Ferdowsi University of Mashhad

Document Type : Research Articles

Authors

1 Biology Department, Faculty of Science, University of Maragheh, Maragheh, Iran

2 Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran

Abstract

This study aimed to investigate the cytotoxicity of a probiotic mixture on human breast cancer cell lines. To prepare the mixture, local probiotic bacteria were cultured, and the lyophilized supernatant was applied for downstream experiments. The antioxidant activity, total phenol content (TPC), and fatty acid composition of the bacterial supernatant (BS) were also measured. The possible cytotoxic/anti-proliferative effect of the probiotic mixture was accessed on both breast cancer cell lines at different concentrations using MTT assay. Furthermore, the apoptosis-inducing effects of the same mixture was studied by DAPI staining. The highest level of antioxidant activity and total phenol content (TPC) were detected for the BS at 3200 µg/ml. According to the GC–MS analysis, linoleic acid (37.40 %) and oleic acid (26.93 %) were identified as the major fatty acids of the BS. The MTT assay showed that the BS has anti-proliferative effects on MDA-MB-231 and MCF-7 cells in a time- and dose-dependent manner (IC50: 3200 μg/ml). The apoptosis-inducing effects of the mixture was confirmed in both cell lines through morphological analyses of the cells’ nucleoli, and the formation of apoptotic bodies. According to these experiments, cytotoxic effects and apoptosis-inducing potential were confirmed for the BS against two human breast cancer cell lines, including MDA-MB-231, and MCF-7. Hence, it could be considered as a suitable anti-cancer agent.

Keywords

Babushok V., Linstrom P. and Zenkevich I. (2011) Retention indices for frequently reported compounds of plant essential oils. Journal of Physical and Chemical Reference Data 40:043101.
Bahmani S., Azarpira N. and Moazamian E. (2019) Anti-colon cancer activity of Bifidobacterium metabolites on colon cancer cell line SW742. The Turkish Journal of Gastroenterology 30:835.
Benzie I. F. and Strain J. J. (1996) The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Analytical biochemistry 239:70-76.
Biffi A., Coradini D., Larsen R., Riva L. and Di Fronzo G. (1997) Antiproliferative effect of fermented milk on the growth of a human breast cancer cell line.
Bougnoux P., Hajjaji N., Maheo K., Couet C. and Chevalier S. (2010) Fatty acids and breast cancer: sensitization to treatments and prevention of metastatic re-growth. Progress in lipid research 49:76-86.
Bray F., Ferlay J., Soerjomataram I., Siegel R. L., Torre L. A. and Jemal A. (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians 68:394-424.
Di Francia R., Siesto R. S., Valente D., Del Buono A., Pugliese S., Cecere S., and et al. (2013) Current strategies to minimize toxicity of oxaliplatin: selection of pharmacogenomic panel tests. Anti-cancer drugs 24:1069-1078.
Elmore S. (2007) Apoptosis: a review of programmed cell death. Toxicologic pathology 35:495-516.
Haghshenas B., Nami Y., Abdullah N., Radiah D., Rosli R. and Khosroushahi A. Y. (2014) Anti-proliferative effects of Enterococcus strains isolated from fermented dairy products on different cancer cell lines. Journal of functional foods 11:363-374.
Hassan Z., Mustafa S., Rahim R. A. and Isa N. M. (2016) Anti-breast cancer effects of live, heat-killed and cytoplasmic fractions of Enterococcus faecalis and Staphylococcus hominis isolated from human breast milk. In Vitro Cellular & Developmental Biology-Animal 52:337-348.
Irecta-Nájera C. A., del Rosario Huizar-López M., Casas-Solís J., Castro-Félix P. and Santerre A. (2017) Protective effect of Lactobacillus casei on DMH-induced colon carcinogenesis in mice. Probiotics and antimicrobial proteins 9:163-171.
Kadirareddy R. H., Vemuri S. and Palempalli U. (2016) Probiotic Conjugated Linoleic Acid Mediated Apoptosis in Breast Cancer Cells by Downregulation of NFκB. Asian Pacific journal of cancer prevention: APJCP 17:3395-3403.
Lee N.-K., Han K. J., Son S.-H., Eom S. J., Lee S.-K. and Paik H.-D. (2015) Multifunctional effect of probiotic Lactococcus lactis KC24 isolated from kimchi. LWT-Food Science and Technology 64:1036-1041.
Malik S. S., Saeed A., Baig M., Asif N., Masood N. and Yasmin A. (2018) Anticarcinogenecity of microbiota and probiotics in breast cancer. International journal of food properties 21:655-666.
Maroof H., Hassan Z. M., Mobarez A. M. and Mohamadabadi M. A. (2012) Lactobacillus acidophilus could modulate the immune response against breast cancer in murine model. Journal of clinical immunology 32:1353-1359.
Modarressi R. E.-V., Daneshvar M., Beigom M. and Mobasheri E. M. (2014) Lactobacillus acidophilus and Lactobacillus crispatus culture supernatants downregulate expression of cancer-testis genes in the MDA-MB-231 cell line. Asian Pacific Journal of Cancer Prevention 15:4255-4259.
Motevaseli E., Dianatpour A. and Ghafouri-Fard S. (2017) The role of probiotics in cancer treatment: emphasis on their in vivo and in vitro anti-metastatic effects. International journal of molecular and cellular medicine 6:66.
Nami Y., Abdullah N., Haghshenas B., Radiah D., Rosli R. and Khosroushahi A. Y. (2014) Probiotic potential and biotherapeutic effects of newly isolated vaginal Lactobacillus acidophilus 36YL strain on cancer cells. Anaerobe 28:29-36.
Nami Y., Haghshenas B., Haghshenas M., Abdullah N. and Yari Khosroushahi A. (2015) The prophylactic effect of probiotic Enterococcus lactis IW5 against different human cancer cells. Frontiers in microbiology 6:1317.
Nazir Y., Hussain S. A., Abdul Hamid A. and Song Y. (2018) Probiotics and their potential preventive and therapeutic role for cancer, high serum cholesterol, and allergic and HIV diseases. BioMed research international 2018.
Nouri Z., Neyazi N., Modarressi M. H., Karami F., Abedin-Do A., Taherian-Esfahani Z., Ghafouri-Fard S. and Motevaseli E. (2018) Down-regulation of TSGA10, AURKC, OIP5 and AKAP4 genes by Lactobacillus rhamnosus GG and Lactobacillus crispatus SJ-3C-US supernatants in HeLa cell line. Klinická onkologie 31.
Rezaei P. F., Fouladdel S., Cristofanon S., Ghaffari S., Amin G. and Azizi E. (2011) Comparative cellular and molecular analysis of cytotoxicity and apoptosis induction by doxorubicin and Baneh in human breast cancer T47D cells. Cytotechnology 63:503.
Rezaei P. F., Fouladdel S., Hassani S., Yousefbeyk F., Ghaffari S. M., Amin G. and et al. (2012) Induction of apoptosis and cell cycle arrest by pericarp polyphenol-rich extract of Baneh in human colon carcinoma HT29 cells. Food and Chemical Toxicology 50:1054-1059.
Singh R., Letai A. and Sarosiek K. (2019) Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins. Nature Reviews Molecular Cell Biology 20:175-193.
Singleton V. L., Orthofer R. and Lamuela-Raventós R. M. 1999. [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. In Methods in enzymology. Vol. 299. Elsevier. 152-178.
Torella J. P., Ford T. J., Kim S. N., Chen A. M., Way J. C. and Silver P. A. (2013) Tailored fatty acid synthesis via dynamic control of fatty acid elongation. Proceedings of the National Academy of Sciences 110:11290-11295.
Wang Y., Wu Y., Wang Y., Xu H., Mei X., Yu D., and et al. (2017) Antioxidant properties of probiotic bacteria. Nutrients 9:521.
Yu A.-Q. and Li L. (2016) The potential role of probiotics in cancer prevention and treatment. Nutrition and cancer 68:535-544.