray http://dx.doi.org/10.1016/j.phanu.2013.11.019 [O6] Seaweed fucoidan-stabilized gold nanoparticles and their antitumor activities S. Soisuwan 1 , K. Lirdprapamongko 1,2 , W. Warisnoicharoen 1,∗ , J. Svasti 2 1 2
Chulalongkorn University, Thailand Chulabhorn Research Institute, Thailand
Fucoidans are sulphated polysaccharides extracted from brown seaweed. They have been recently employed as a nutritional supplement due to their pharmacological activities including antiinflammation, anti-peptic ulcer, anti-viral and anti-tumor activities. Truly, nano-sized particles preferentially accumulate at tumor sites leading to enhanced permeability and retention (EPR) effects, hence usage of fucoidan and nanoparticles in combination might provide a promising way for cancer treatment. According to biocompatibility, stability and size-controllability of gold nanoparticles, they were subjected to be synthesized using fucoidans as a stabilizer. The study was aimed to investigate the antitumor activities of fucoidan-stabilized gold nanoparticles (GNP) in human lung adenocarcinoma (A549) cells. GNP were synthesized using fucoidan extract from brown seaweed Cladosiphon okamuranus at a weight ratio of 1:20 gold atom to fucoidans and characterized for their size distribution and morphology by transmission electron microscopy. The cells exposed to various concentrations of GNP were determined for viability and anoikis resistance using a MTT assay. The effect of GNP on cell adhesion, cell invasion and migration was observed using the medium containing human embryonic lung fibroblast (MRC-5) cells as a chemoattractant. The results showed that GNP were spherical mono-dispersed particle with average sizes of 8.54 ± 2.96 nm. GNP could decrease the A549 cell survival more than the fucoidans and no anoikis resistance was observed. GNP had a slight inhibitory effect (∼16% inhibition) on A549 cell invasion; this effect might come from their anti-adhesive property to fibronectin (∼30% inhibition). Moreover, the level of matrix metalloproteinase-2 enzyme in the cells treated with GNP was repressed (∼60% inhibition). Conclusively, fucoidans from C. okamuranus were able to stabilize gold nanoparticles. The synthesized nanoparticles, GNP, could reduce the cell viability as well as inhibit cell adhesion and invasion. Accordingly, GNP might be a challenged compound for cancer therapy as well as the prevention of cancer metastasis. Keywords: Fucoidan; Gold nanoparticles; Anti-tumor; A549 cells http://dx.doi.org/10.1016/j.phanu.2013.11.020