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)

• Published in: International journal of peptide research and therapeutics 2020-12, Vol.26 (4), p.2619-2631
• Main Authors: Maghsood, Faezeh, Johari, Behrooz, Rohani, Mahdi, Madanchi, Hamid, Saltanatpour, Zohreh, Kadivar, Mehdi
• Format: Article
• Language: English
• Subjects: Animal Anatomy; Apoptosis; Biochemistry; Biomedical and Life Sciences; Colon cancer; Colorectal cancer; Cyclooxygenase-2; Gelatin; Gelatinase B; Histology; Life Sciences; Metastases; Metastasis; Molecular Medicine; Molecular weight; Morphology; mRNA; Pharmaceutical Sciences/Technology; Pharmacology/Toxicology; Polymer Sciences; Polysaccharides; Probiotics; Stem cells; Tissue inhibitor of metalloproteinase 1
• ISSN: 1573-3149; 1573-3904
• DOI: 10.1007/s10989-020-10049-z

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   100 kD, and HK-SON fractions ( p   100 kD fraction significantly up-regulated the production level of TIMP-1 ( p   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.