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In vitro cytotoxicity analysis of doxorubicin-loaded/superparamagnetic iron oxide colloidal nanoassemblies on MCF7 and NIH3T3 cell lines.

Tomankova K, Polakova K, Pizova K, Binder S, Havrdova M, Kolarova M, Kriegova E, Zapletalova J, Malina L, Horakova J, Malohlava J, Kolokithas-Ntoukas A, Bakandritsos A, Kolarova H, Zboril R - Int J Nanomedicine (2015)

Bottom Line: For proper analysis and understanding of cell behavior after administration of MagAlg-DOX compared with free DOX, a complex set of in vitro tests, including production of reactive oxygen species, comet assay, cell cycle determination, gene expression, and cellular uptake, were utilized.It was found that the cytotoxic effect of MagAlg-DOX system is delayed compared to free DOX in both cell lines.We discovered that nanoparticles can attenuate or even inhibit the effect of DOX, particularly in the tumor MCF7 cell line.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biophysics, Institute of Translation Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.

ABSTRACT
One of the promising strategies for improvement of cancer treatment is based on magnetic drug delivery systems, thus avoiding side effects of standard chemotherapies. Superparamagnetic iron oxide (SPIO) nanoparticles have ideal properties to become a targeted magnetic drug delivery contrast probes, named theranostics. We worked with SPIO condensed colloidal nanocrystal clusters (MagAlg) prepared through a new soft biomineralization route in the presence of alginate as the polymeric shell and loaded with doxorubicin (DOX). The aim of this work was to study the in vitro cytotoxicity of these new MagAlg-DOX systems on mouse fibroblast and breast carcinoma cell lines. For proper analysis and understanding of cell behavior after administration of MagAlg-DOX compared with free DOX, a complex set of in vitro tests, including production of reactive oxygen species, comet assay, cell cycle determination, gene expression, and cellular uptake, were utilized. It was found that the cytotoxic effect of MagAlg-DOX system is delayed compared to free DOX in both cell lines. This was attributed to the different mechanism of internalization of DOX and MagAlg-DOX into the cells, together with the fact that the drug is strongly bound on the drug nanocarriers. We discovered that nanoparticles can attenuate or even inhibit the effect of DOX, particularly in the tumor MCF7 cell line. This is a first comprehensive study on the cytotoxic effect of DOX-loaded SPIO compared with free DOX on healthy and cancer cell lines, as well as on the induced changes in gene expression.

No MeSH data available.


Related in: MedlinePlus

Fold change of C-MYC (A and B) and C-FOS (C and D) expression on MCF7 cells (left graphs) and NIH3T3 cells (right graphs) in the course of 0 μM, 0.5 μM, and 5 μM in DOX concentration and with effect of MagAlg–DOX nanocarrier.Notes: Data represent mean and standard error from three independent measurements. Positive (*) and negative (•) significance were determined using ANOVA and Dunnet post hoc test.Abbreviation: DOX, doxorubicin.
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f7-ijn-10-949: Fold change of C-MYC (A and B) and C-FOS (C and D) expression on MCF7 cells (left graphs) and NIH3T3 cells (right graphs) in the course of 0 μM, 0.5 μM, and 5 μM in DOX concentration and with effect of MagAlg–DOX nanocarrier.Notes: Data represent mean and standard error from three independent measurements. Positive (*) and negative (•) significance were determined using ANOVA and Dunnet post hoc test.Abbreviation: DOX, doxorubicin.

Mentions: We observed downregulation of C-MYC and C-FOS after DOX treatment in a dose-dependent manner (higher DOX concentration and lower expression were observed) in the case of MCF7 tumor cells. Application of MagAlg–DOX induced return of values closer to the control levels (Figure 7). The effect of MagAlg–DOX on C-MYC expression in NIH3T3 cells was not as marked as in MCF7 cells because NIH3T3 are non-tumor cells, thus with lower response to DOX. For C-FOS expression after treatment in NIH3T3 cells, we observed strong upregulation after treatment with 0.5 μM DOX, but downregulation after treatment with 5 μM DOX. Apparently, the higher C-FOS expression after treatment with lower dose of DOX is a reaction to oxidation or other after treatment stress, since C-FOS is involved in the stress response. On the other hand, 5 μM DOX is probably too high and cells and their DNA are damaged and the process of nucleic acid synthesis is inhibited. Importantly, NPs induced a return of values closer to control levels. C-FOS expression was increased after treatment with 5 μM MagAlg–DOX, probably due to oxidation or other stress, but this increase was not as high as in the case of 0.5 μM DOX only. C-FOS expression after treatment with 0.5 μM MagAlg–DOX was lower than when treated with 0.5 μM free DOX.


In vitro cytotoxicity analysis of doxorubicin-loaded/superparamagnetic iron oxide colloidal nanoassemblies on MCF7 and NIH3T3 cell lines.

Tomankova K, Polakova K, Pizova K, Binder S, Havrdova M, Kolarova M, Kriegova E, Zapletalova J, Malina L, Horakova J, Malohlava J, Kolokithas-Ntoukas A, Bakandritsos A, Kolarova H, Zboril R - Int J Nanomedicine (2015)

Fold change of C-MYC (A and B) and C-FOS (C and D) expression on MCF7 cells (left graphs) and NIH3T3 cells (right graphs) in the course of 0 μM, 0.5 μM, and 5 μM in DOX concentration and with effect of MagAlg–DOX nanocarrier.Notes: Data represent mean and standard error from three independent measurements. Positive (*) and negative (•) significance were determined using ANOVA and Dunnet post hoc test.Abbreviation: DOX, doxorubicin.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4321606&req=5

f7-ijn-10-949: Fold change of C-MYC (A and B) and C-FOS (C and D) expression on MCF7 cells (left graphs) and NIH3T3 cells (right graphs) in the course of 0 μM, 0.5 μM, and 5 μM in DOX concentration and with effect of MagAlg–DOX nanocarrier.Notes: Data represent mean and standard error from three independent measurements. Positive (*) and negative (•) significance were determined using ANOVA and Dunnet post hoc test.Abbreviation: DOX, doxorubicin.
Mentions: We observed downregulation of C-MYC and C-FOS after DOX treatment in a dose-dependent manner (higher DOX concentration and lower expression were observed) in the case of MCF7 tumor cells. Application of MagAlg–DOX induced return of values closer to the control levels (Figure 7). The effect of MagAlg–DOX on C-MYC expression in NIH3T3 cells was not as marked as in MCF7 cells because NIH3T3 are non-tumor cells, thus with lower response to DOX. For C-FOS expression after treatment in NIH3T3 cells, we observed strong upregulation after treatment with 0.5 μM DOX, but downregulation after treatment with 5 μM DOX. Apparently, the higher C-FOS expression after treatment with lower dose of DOX is a reaction to oxidation or other after treatment stress, since C-FOS is involved in the stress response. On the other hand, 5 μM DOX is probably too high and cells and their DNA are damaged and the process of nucleic acid synthesis is inhibited. Importantly, NPs induced a return of values closer to control levels. C-FOS expression was increased after treatment with 5 μM MagAlg–DOX, probably due to oxidation or other stress, but this increase was not as high as in the case of 0.5 μM DOX only. C-FOS expression after treatment with 0.5 μM MagAlg–DOX was lower than when treated with 0.5 μM free DOX.

Bottom Line: For proper analysis and understanding of cell behavior after administration of MagAlg-DOX compared with free DOX, a complex set of in vitro tests, including production of reactive oxygen species, comet assay, cell cycle determination, gene expression, and cellular uptake, were utilized.It was found that the cytotoxic effect of MagAlg-DOX system is delayed compared to free DOX in both cell lines.We discovered that nanoparticles can attenuate or even inhibit the effect of DOX, particularly in the tumor MCF7 cell line.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biophysics, Institute of Translation Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic.

ABSTRACT
One of the promising strategies for improvement of cancer treatment is based on magnetic drug delivery systems, thus avoiding side effects of standard chemotherapies. Superparamagnetic iron oxide (SPIO) nanoparticles have ideal properties to become a targeted magnetic drug delivery contrast probes, named theranostics. We worked with SPIO condensed colloidal nanocrystal clusters (MagAlg) prepared through a new soft biomineralization route in the presence of alginate as the polymeric shell and loaded with doxorubicin (DOX). The aim of this work was to study the in vitro cytotoxicity of these new MagAlg-DOX systems on mouse fibroblast and breast carcinoma cell lines. For proper analysis and understanding of cell behavior after administration of MagAlg-DOX compared with free DOX, a complex set of in vitro tests, including production of reactive oxygen species, comet assay, cell cycle determination, gene expression, and cellular uptake, were utilized. It was found that the cytotoxic effect of MagAlg-DOX system is delayed compared to free DOX in both cell lines. This was attributed to the different mechanism of internalization of DOX and MagAlg-DOX into the cells, together with the fact that the drug is strongly bound on the drug nanocarriers. We discovered that nanoparticles can attenuate or even inhibit the effect of DOX, particularly in the tumor MCF7 cell line. This is a first comprehensive study on the cytotoxic effect of DOX-loaded SPIO compared with free DOX on healthy and cancer cell lines, as well as on the induced changes in gene expression.

No MeSH data available.


Related in: MedlinePlus