Fe-Blf Nanoformulation Targets Survivin to Kill Colon Cancer Stem Cells and Maintains Absorption of Iron, Calcium and Zinc

Abstract

Aim: To validate the anticancer efficacy of alginate-enclosed, chitosan-conjugated, calcium phosphate, iron-saturated bovine lactoferrin (Fe-bLf) nanocarriers/nanocapsules (NCs) with improved sustained release and ability to induce apoptosis by downregulating survivin, as well as cancer stem cells. Materials & methods: The stability, nanotoxicity of the modified nanoformulation was evaluated and their anticancer efficacy was re-examined. Their mechanism of internalization was studied and we identified the role of various miRNAs in absorption of these NCs/iron in various body parts of mice. We determined the effect of these NCs on survivin, stem cell markers, red blood cell count, iron, calcium and zinc concentration in mice, determined the antiangiogenic properties of these NCs and studied their effect on cancer stem-like cells. Results: Spherical NCs (396.1 ± 27.2 nm) exceedingly reduced viability of Caco-2 cells (32 ± 2.83%). The NCs also showed effective internalization and reduction of cancer stem cell markers in triple-positive CD133, survivin and CD44 cancer stem-like cells. Mice treated with the NCs showed no nanotoxicity and did not develop any tumors in xenograft colon cancer models. We found that the serum iron, zinc and calcium absorption were increased. DMT1, LRP, transferrin and lactoferrin receptors were responsible for internalization of the NCs. Different miRNAs were responsible for iron regulation in different organs. Interestingly, NCs inhibited survivin and its different isoforms. Conclusion: Our results confirmed that NCs internalized and changed the expression of selected miRNAs that further enhanced their uptake. The NCs activated both extrinsic, as well as intrinsic apoptotic pathways to induce apoptosis by targeting survivin in cancer cells and cancer stem cells, without inducing any nonspecific nanotoxicity. Apart from inhibiting angiogenesis and stem cell markers, NCs also maintained iron and calcium levels.

Original submitted 4 May 2014; Revised submitted 25 June 2014

Keywords::

• absorption
• Fe-bLf
• iron metabolism
• miRNA
• nanocarriers/nanocapsules
• nanotoxicity
• oral delivery
• stem cell
• survivin
• xenograft

Acknowledgements

The authors would like to thank N Branson for his assistance with animal work and for his technical support and guidance with the animal experimental procedures. The authors also thank the upper animal house staff, in particular T Thorpe, A Cooper and B Newell for providing help in handling the animals.

Financial & competing interests’ disclosure

The authors would like to thank the Australia–India Strategic Research Fund (AISRF) and National Health and Medical Research Council (NHMRC) for financial support. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all animal experimental investigations.

Additional information

Funding

The authors would like to thank the Australia–India Strategic Research Fund (AISRF) and National Health and Medical Research Council (NHMRC) for financial support. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.