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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

Giemsa optical microscopy of MCF-7 cells treated with nanomicellar CTZ.The experimental procedures are described in Materials and Methods. Panel A: non-treated control cells (inset: MCF-7 cells treated with nanomicelles prepared in the absence of CTZ). Panel B: MCF-7 cells treated with 50 μM nCTZ. Panel C: MCF-7 cells treated with 100 μM nCTZ. Bar = 50 μm. Images are representative of a series of at least four experiments.
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pone.0130555.g005: Giemsa optical microscopy of MCF-7 cells treated with nanomicellar CTZ.The experimental procedures are described in Materials and Methods. Panel A: non-treated control cells (inset: MCF-7 cells treated with nanomicelles prepared in the absence of CTZ). Panel B: MCF-7 cells treated with 50 μM nCTZ. Panel C: MCF-7 cells treated with 100 μM nCTZ. Bar = 50 μm. Images are representative of a series of at least four experiments.

Mentions: nCTZ is devastating to MCF-7 cell structure, as revealed by distinct microscopy techniques. Giemsa staining of MCF-7 cells treated for 24 hours with nCTZ revealed profoundly affected cell morphology (Fig 5). Compared to control cells (Fig 5, panel A), the treatment of MCF-7 cells with 50 μM nCTZ (Fig 5, panel B) converted the stellar-shaped MCF-7 cells into an elongated fusiform morphology lacking protrusions. Moreover, after treatment with 100 μM nCTZ, MFC-7 cells become more spherical/elliptical, resembling primitive undifferentiated cells (Fig 5, panel C). The inset to the major panel A of Fig 5 shows MCF-7 cells treated for 24 hours with nanomicelles prepared in the absence of CTZ, and it is clear that they have no effect on cell morphology. These results are corroborated by scanning electron microscopy, which reveals the above alterations in more detail (Fig 6). This technique reveals that the plasma membrane of untreated control cells (Fig 6A) is homogeneously rough. This pattern is altered by treatment with 50 μM nCTZ (Fig 6B); the roughness becomes more irregular, and the plasma membrane presents bubbles on its surface (also observed as dense regions in Fig 5). These results are enhanced by treatment with 100 μM nCTZ (Fig 6C), which, in addition to the cell shape deformation, clearly disrupts the plasma membrane. Detail in panel C reveals that the plasma membrane integrity is damaged, with membrane fragments around the cells. The lack of effect with empty nanomicelles was also demonstrated by scanning electron microscopy in the inset of Fig 6A.


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)

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

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

pone.0130555.g005: Giemsa optical microscopy of MCF-7 cells treated with nanomicellar CTZ.The experimental procedures are described in Materials and Methods. Panel A: non-treated control cells (inset: MCF-7 cells treated with nanomicelles prepared in the absence of CTZ). Panel B: MCF-7 cells treated with 50 μM nCTZ. Panel C: MCF-7 cells treated with 100 μM nCTZ. Bar = 50 μm. Images are representative of a series of at least four experiments.
Mentions: nCTZ is devastating to MCF-7 cell structure, as revealed by distinct microscopy techniques. Giemsa staining of MCF-7 cells treated for 24 hours with nCTZ revealed profoundly affected cell morphology (Fig 5). Compared to control cells (Fig 5, panel A), the treatment of MCF-7 cells with 50 μM nCTZ (Fig 5, panel B) converted the stellar-shaped MCF-7 cells into an elongated fusiform morphology lacking protrusions. Moreover, after treatment with 100 μM nCTZ, MFC-7 cells become more spherical/elliptical, resembling primitive undifferentiated cells (Fig 5, panel C). The inset to the major panel A of Fig 5 shows MCF-7 cells treated for 24 hours with nanomicelles prepared in the absence of CTZ, and it is clear that they have no effect on cell morphology. These results are corroborated by scanning electron microscopy, which reveals the above alterations in more detail (Fig 6). This technique reveals that the plasma membrane of untreated control cells (Fig 6A) is homogeneously rough. This pattern is altered by treatment with 50 μM nCTZ (Fig 6B); the roughness becomes more irregular, and the plasma membrane presents bubbles on its surface (also observed as dense regions in Fig 5). These results are enhanced by treatment with 100 μM nCTZ (Fig 6C), which, in addition to the cell shape deformation, clearly disrupts the plasma membrane. Detail in panel C reveals that the plasma membrane integrity is damaged, with membrane fragments around the cells. The lack of effect with empty nanomicelles was also demonstrated by scanning electron microscopy in the inset of Fig 6A.

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