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Design and validation of an orally administrated active L. fermentum-L. acidophilus probiotic formulation using colorectal cancer Apc Min/+ mouse model

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Abstract

Probiotics have been shown to have beneficial properties in attenuating the risk of colorectal cancer (CRC) development. However, functional evidence to support such effects for some probiotic bacteria are relatively unknown. Here, we document a significant antioxidant, anti-proliferative and pro-apoptotic activities of Lactobacillus acidophilus ATCC 314 and Lactobacillus fermentum NCIMB 5221 on CRC cells, particularly when used in combination (La-Lf). Furthermore, a superior synergistic activity on the inhibition of tumor growth and modulation of cell proliferation and epithelial markers in the Apc Min/+ CRC mouse model was explored, based on the expression levels of Ki-67, E-cadherin, β-catenin, and cleaved caspase-3 (CC3) proteins. The anti-cancer activity of La-Lf co-culture was significantly enhanced in vitro with significant reduced proliferation (38.8 ± 6.9 %, P = 0.009) and increased apoptosis (413 RUL, P < 0.001) towards cancer cells, as well as significant protection of normal colon cell growth from toxic treatment (18.6 ± 9.8 %, P = 0.001). La-Lf formulation (1010cfu/animal/day) altered aspects of intestinal tumorigenesis by significantly reducing intestinal tumor multiplicity (1.7-fold, P = 0.016) and downregulating cellular proliferation markers, including β-catenin (P = 0.041) and Ki-67 (P = 0.008). In conclusion, La-Lf showed greater protection against intestinal tumorigenesis supporting a potential use as a biotherapeutic for the prevention of CRC.

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Acknowledgments

The authors would like to acknowledge a Canadian Institute of Health Research (CIHR) grant (MPO 64308) and grants from Micropharma Limited to Dr. Satya Prakash, a Fonds de Recherche du Québec–Santé (FRSQ) Doctoral Awards and a Faculty of Medicine George G. Harris Fellowship to Imen Kahouli. The authors thank the Comparative Medicine and Animal Resources Centre (McGill University) and Dr. Martin Lapointe for his assistance with histopathological analysis. The authors acknowledge the assistance of Dr. Anis Riahi (Department of Neurology, Military Hospital of Tunis, Tunisia), as well as Nadia R. Hindri (Clinical Biology Laboratory, Salah Azaiz Institute, Tunisia) in the study design and data interpretation.

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Correspondence to Satya Prakash.

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This study was funded by a Canadian Institute of Health Research (CIHR) grant (MPO 64308).

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Imen Kahouli declares that she has no conflict of interest. Dr. Meenakshi Malhotra declares that she has no conflict of interest. Susan Westfall declares that she has no conflict of interest. Dr. Moulay A. Alaoui-Jamali declares that he has no conflict of interest. Dr. Satya Prakash declares that he has no conflict of interest.

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All applicable international, national, and institutional guidelines for the use and care of animals were followed.

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Kahouli, I., Malhotra, M., Westfall, S. et al. Design and validation of an orally administrated active L. fermentum-L. acidophilus probiotic formulation using colorectal cancer Apc Min/+ mouse model. Appl Microbiol Biotechnol 101, 1999–2019 (2017). https://doi.org/10.1007/s00253-016-7885-x

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