Abstract
Development of tumour that is resistant to chemotherapeutics and synthetic drugs, coupled with their life-threatening side effects and the adverse effects of surgery and hormone therapies, led to increased research on probiotics’ anticancer potentials. The current study investigated the potential of live, heat-killed cells (HKC) and the cytoplasmic fractions (CF) of Enterococcus faecalis and Staphylococcus hominis as anti-breast cancer agents. MCF-7 cell line was treated with 25, 50, 100 and 200 μg/mL each of live, HKC and CF of the bacteria; and cytotoxicity was evaluated for 24, 48 and 72 h using MTT assay. The morphological features of the treated cells were examined by fluorescence microscopy. The stage of cell cycle arrest and apoptosis were quantified by flow cytometry. The bacterial effect on non-malignant breast epithelial cell line, MCF-10A, was assessed using MTT assay for 24, 48 and 72 h. All the three forms of the bacteria caused a significant decrease in MCF-7 (up to 33.29%) cell proliferation in concentration- and time-dependent manner. Morphological features of apoptosis like cell death, cell shrinkage and membrane blebbing were observed. Flow cytometry analyses suggested that about 34.60% of treated MCF-7 was undergoing apoptosis. A strong anti-proliferative activity was efficiently induced through sub-G1 accumulation (up to 83.17%) in treated MCF-7 and decreased number in the G0/G1 phase (74.39%). MCF-10A cells treated with both bacteria showed no significant difference with the untreated (>90% viability). These bacteria can be used as good alternative nutraceutical with promising therapeutic indexes for breast cancer because of their non-cytotoxic effects to normal cells.
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References
Biffi A, Coradini D, Larsen R, Riva L, Di Fronzo G (1997) Antiproliferative effect of fermented milk on the growth of a human breast cancer cell line. Nutr Cancer 28(1):93–99. doi:10.1080/01635589709514558
Choi SS, Kim Y, Han KS, You S, Oh S, Kim SH (2006) Effects of lactobacillus strains on cancer cell proliferation and oxidative stress in vitro. Lett Appl Microbiol 42:452–458
Cooper GM (2000) The cell: a molecular approach, 2nd edn. Boston University, Sunderland
De Moreno de LeBlanc A, Matar C, Leblanc N, Perdigon G (2005) Effect of milk fermented by Lactobacillus helveticus R389 on a murine. Breast Cancer Model 7(4):477–486
Di-Francia R, Stefania RS, Valente D, Del Buono A, Pugliese S, Cecere S, Berretta M. (2013) Current strategies to minimize toxicity of oxaliplatin : selection of pharmacogenomic panel tests. Anti-Cancer Drugs. 2:1–11. doi:10.1097/CAD.0000000000000002
Fotakis G, Timbrell JA (2006) In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett 160:171–177. doi:10.1016/j.toxlet.2005.07.001
Janghorbani M, Amini M, Willett WC, Gouya MM, Delavari A (2007) First nationwide survey of prevalence of overweight, underweight, and abdominal obesity in Iranian adults. Obesity 15(11):2797–2808
Kim JY, Woo HJ, Kim Y, Kim KH, Lee HJ (2003) Cell cycle dysregulation induced by cytoplasm of Lactococcus lactis ssp. lactis in SNUC2A, a colon cancer cell line. Nutr Cancer 46(2):197–201. doi:10.1207/S15327914NC4602
Lay MM, Karsani SA, Malek SNA (2014) Ethanone-induced cell cycle arrest in G1/G0 in HT-29 cells human colon adenocarcinoma cells. Int J Mol Sci 15:468–483. doi:10.3390/ijms15010468
Li J, Huang R, Yao G, Ye M, Wang H, Pan Y, Xiao J (2014) Synthesis and biological evaluation of novel aniline-derived asiatic acid derivatives as potential anticancer agents. Eur J Med Chem 86:175–188. doi:10.1016/j.ejmech.2014.08.003
Liu C-F, Pan T-M (2010) In vitro effects of lactic acid bacteria on cancer cell viability and antioxidant activity. J Food Drug Anal 18(2):77–86
Liu T, Chen C, Yiin S, Chen C, Cheng J, Shih M, Chern C (2006) Molecular mechanism of cell cycle blockage of hepatoma SK-Hep-1 cells by Epimedin C through suppression of mitogen-activated protein kinase activation and increased expression of CDK inhibitors p21 Cip1 and p27 Kip1. Food Chem Toxicol. 44:227–235. doi:10.1016/j.fct.2005.07.003
Nami Y, Abdullah N, Haghshenas B, Radiah D, Rosli R, Khosroushahi AY (2014) A newly isolated probiotic Enterococcus faecalis strain from vagina microbiota enhances apoptosis of human cancer cells. J Appl Microbiol 117:498–508. doi:10.1111/jam.12531
Roberts KM, Rosen A, Casciola-Rosen LA (2004) Methods for inducing apoptosis. Methods Mol Med 102:115–128. doi:10.1385/1-59259-805-6:115
Sato Y, Itoh F, Hareyama M et al (1999) Does cytoplasmic cyclin D1 regulate the cell cycle directly? J Gastroenterol 34:547–548
Shadeo A, Lam WL (2006) Comprehensive copy number profiles of breast cancer cell model genomes. Breast Cancer Res 8(1):R9. doi:10.1186/bcr1370
Subhashini S, Meignanalakshmi S (2013) In vitro studies on adhesion and the effect of cytotoxicity of Bifidobacterium spp. using cell lines. Eur Sci J 9(18):311–326
Thompson A, Brennan K, Cox A, Gee J, Harcourt D, Harris A, Steel M (2008) Evaluation of the current knowledge limitations in breast cancer research : a gap analysis. Breast Cancer Res. 10(R26):1–25. doi:10.1186/bcr1983
Wang L, Wu J, Lu J, Ma R, Sun D, Tang J (2014) Regulation of the cell cycle and PI3K/Akt/mTOR signaling pathway by tanshinone I in human breast cancer cell lines. Mol Med Rep. 1–9. doi:10.3892/mmr.2014.2819
WHO (2013) Latest world cancer statistics. International Agency for Research on Cancer, World Health Organization. Lyon/Geneva. Retrieved from http://www.iarc.fr/en/media-centre/pr/2013/pdfs/pr223_E.pdf
Acknowledgments
This project was supported by Fundamental Research Grants, Ministry of Higher Education, Malaysia (No. 5524244). We also acknowledge the invaluable assistance of Dr. Yeap Swee Keong of the Institute of BioScience, Universiti Putra Malaysia, Serdang, Malaysia, in the flow cytometry analysis and the data interpretation. We are also thankful to Associate Professor Dr. Noorjahan Banu Mohamed Alitheen, Universiti Putra, Malaysia, for allowing us to use her laboratory for some part of the study.
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Hassan, Z., Mustafa, S., Rahim, R.A. et al. Anti-breast cancer effects of live, heat-killed and cytoplasmic fractions of Enterococcus faecalis and Staphylococcus hominis isolated from human breast milk. In Vitro Cell.Dev.Biol.-Animal 52, 337–348 (2016). https://doi.org/10.1007/s11626-015-9978-8
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DOI: https://doi.org/10.1007/s11626-015-9978-8