Evaluation of probiotic bacteria effect on expression of VDR, IL3 and NOD2 genes in Caco-2 cell line

Document Type : Full Research Paper

Authors

1 Department of Microbiology, Faculty of Basic Sciences, Qom Branch, Islamic Azad University, Qom, Iran

2 Department of Mesanchymal Stem Cell, Academic Center for Education, Culture and Research,Qom branch, Iran

Abstract

Probiotics are live and beneficial microorganisms that are known as a factor for prevention of infectious diseases and cancer. In this study, the effects of cytotoxicity of Lactobacillus plantarum extract on the colon cancer cell line (Caco-2) as well as the expression of VDR, IL3 and NOD2 genes were investigated.The supernatant of L. plantarum culture was collected. Then, the effects of cytotoxicity and fluidity of bacteria and Supernatant on Caco-2 cancer cell line were investigated using MTT method. Also, the expression of VDR, IL3 and NOD2 gene was evaluated using Real-Time PCR. Data were analyzed using one-way ANOVA. Investigation of the inhibitory effect of L. plantarum culture supernatant on Caco-2 cells and gene expression at different concentrations showed that 100 μL/mL for VDR, IL3 and NOD2 had the highest increase in gene expression significantly after 48 hours. The MTT test showed the highest lethality at a concentration of 100 μL/mL. Gene expression at 100 μL/mL increased significantly at 24, 48 and 72 h for VDR, 48 h for IL3 and 72 h for NOD2 compared to the control group (P <0.05).
 L. plantarum can be used to develop a new treatment strategy with high efficacy, low side effects, biologically safe and lower cost. Based on this, it is suggested to investigate the treatment and prevention of colon cancer using L. plantarum.

Keywords


1. Arnold, M., Abnet, C.C., Neale, R.E., Vignat, J., Giovannucci, E.L. and McGlynn, K.A.  (2020). Global burden of 5 major types of gastrointestinal cancer. Gastroenterology 159: 335-349. 
2. 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. Nutrition and Cancer 28: 93-99. 
3. Carnero, A. and Paramio, J.M. (2014). The PTEN/PI3K/AKT pathway in vivo, cancer mouse models. Frontiers in Oncology 4: 1-6. 
4. Dong, L.M., Potter, J.D., White, E., Ulrich, C.M., Cardon, L.R. and Peters, U. (2008). Genetic susceptibility to cancer: the role of polymorphisms in candidate genes. JAMA 299: 2423-2436. 
5. Gahyva, S.M. and Siqueira, J.F. (2005). Direct genotoxicity and mutagenicity of endodontic substances and materials as evaluated by two prokaryotic test systems. Journal of Applied Oral Science 13: 387-392.
6. Gigola, G., Carriere, P., Novoa Díaz, M.B., Perdigon, G., Zwenger, A.O. and Gentili, C. (2021). Survival effect of probiotics in a rat model of colorectal cancer treated with capecitabine. World Journal of Gastrointestinal Oncology 13: 1518-1531. 
7. Hirata, M., Bamba, T. and Hosoda, S. (1993). The human colon cancer cell line CaCo-2 produces secretory components during enterocytic differentiation. Japanese Journal of Gastroenterology 28: 528-534. 
8. Kruis, W. (2004). Antibiotics and probiotics in inflammatory bowel disease. Alimentary Pharmacology and Therapeutics 20: 75-78. 
9. Kuntz, S., Krieghoff-Henning, E., Kather, J.N., Jutzi, T., Höhn, J. and Kiehl, L. (2021). Gastrointestinal cancer classification and prognostication from histology using deep learning: Systematic review. European Journal of Cancer 155: 200-215.
10. Ma E.L., Choi, Y.J., Choi, J., Pothoulakis, C., Rhee, S.H.  and Im, E. (2010). The anticancer effect of probiotic Bacillus polyfermenticus on human colon cancer cells is mediated through ErbB2 and ErbB3 inhibition. International Journal of Cancer 127: 780-790. 
11. Sattari, S. and Ahmadizadeh, C. (2018). The study of expression of PTEN and AKT1 genes in co-culturing of HT29 colon cancer cell line with Streptococcus thermophilus. Feyz 22: 624-631.
12. Sevda, E.R., Kopara, A.T. and Kivance, M. (2015). Cytotoxic effects of various lactic acid bacteria on Caco-2 cells. Journal of Biology 39: 23-30. 
13. Shi, L.H., Balakrishnan, K., Thiagarajah, K., Mohd Ismail, N.I. and Yin, O.S. (2016). Beneficial properties of probiotics. Tropical Life Sciences Research 27: 73-90. 
14. Śliżewska, K., Markowiak-Kopeć P. and Śliżewska, W. (2020). The Role of probiotics in cancer prevention. Cancers (Basel) 13: 1-8. 
15.    Taverniti, V. and Guglielmetti, S. (2011). The immunomodulatory properties of probiotic microorganisms beyond their viability (ghost probiotics: proposal of paraprobiotic concept). Genes & Nutrition 6: 261-274.
16. Teame, T., Wang, A., Xie, M., Zhang, Z., Yang, Y. and Ding, Q. (2020). Paraprobiotics and postbiotics of probiotic Lactobacilli, their positive effects on the host and action mechanisms: A review. Frontiers in Nutrition 7: 1-7. 
17. Tripathy, A., Dash, J., Kancharla, S., Kolli, P., Mahajan, D. and Senapati, S. (2021). Probiotics: A promising candidate for management of colorectal cancer. Cancers (Basel) 13: 1-8. 
18. Vlasova, A.N., Kandasamy, S., Chattha, K.S., Rajashekara, G. and Saif, L.J.  (2016). Comparison of probiotic lactobacilli and bifidobacteria effects, immune responses and rotavirus vaccines and infection in different host species. Veterinary Immunology and Immunopathology 172: 72-84. 
19. Williams, N.T. (2010). Probiotics. American Journal of Health-System Pharmacy 67: 449-458. 
20. Wu, S., Yoon, S., Zhang, Y.G., Lu, R., Xia, Y.  and Wan, J. (2015). Vitamin D receptor pathway is required for probiotic protection in colitis. American Journal of Physiology Gastrointestinal Liver Physiology 309: 341-349. 
21. Yavari, M. and Ahmadizadeh, C. (2020). Effect of the cellular extract of co-cultured Lactobacillus Casei on BAX and human β-Defensin 2 genes expression in HT29 cells. The Horizon of Medical Sciences 26: 364-381.
22. Yue, Y., Ye, K., Lu, J., Wang, X., Zhang S. and Liu, L. (2020). Probiotic strain Lactobacillus plantarum YYC-3 prevents colon cancer in mice by regulating the tumour microenvironment. Biomedicine and Pharmacotherapy 127: 1-8.