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Cell-bound Exopolysaccharide Extract from Indigenous Probiotic Bacteria Induce Apoptosis in HT-29 cell-line.

ABSTRACT: Background and Objective:The aim of this study was to compare the cytotoxiceffects of local probiotic bacteria, including Lactobacillus paracasei, Lactobacillus brevis, while isolated from "Tarkhine" food and the induction of apoptosis in the HT-29 human colon adenocarcinoma cell line and normal fibroblasts. Methods:HT-29 and L-929 cell lines were treated with cell-bound exopolysaccharide extract (cb-EPS) from L. paracasei and L. brevis. The MTT assay was used to analyze cell viability. Cellular apoptosis was examined by flow cytometry and DNA fragmentation assay. Results:The cb-EPS from both probiotic bacteria prevented the proliferation of HT-29 colon cancer cells. In addi- tion, the cytotoxic and anti-proliferative effects of the exopolysaccharide extract from both bacteria in L-929 fibro- blasts were much lower than HT-29 cells. The induction of apoptosis in HT-29 cells was observed at 48h compared with 72h. It seems that the exopolysaccharides extracted from both bacteria have a greater effect on the induction of apoptosis at 48h. The cb-EPS of L. brevis showed more potent anti-proliferative and apoptotic properties than the cb- EPS of L. paracasei. The ladder pattern of DNA fragmentation confirmed the induction of apoptosis in cancer cells. Conclusion:The results of the MTT assay and apoptosis indicate that the induction of apoptosis by the exopolysac- charide from bacteria depends on the dose, time, and strain of bacteria. Further studies may contribute toward the understanding of using these probiotic bacteria as biological products to treat and prevent cancers.

SUBMITTER: Mojibi P

PROVIDER: S-EPMC6708565 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Cell-bound Exopolysaccharide Extract from Indigenous Probiotic Bacteria Induce Apoptosis in HT-29 cell-line.

Mojibi Parisa P Tafvizi Farzaneh F Bikhof Torbati Maryam M

Iranian journal of pathology 20181227 1

<h4>Background and objective</h4>The aim of this study was to compare the cytotoxiceffects of local probiotic bacteria, including <i>Lactobacillus paracasei, Lactobacillus brevis</i>, while isolated from "Tarkhine" food and the induction of apoptosis in the HT-29 human colon adenocarcinoma cell line and normal fibroblasts.<h4>Methods</h4>HT-29 and L-929 cell lines were treated with cell-bound exopolysaccharide extract (cb-EPS) from <i>L. paracasei</i> and <i>L. brevis</i>. The MTT assay was used ...[more]

PMID: 31531100

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Project description:BackgroundEpithelial cells in malignant conditions release DNA into the extracellular compartment. Cell free DNA of tumor origin may act as a ligand of DNA sensing mechanisms and mediate changes in epithelial-stromal interactions.AimsTo evaluate and compare the potential autocrine and paracrine regulatory effect of normal and malignant epithelial cell-related DNA on TLR9 and STING mediated pathways in HT-29 human colorectal adenocarcinoma cells and normal fibroblasts.Materials and methodsDNA isolated from normal and tumorous colonic epithelia of fresh frozen surgically removed tissue samples was used for 24 and 6 hour treatment of HT-29 colon carcinoma and HDF-? fibroblast cells. Whole genome mRNA expression analysis and qRT-PCR was performed for the elements/members of TLR9 signaling pathway. Immunocytochemistry was performed for epithelial markers (i.e. CK20 and E-cadherin), DNA methyltransferase 3a (DNMT3a) and NF?B (for treated HDF? cells).ResultsAdministration of tumor derived DNA on HT29 cells resulted in significant (p<0.05) mRNA level alteration in 118 genes (logFc?1, p?0.05), including overexpression of metallothionein genes (i.e. MT1H, MT1X, MT1P2, MT2A), metastasis-associated genes (i.e. TACSTD2, MACC1, MALAT1), tumor biomarker (CEACAM5), metabolic genes (i.e. INSIG1, LIPG), messenger molecule genes (i.e. DAPP, CREB3L2). Increased protein levels of CK20, E-cadherin, and DNMT3a was observed after tumor DNA treatment in HT-29 cells. Healthy DNA treatment affected mRNA expression of 613 genes (logFc?1, p?0.05), including increased expression of key adaptor molecules of TLR9 pathway (e.g. MYD88, IRAK2, NF?B, IL8, IL-1?), STING pathway (ADAR, IRF7, CXCL10, CASP1) and the FGF2 gene.ConclusionsDNA from tumorous colon epithelium, but not from the normal epithelial cells acts as a pro-metastatic factor to HT-29 cells through the overexpression of pro-metastatic genes through TLR9/MYD88 independent pathway. In contrast, DNA derived from healthy colonic epithelium induced TLR9 and STING signaling pathway in normal fibroblasts.

Dataset Information

Cell-bound Exopolysaccharide Extract from Indigenous Probiotic Bacteria Induce Apoptosis in HT-29 cell-line.

Publications

Cell-bound Exopolysaccharide Extract from Indigenous Probiotic Bacteria Induce Apoptosis in HT-29 cell-line.

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