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Anti-Proliferative and Anti-Metastatic Potential of High Molecular Weight Secretory Molecules From Probiotic Lactobacillus Reuteri Cell-Free Supernatant Against Human Colon Cancer Stem-Like Cells (Ht29-She) Publisher



Maghsood F1, 2 ; Johari B3 ; Rohani M4 ; Madanchi H5 ; Saltanatpour Z6 ; Kadivar M1
Authors
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Authors Affiliations
  1. 1. Department of Biochemistry, Pasteur Institute of Iran, P O Box: 1316943551, Tehran, Iran
  2. 2. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
  4. 4. Department of Microbiology, Pasteur Institute of Iran, Tehran, Iran
  5. 5. Department of Biotechnology, School of Medicine, Semnan University of Medical Sciences, Semnan, Iran
  6. 6. Stem Cell and Regenerative Medicine Center of Excellence, Tehran University of Medical Sciences, Tehran, Iran

Source: International Journal of Peptide Research and Therapeutics Published:2020


Abstract

Cancer stem cells population are necessary for cancerous invasion. Previous studies have indicated anti-tumor properties of probiotic Lactobacillus reuteri. At the current study, the effect of heat-killed sonicated (HK-SON) fraction and defined molecular weight secreted fractions from L. reuteri cell-free supernatant (CFS) on invasion and apoptosis levels of HT29-ShE cells was investigated. HT29-ShE cells were treated with a range of defined molecular weight fractions derived from CFS and HK-SON. A group of the cells was treated with uninoculated sterile bacterial growth media (MRS) as the negative control for 24 h. Afterward, real-time PCR analysis was done for investigating the relative production level of MMP-9, TIMP-1, and COX-2 mRNA. Matrigel-coated insert plates were used to evaluate cell invasion. The pro-apoptotic effect of CFS and HK-SON was determined using flowcytometry, and the gelatinolytic role of MMP-9 was analyzed via gelatin zymography. HK-SON treatment and defined fractions of CFS significantly decreased the cell invasion versus the MRS group (p < 0.05). Moreover, expression level and activity of MMP-9 was considerably reduced after processing with 10–50 kD, 50–100 kD, and > 100 kD, and HK-SON fractions (p < 0.05). Furthermore, crude CFS, and > 100 kD fraction significantly up-regulated the production level of TIMP-1 (p < 0.05) and HK-SON and > 100 kD fractions significantly increased the apoptosis percentage of HT29-ShE cells. Our results indicate the anti-metastatic and anti-proliferative properties of CFS derived from L. reuteri. Findings suggest that a secretory macromolecule (s) such as a polysaccharide, nucleic acid, or protein might exert these inhibitory effects on colon cancer stem-like cells. In conclusion, our results indicated that cell-free supernatant components can be used as the anti-metastatic agent. However, this study must be performed in experimental and in vivo models for further results. © 2020, Springer Nature B.V.
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