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Nanomicellar Formulation of Clotrimazole Improves Its Antitumor Action toward Human Breast Cancer Cells.

Marcondes MC, Fernandes AC, Itabaiana I, de Souza RO, Sola-Penna M, Zancan P - PLoS ONE (2015)

Bottom Line: We found that nCTZ was more efficient than sCTZ at inhibiting glycolytic and other cytosolic and mitochondrial enzymes.This was especially evident on regard to antioxidant potential, which is an important cell defense against drugs that affect cell metabolism.Moreover, this water-soluble formulation of CTZ strengths its potential use as an anticancer medicine.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Oncobiologia Molecular (LabOMol), Departamento de Biotecnologia Farmacêutica (BioTecFar), Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.

ABSTRACT

Background: Although demonstrated as a selective anticancer drug, the clinical use of clotrimazole (CTZ) is limited due to its low solubility in hydrophilic fluids. Thus, we prepared a water-soluble nanomicellar formulation of CTZ (nCTZ) and tested on the human breast cancer cell line MCF-7 biology.

Methodology/principal findings: CTZ was nanoencapsulated in tween 80 micelles, which generated nanomicelles of, approximately, 17 nm of diameter. MCF-7 cells were treated with nCTZ and unencapsulated DMSO-solubilized drug (sCTZ) was used for comparison. After treatment, the cells were evaluated in terms of metabolism, proliferation, survival and structure. We found that nCTZ was more efficient than sCTZ at inhibiting glycolytic and other cytosolic and mitochondrial enzymes. Moreover, this increased activity was also observed for lactate production, intracellular ATP content, ROS production and antioxidant potential. As a consequence, nCTZ-treated MCF-7 cells displayed alterations to the plasma membrane, mitochondria and the nucleus. Finally, nCTZ induced both apoptosis and necrosis in MCF-7 cells.

Conclusions/significance: MCF-7 cells are more sensible to nCTZ than to sCTZ. This was especially evident on regard to antioxidant potential, which is an important cell defense against drugs that affect cell metabolism. Moreover, this water-soluble formulation of CTZ strengths its potential use as an anticancer medicine.

No MeSH data available.


Related in: MedlinePlus

Transmission electron microscopy of the nuclei of MCF-7 cells treated with nanomicellar CTZ.The experimental procedures are described in Materials and Methods. Panel A: non-treated control cells. Panel B: MCF-7 cells treated with 50 μM nCTZ. Panel C: MCF-7 cells treated with 100 μM nCTZ. Panel D: MCF-7 cells treated with nanomicelles prepared in the absence of CTZ. Bar = 5 μm. Images are representative of a series of at least four experiments.
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pone.0130555.g008: Transmission electron microscopy of the nuclei of MCF-7 cells treated with nanomicellar CTZ.The experimental procedures are described in Materials and Methods. Panel A: non-treated control cells. Panel B: MCF-7 cells treated with 50 μM nCTZ. Panel C: MCF-7 cells treated with 100 μM nCTZ. Panel D: MCF-7 cells treated with nanomicelles prepared in the absence of CTZ. Bar = 5 μm. Images are representative of a series of at least four experiments.

Mentions: Mitochondrial structure was also affected by nCTZ, as revealed by transmission electron microscopy (Fig 7). Control cells (Fig 7A) and the cells treated with empty nanomicelles (Fig 7D) show a normal mitochondrial profile with a double membrane and parallel cristae. Upon treatment with 50 μM nCTZ (Fig 7B), less and shorter cristae structures, a more poorly defined intermembrane space, and a less dense matrix—indicative of the loss of matrix content—were observed compared to the control. These malformations were worsened upon treatment with 100 μM nCTZ (Fig 7C); mitochondria became less elongated, cristae almost disappeared, and the matrix was even less dense. The nuclear structure was also affected by nCTZ; nuclear condensation was promoted as a function of the drug concentration (Fig 8). It can be observed that upon treatment with 50 μM nCTZ (Fig 8B), nuclear condensation is augmented compared to control cells (Fig 8A) and cells treated with empty nanomicelles (Fig 8D), and this effect increases with 100 μM nCTZ (Fig 8C).


Nanomicellar Formulation of Clotrimazole Improves Its Antitumor Action toward Human Breast Cancer Cells.

Marcondes MC, Fernandes AC, Itabaiana I, de Souza RO, Sola-Penna M, Zancan P - PLoS ONE (2015)

Transmission electron microscopy of the nuclei of MCF-7 cells treated with nanomicellar CTZ.The experimental procedures are described in Materials and Methods. Panel A: non-treated control cells. Panel B: MCF-7 cells treated with 50 μM nCTZ. Panel C: MCF-7 cells treated with 100 μM nCTZ. Panel D: MCF-7 cells treated with nanomicelles prepared in the absence of CTZ. Bar = 5 μm. Images are representative of a series of at least four experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476588&req=5

pone.0130555.g008: Transmission electron microscopy of the nuclei of MCF-7 cells treated with nanomicellar CTZ.The experimental procedures are described in Materials and Methods. Panel A: non-treated control cells. Panel B: MCF-7 cells treated with 50 μM nCTZ. Panel C: MCF-7 cells treated with 100 μM nCTZ. Panel D: MCF-7 cells treated with nanomicelles prepared in the absence of CTZ. Bar = 5 μm. Images are representative of a series of at least four experiments.
Mentions: Mitochondrial structure was also affected by nCTZ, as revealed by transmission electron microscopy (Fig 7). Control cells (Fig 7A) and the cells treated with empty nanomicelles (Fig 7D) show a normal mitochondrial profile with a double membrane and parallel cristae. Upon treatment with 50 μM nCTZ (Fig 7B), less and shorter cristae structures, a more poorly defined intermembrane space, and a less dense matrix—indicative of the loss of matrix content—were observed compared to the control. These malformations were worsened upon treatment with 100 μM nCTZ (Fig 7C); mitochondria became less elongated, cristae almost disappeared, and the matrix was even less dense. The nuclear structure was also affected by nCTZ; nuclear condensation was promoted as a function of the drug concentration (Fig 8). It can be observed that upon treatment with 50 μM nCTZ (Fig 8B), nuclear condensation is augmented compared to control cells (Fig 8A) and cells treated with empty nanomicelles (Fig 8D), and this effect increases with 100 μM nCTZ (Fig 8C).

Bottom Line: We found that nCTZ was more efficient than sCTZ at inhibiting glycolytic and other cytosolic and mitochondrial enzymes.This was especially evident on regard to antioxidant potential, which is an important cell defense against drugs that affect cell metabolism.Moreover, this water-soluble formulation of CTZ strengths its potential use as an anticancer medicine.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Oncobiologia Molecular (LabOMol), Departamento de Biotecnologia Farmacêutica (BioTecFar), Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.

ABSTRACT

Background: Although demonstrated as a selective anticancer drug, the clinical use of clotrimazole (CTZ) is limited due to its low solubility in hydrophilic fluids. Thus, we prepared a water-soluble nanomicellar formulation of CTZ (nCTZ) and tested on the human breast cancer cell line MCF-7 biology.

Methodology/principal findings: CTZ was nanoencapsulated in tween 80 micelles, which generated nanomicelles of, approximately, 17 nm of diameter. MCF-7 cells were treated with nCTZ and unencapsulated DMSO-solubilized drug (sCTZ) was used for comparison. After treatment, the cells were evaluated in terms of metabolism, proliferation, survival and structure. We found that nCTZ was more efficient than sCTZ at inhibiting glycolytic and other cytosolic and mitochondrial enzymes. Moreover, this increased activity was also observed for lactate production, intracellular ATP content, ROS production and antioxidant potential. As a consequence, nCTZ-treated MCF-7 cells displayed alterations to the plasma membrane, mitochondria and the nucleus. Finally, nCTZ induced both apoptosis and necrosis in MCF-7 cells.

Conclusions/significance: MCF-7 cells are more sensible to nCTZ than to sCTZ. This was especially evident on regard to antioxidant potential, which is an important cell defense against drugs that affect cell metabolism. Moreover, this water-soluble formulation of CTZ strengths its potential use as an anticancer medicine.

No MeSH data available.


Related in: MedlinePlus