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Comparative assessment of the apoptotic potential of silver nanoparticles synthesized by Bacillus tequilensis and Calocybe indica in MDA-MB-231 human breast cancer cells: targeting p53 for anticancer therapy.

Gurunathan S, Park JH, Han JW, Kim JH - Int J Nanomedicine (2015)

Bottom Line: This is especially true in the area of nanomedicine, due to physicochemical properties, such as mechanical, chemical, magnetic, optical, and electrical properties, compared with bulk materials.The first goal of this study was to produce silver nanoparticles (AgNPs) using two different biological resources as reducing agents, Bacillus tequilensis and Calocybe indica.Cells pretreated with pifithrin-alpha were protected from p53-mediated AgNPs-induced toxicity.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, Republic of Korea.

ABSTRACT

Background: Recently, the use of nanotechnology has been expanding very rapidly in diverse areas of research, such as consumer products, energy, materials, and medicine. This is especially true in the area of nanomedicine, due to physicochemical properties, such as mechanical, chemical, magnetic, optical, and electrical properties, compared with bulk materials. The first goal of this study was to produce silver nanoparticles (AgNPs) using two different biological resources as reducing agents, Bacillus tequilensis and Calocybe indica. The second goal was to investigate the apoptotic potential of the as-prepared AgNPs in breast cancer cells. The final goal was to investigate the role of p53 in the cellular response elicited by AgNPs.

Methods: The synthesis and characterization of AgNPs were assessed by various analytical techniques, including ultraviolet-visible (UV-vis) spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS), and transmission electron microscopy (TEM). The apoptotic efficiency of AgNPs was confirmed using a series of assays, including cell viability, leakage of lactate dehydrogenase (LDH), production of reactive oxygen species (ROS), DNA fragmentation, mitochondrial membrane potential, and Western blot.

Results: The absorption spectrum of the yellow AgNPs showed the presence of nanoparticles. XRD and FTIR spectroscopy results confirmed the crystal structure and biomolecules involved in the synthesis of AgNPs. The AgNPs derived from bacteria and fungi showed distinguishable shapes, with an average size of 20 nm. Cell viability assays suggested a dose-dependent toxic effect of AgNPs, which was confirmed by leakage of LDH, activation of ROS, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells in MDA-MB-231 breast cancer cells. Western blot analyses revealed that AgNPs induce cellular apoptosis via activation of p53, p-Erk1/2, and caspase-3 signaling, and downregulation of Bcl-2. Cells pretreated with pifithrin-alpha were protected from p53-mediated AgNPs-induced toxicity.

Conclusion: We have demonstrated a simple approach for the synthesis of AgNPs using the novel strains B. tequilensis and C. indica, as well as their mechanism of cell death in a p53-dependent manner in MDA-MB-231 human breast cancer cells. The present findings could provide insight for the future development of a suitable anticancer drug, which may lead to the development of novel nanotherapeutic molecules for the treatment of cancers.

No MeSH data available.


Related in: MedlinePlus

Synthesis and characterization of B-AgNPs and F-AgNPs using culture supernatant of Bacillus tequilensis and culture filtrate of milky mushroom, respectively.Notes: The formation of AgNPs was confirmed using UV-vis spectroscopy. The absorption spectra of B-AgNPs and F-AgNPs exhibited a strong broad peak at 410 nm, and observation of such a band is assigned to surface plasmon resonance of the particles.Abbreviations: AgNPs, silver nanoparticles; B-AgNPs, bacterium-derived AgNPs; F-AgNPs, fungus-derived AgNPs; UV-vis, ultraviolet-visible.
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f1-ijn-10-4203: Synthesis and characterization of B-AgNPs and F-AgNPs using culture supernatant of Bacillus tequilensis and culture filtrate of milky mushroom, respectively.Notes: The formation of AgNPs was confirmed using UV-vis spectroscopy. The absorption spectra of B-AgNPs and F-AgNPs exhibited a strong broad peak at 410 nm, and observation of such a band is assigned to surface plasmon resonance of the particles.Abbreviations: AgNPs, silver nanoparticles; B-AgNPs, bacterium-derived AgNPs; F-AgNPs, fungus-derived AgNPs; UV-vis, ultraviolet-visible.

Mentions: The synthesis of AgNPs was monitored by UV-vis spectroscopy, which is a reliable and valuable technique for verifying the formation of metal nanoparticles, provided that surface plasmon resonance exists for the metal.33 It is known from the spectra that the silver surface plasmon resonance band occurs at 420 nm.36–38Figure 1 shows the UV-vis spectrum of synthesized AgNPs from 5 mM AgNO3 with culture supernatant of bacteria or extracellular filtrate of the milky mushroom. It is observed that the band corresponding to surface plasmon resonance occurs at 410 nm, which clearly indicates the formation of AgNPs in the reaction mixture.33,36–40 Similarly, Anthony et al reported AgNPs derived from culture supernatant of Bacillus mariflavi.7 Mukherjee et al demonstrated synthesis of AgNPs using culture filtrate of Verticillium.3 Extracellular synthesis of AgNPs has been carried out using the aqueous extract of edible oyster mushroom and Ganoderma neo-japonicum as a reducing agent.10,41 A strong and broad surface plasmon peak located at 410 nm was observed for the AgNPs prepared using both culture supernatant of bacteria and mycelia extract of milky mushroom.10,36 The strong surface plasmon resonance centered at 410 nm clearly indicates the formation of AgNPs, which were extremely stable, with no evidence of flocculation of the particles even after 3 months.36 The band approximately 410 nm suggests that the particles were well-dispersed, without aggregation.10 The long-term stability of the AgNPs solution may be due to the proteins in the culture supernatant of bacteria or mushroom extract acting as capping agents.


Comparative assessment of the apoptotic potential of silver nanoparticles synthesized by Bacillus tequilensis and Calocybe indica in MDA-MB-231 human breast cancer cells: targeting p53 for anticancer therapy.

Gurunathan S, Park JH, Han JW, Kim JH - Int J Nanomedicine (2015)

Synthesis and characterization of B-AgNPs and F-AgNPs using culture supernatant of Bacillus tequilensis and culture filtrate of milky mushroom, respectively.Notes: The formation of AgNPs was confirmed using UV-vis spectroscopy. The absorption spectra of B-AgNPs and F-AgNPs exhibited a strong broad peak at 410 nm, and observation of such a band is assigned to surface plasmon resonance of the particles.Abbreviations: AgNPs, silver nanoparticles; B-AgNPs, bacterium-derived AgNPs; F-AgNPs, fungus-derived AgNPs; UV-vis, ultraviolet-visible.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494182&req=5

f1-ijn-10-4203: Synthesis and characterization of B-AgNPs and F-AgNPs using culture supernatant of Bacillus tequilensis and culture filtrate of milky mushroom, respectively.Notes: The formation of AgNPs was confirmed using UV-vis spectroscopy. The absorption spectra of B-AgNPs and F-AgNPs exhibited a strong broad peak at 410 nm, and observation of such a band is assigned to surface plasmon resonance of the particles.Abbreviations: AgNPs, silver nanoparticles; B-AgNPs, bacterium-derived AgNPs; F-AgNPs, fungus-derived AgNPs; UV-vis, ultraviolet-visible.
Mentions: The synthesis of AgNPs was monitored by UV-vis spectroscopy, which is a reliable and valuable technique for verifying the formation of metal nanoparticles, provided that surface plasmon resonance exists for the metal.33 It is known from the spectra that the silver surface plasmon resonance band occurs at 420 nm.36–38Figure 1 shows the UV-vis spectrum of synthesized AgNPs from 5 mM AgNO3 with culture supernatant of bacteria or extracellular filtrate of the milky mushroom. It is observed that the band corresponding to surface plasmon resonance occurs at 410 nm, which clearly indicates the formation of AgNPs in the reaction mixture.33,36–40 Similarly, Anthony et al reported AgNPs derived from culture supernatant of Bacillus mariflavi.7 Mukherjee et al demonstrated synthesis of AgNPs using culture filtrate of Verticillium.3 Extracellular synthesis of AgNPs has been carried out using the aqueous extract of edible oyster mushroom and Ganoderma neo-japonicum as a reducing agent.10,41 A strong and broad surface plasmon peak located at 410 nm was observed for the AgNPs prepared using both culture supernatant of bacteria and mycelia extract of milky mushroom.10,36 The strong surface plasmon resonance centered at 410 nm clearly indicates the formation of AgNPs, which were extremely stable, with no evidence of flocculation of the particles even after 3 months.36 The band approximately 410 nm suggests that the particles were well-dispersed, without aggregation.10 The long-term stability of the AgNPs solution may be due to the proteins in the culture supernatant of bacteria or mushroom extract acting as capping agents.

Bottom Line: This is especially true in the area of nanomedicine, due to physicochemical properties, such as mechanical, chemical, magnetic, optical, and electrical properties, compared with bulk materials.The first goal of this study was to produce silver nanoparticles (AgNPs) using two different biological resources as reducing agents, Bacillus tequilensis and Calocybe indica.Cells pretreated with pifithrin-alpha were protected from p53-mediated AgNPs-induced toxicity.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Biotechnology, Konkuk University, Seoul, Republic of Korea.

ABSTRACT

Background: Recently, the use of nanotechnology has been expanding very rapidly in diverse areas of research, such as consumer products, energy, materials, and medicine. This is especially true in the area of nanomedicine, due to physicochemical properties, such as mechanical, chemical, magnetic, optical, and electrical properties, compared with bulk materials. The first goal of this study was to produce silver nanoparticles (AgNPs) using two different biological resources as reducing agents, Bacillus tequilensis and Calocybe indica. The second goal was to investigate the apoptotic potential of the as-prepared AgNPs in breast cancer cells. The final goal was to investigate the role of p53 in the cellular response elicited by AgNPs.

Methods: The synthesis and characterization of AgNPs were assessed by various analytical techniques, including ultraviolet-visible (UV-vis) spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS), and transmission electron microscopy (TEM). The apoptotic efficiency of AgNPs was confirmed using a series of assays, including cell viability, leakage of lactate dehydrogenase (LDH), production of reactive oxygen species (ROS), DNA fragmentation, mitochondrial membrane potential, and Western blot.

Results: The absorption spectrum of the yellow AgNPs showed the presence of nanoparticles. XRD and FTIR spectroscopy results confirmed the crystal structure and biomolecules involved in the synthesis of AgNPs. The AgNPs derived from bacteria and fungi showed distinguishable shapes, with an average size of 20 nm. Cell viability assays suggested a dose-dependent toxic effect of AgNPs, which was confirmed by leakage of LDH, activation of ROS, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells in MDA-MB-231 breast cancer cells. Western blot analyses revealed that AgNPs induce cellular apoptosis via activation of p53, p-Erk1/2, and caspase-3 signaling, and downregulation of Bcl-2. Cells pretreated with pifithrin-alpha were protected from p53-mediated AgNPs-induced toxicity.

Conclusion: We have demonstrated a simple approach for the synthesis of AgNPs using the novel strains B. tequilensis and C. indica, as well as their mechanism of cell death in a p53-dependent manner in MDA-MB-231 human breast cancer cells. The present findings could provide insight for the future development of a suitable anticancer drug, which may lead to the development of novel nanotherapeutic molecules for the treatment of cancers.

No MeSH data available.


Related in: MedlinePlus