Limits...
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: Apo-bLf induced significantly greater cytotoxicity and reduction in cell proliferation in both cancer cells showing a time and dose dependent effect.Both forms of bLf significantly reduced cell invasion in cancer cells.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.

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

Cytotoxicity and proliferation in MDA-MB-231, MCF-7 and MCF-10-2A cells following Apo-bLf and Fe-bLf treatments. Lactate dehydrogenase assay (LDH) results demonstrating cytotoxicity in cells after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (a), MCF-7 (b) and MCF-10-2A cells (e). CyQUANT® assay results represent cell proliferation levels after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (c) and MCF-7 cells (d) including high (20% FBS) and low (untreated) controls. Data represented as mean with + SEM (n = 6). * = p < 0.05 compared with untreated (0 nM) group, ** = p < 0.01 compared with untreated (0 nM). Statistical analysis performed using Student’s t-test
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig1: Cytotoxicity and proliferation in MDA-MB-231, MCF-7 and MCF-10-2A cells following Apo-bLf and Fe-bLf treatments. Lactate dehydrogenase assay (LDH) results demonstrating cytotoxicity in cells after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (a), MCF-7 (b) and MCF-10-2A cells (e). CyQUANT® assay results represent cell proliferation levels after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (c) and MCF-7 cells (d) including high (20% FBS) and low (untreated) controls. Data represented as mean with + SEM (n = 6). * = p < 0.05 compared with untreated (0 nM) group, ** = p < 0.01 compared with untreated (0 nM). Statistical analysis performed using Student’s t-test

Mentions: Apo-bLf was the most effective at inducing cell cytotoxicity with significant increases in cytotoxicity at concentrations of 20, 30 and 40 nM in MDA-MB-231 (133.05%, p = 0.003, 112.53%, p = 0.003 and 110.17%, p = 0.007, respectively) and MCF-7 cells (68.58%, p = 0.001, 90.68%, p = 0.00001 and 99.31%, p = 0.000006, respectively) after 48 h (Fig. 1a). Furthermore, Apo-bLf demonstrated significant toxicity after 24 h in MCF-7 cells at concentrations of 20 nM (25.93%, p = 0.02), 30 nM (40.09%, p = 0.001) and 40 nM (61.97%, p = 0.001) (Fig. 1b). Fe-bLf demonstrated significant increases in cytotoxicity in MDA-MB-231 cells at concentrations of 10, 20 and 30 nM (51.81%, p = 0.04, 61.76%, p = 0.01 and 42.01%, p = 0.048) after 48 h (Fig. 1a) yet no significant cytotoxicity in MCF-7 cells (Fig. 1b).Fig. 1


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)

Cytotoxicity and proliferation in MDA-MB-231, MCF-7 and MCF-10-2A cells following Apo-bLf and Fe-bLf treatments. Lactate dehydrogenase assay (LDH) results demonstrating cytotoxicity in cells after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (a), MCF-7 (b) and MCF-10-2A cells (e). CyQUANT® assay results represent cell proliferation levels after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (c) and MCF-7 cells (d) including high (20% FBS) and low (untreated) controls. Data represented as mean with + SEM (n = 6). * = p < 0.05 compared with untreated (0 nM) group, ** = p < 0.01 compared with untreated (0 nM). Statistical analysis performed using Student’s t-test
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig1: Cytotoxicity and proliferation in MDA-MB-231, MCF-7 and MCF-10-2A cells following Apo-bLf and Fe-bLf treatments. Lactate dehydrogenase assay (LDH) results demonstrating cytotoxicity in cells after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (a), MCF-7 (b) and MCF-10-2A cells (e). CyQUANT® assay results represent cell proliferation levels after 24 and 48 h Apo and Fe-bLf treatments in MDA-MB-231 (c) and MCF-7 cells (d) including high (20% FBS) and low (untreated) controls. Data represented as mean with + SEM (n = 6). * = p < 0.05 compared with untreated (0 nM) group, ** = p < 0.01 compared with untreated (0 nM). Statistical analysis performed using Student’s t-test
Mentions: Apo-bLf was the most effective at inducing cell cytotoxicity with significant increases in cytotoxicity at concentrations of 20, 30 and 40 nM in MDA-MB-231 (133.05%, p = 0.003, 112.53%, p = 0.003 and 110.17%, p = 0.007, respectively) and MCF-7 cells (68.58%, p = 0.001, 90.68%, p = 0.00001 and 99.31%, p = 0.000006, respectively) after 48 h (Fig. 1a). Furthermore, Apo-bLf demonstrated significant toxicity after 24 h in MCF-7 cells at concentrations of 20 nM (25.93%, p = 0.02), 30 nM (40.09%, p = 0.001) and 40 nM (61.97%, p = 0.001) (Fig. 1b). Fe-bLf demonstrated significant increases in cytotoxicity in MDA-MB-231 cells at concentrations of 10, 20 and 30 nM (51.81%, p = 0.04, 61.76%, p = 0.01 and 42.01%, p = 0.048) after 48 h (Fig. 1a) yet no significant cytotoxicity in MCF-7 cells (Fig. 1b).Fig. 1

Bottom Line: Apo-bLf induced significantly greater cytotoxicity and reduction in cell proliferation in both cancer cells showing a time and dose dependent effect.Both forms of bLf significantly reduced cell invasion in cancer cells.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.

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