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Iron-free and iron-saturated bovine lactoferrin inhibit survivin expression and differentially modulate apoptosis in breast cancer.

Gibbons JA, Kanwar JR, Kanwar RK - BMC Cancer (2015)

Bottom Line: Iron binding, naturally occurring protein bovine lactoferrin (bLf) has attracted attention as a safe anti-cancer agent capable of inducing apoptosis.Apo-bLf induced significantly greater cytotoxicity and reduction in cell proliferation in both cancer cells showing a time and dose dependent effect.Key apoptotic molecules including p53, Bcl-2 family proteins, IAP members and their inhibitors were significantly modulated by both forms of bLf, though differentially in each cell line.

View Article: PubMed Central - PubMed

Affiliation: Nanomedicine - Laboratory for Immunology and Molecular Biomedical Research, Molecular and Medical Research Facility, School of Medicine, Faculty of Health, Deakin University, Geelong, Victoria, Australia. jgib@deakin.edu.au.

ABSTRACT

Background: Iron binding, naturally occurring protein bovine lactoferrin (bLf) has attracted attention as a safe anti-cancer agent capable of inducing apoptosis. Naturally, bLf exists partially saturated (15-20%) with Fe(3+) however, it has been demonstrated that manipulating the saturation state can enhance bLf's anti-cancer activities.

Methods: Apo-bLf (Fe(3+) free) and Fe-bLf (>90% Fe(3+) Saturated) were therefore, tested in MDA-MB-231 and MCF-7 human breast cancer cells in terms of cytotoxicity, proliferation, migration and invasion. Annexin-V Fluos staining was also employed in addition to apoptotic protein arrays and Western blotting to determine the specific mechanism of bLf-induced apoptosis with a key focus on p53 and inhibitor of apoptosis proteins (IAP), specifically survivin.

Results: Apo-bLf induced significantly greater cytotoxicity and reduction in cell proliferation in both cancer cells showing a time and dose dependent effect. Importantly, no cytotoxicity was detected in normal MCF-10-2A cells. Both forms of bLf significantly reduced cell invasion in cancer cells. Key apoptotic molecules including p53, Bcl-2 family proteins, IAP members and their inhibitors were significantly modulated by both forms of bLf, though differentially in each cell line. Most interestingly, both Apo-bLf and Fe-bLf completely inhibited the expression of survivin protein (key IAP), after 48 h at 30 and 40 nM in cancer cells.

Conclusions: The capacity of these forms of bLf to target survivin expression and modulation of apoptosis demonstrates an exciting potential for bLf as an anti-cancer therapeutic in the existing void of survivin inhibitors, with a lack of successful inhibitors in the clinical management of cancer.

No MeSH data available.


Related in: MedlinePlus

Effect on migration and invasion capacity of MDA-MB-231 and MCF-7 cells after treatment with bLf. Migration of MDA-MB-231 and MCF-7 (a) cells after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control migration. Invasion of MDA-MB-231 and MCF-7 cells (b) after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control invasion. * = p < 0.05 compared with 1% FBS group. Representative images (250X magnification) of invaded cells stained with crystal violet (c)
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Fig3: Effect on migration and invasion capacity of MDA-MB-231 and MCF-7 cells after treatment with bLf. Migration of MDA-MB-231 and MCF-7 (a) cells after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control migration. Invasion of MDA-MB-231 and MCF-7 cells (b) after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control invasion. * = p < 0.05 compared with 1% FBS group. Representative images (250X magnification) of invaded cells stained with crystal violet (c)

Mentions: As migration and invasion are key properties of cancer cells leading to metastases and secondary tumour sites within the body, assays were performed to determine the effects of bLf in regards to these properties. Migration assays indicated a significant reduction in the capacity of MDA-MB-231 and MCF-7 (Fig. 3a) cells to migrate through a porous membrane after treatment for 24 h with each Apo-bLf and Fe-bLf at concentrations of 5 and 10 nM. The greatest reduction in migration was observed with 10 nM Apo-bLf with 26.45% (p = 0.001) of MDA-MB-231 and 38.78% (p = 0.001) of MCF-7 cells migrating. Invasion assays (Fig. 3b and c) were performed with ECM. The same trend was observed in MDA-MB-231 cells with 29.85% (p = 0.001) of cells invading after treatment with 10 nM Apo-bLf. MCF-7 showed a large reduction with 10 nM Apo-bLf (50.00%, p = 0.003) however the greatest reduction in invasion was with 10 nM Fe-bLf treatments with 26.25% (p = 0.00008) invasion.Fig. 3


Iron-free and iron-saturated bovine lactoferrin inhibit survivin expression and differentially modulate apoptosis in breast cancer.

Gibbons JA, Kanwar JR, Kanwar RK - BMC Cancer (2015)

Effect on migration and invasion capacity of MDA-MB-231 and MCF-7 cells after treatment with bLf. Migration of MDA-MB-231 and MCF-7 (a) cells after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control migration. Invasion of MDA-MB-231 and MCF-7 cells (b) after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control invasion. * = p < 0.05 compared with 1% FBS group. Representative images (250X magnification) of invaded cells stained with crystal violet (c)
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4440599&req=5

Fig3: Effect on migration and invasion capacity of MDA-MB-231 and MCF-7 cells after treatment with bLf. Migration of MDA-MB-231 and MCF-7 (a) cells after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control migration. Invasion of MDA-MB-231 and MCF-7 cells (b) after bLf treatments for 24 h represented as a percentage of untreated (1% FBS) control invasion. * = p < 0.05 compared with 1% FBS group. Representative images (250X magnification) of invaded cells stained with crystal violet (c)
Mentions: As migration and invasion are key properties of cancer cells leading to metastases and secondary tumour sites within the body, assays were performed to determine the effects of bLf in regards to these properties. Migration assays indicated a significant reduction in the capacity of MDA-MB-231 and MCF-7 (Fig. 3a) cells to migrate through a porous membrane after treatment for 24 h with each Apo-bLf and Fe-bLf at concentrations of 5 and 10 nM. The greatest reduction in migration was observed with 10 nM Apo-bLf with 26.45% (p = 0.001) of MDA-MB-231 and 38.78% (p = 0.001) of MCF-7 cells migrating. Invasion assays (Fig. 3b and c) were performed with ECM. The same trend was observed in MDA-MB-231 cells with 29.85% (p = 0.001) of cells invading after treatment with 10 nM Apo-bLf. MCF-7 showed a large reduction with 10 nM Apo-bLf (50.00%, p = 0.003) however the greatest reduction in invasion was with 10 nM Fe-bLf treatments with 26.25% (p = 0.00008) invasion.Fig. 3

Bottom Line: Iron binding, naturally occurring protein bovine lactoferrin (bLf) has attracted attention as a safe anti-cancer agent capable of inducing apoptosis.Apo-bLf induced significantly greater cytotoxicity and reduction in cell proliferation in both cancer cells showing a time and dose dependent effect.Key apoptotic molecules including p53, Bcl-2 family proteins, IAP members and their inhibitors were significantly modulated by both forms of bLf, though differentially in each cell line.

View Article: PubMed Central - PubMed

Affiliation: Nanomedicine - Laboratory for Immunology and Molecular Biomedical Research, Molecular and Medical Research Facility, School of Medicine, Faculty of Health, Deakin University, Geelong, Victoria, Australia. jgib@deakin.edu.au.

ABSTRACT

Background: Iron binding, naturally occurring protein bovine lactoferrin (bLf) has attracted attention as a safe anti-cancer agent capable of inducing apoptosis. Naturally, bLf exists partially saturated (15-20%) with Fe(3+) however, it has been demonstrated that manipulating the saturation state can enhance bLf's anti-cancer activities.

Methods: Apo-bLf (Fe(3+) free) and Fe-bLf (>90% Fe(3+) Saturated) were therefore, tested in MDA-MB-231 and MCF-7 human breast cancer cells in terms of cytotoxicity, proliferation, migration and invasion. Annexin-V Fluos staining was also employed in addition to apoptotic protein arrays and Western blotting to determine the specific mechanism of bLf-induced apoptosis with a key focus on p53 and inhibitor of apoptosis proteins (IAP), specifically survivin.

Results: Apo-bLf induced significantly greater cytotoxicity and reduction in cell proliferation in both cancer cells showing a time and dose dependent effect. Importantly, no cytotoxicity was detected in normal MCF-10-2A cells. Both forms of bLf significantly reduced cell invasion in cancer cells. Key apoptotic molecules including p53, Bcl-2 family proteins, IAP members and their inhibitors were significantly modulated by both forms of bLf, though differentially in each cell line. Most interestingly, both Apo-bLf and Fe-bLf completely inhibited the expression of survivin protein (key IAP), after 48 h at 30 and 40 nM in cancer cells.

Conclusions: The capacity of these forms of bLf to target survivin expression and modulation of apoptosis demonstrates an exciting potential for bLf as an anti-cancer therapeutic in the existing void of survivin inhibitors, with a lack of successful inhibitors in the clinical management of cancer.

No MeSH data available.


Related in: MedlinePlus