Title : The effect of bacteroides thetaiotaomicron extracellular vesicles and their supernatant on gene expression involved in epithelial –mesenchymal transition in HCT-116 Cell line
Objective: Human gastrointestinal tract microbiota plays roles in metabolism and interaction with the host cells through several mechanisms such as outer membrane vesicle (OMVs). This study aims to address the effects of Bacteroides thetaiotaomicron (B. thetaiotaomicron) OMVs and its supernatant on the expression of Cdh1, Cdh2, Snail2, Zeb1, and Zeb2 genes involved in Epithelial-Mesenchymal Transition in HCT-116 Cell line.
Materials and Methods: The supernatant was extracted using centrifugation. OMVs extraction was done using tris-ethylenediaminetetraacetic acid (EDTA)-sodium deoxycholate buffer treatment and centrifugation. Protein content and profile of the OMVs were assessed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS- PAGE) and nanodrop. Protein concentration, Morphology, and lipopolysaccharide (LPS) of OMVs were distinguished by scanning electron microscope (SEM). The livability of OMVs was measure by 3- (4,5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium Bromide method (MTT) in HCT-116 cells. Human colon cancer (HCT-116) cell line as the model of intestinal cancer cells was used to study the effects of B. thetaiotaomicron’s OMVs and supernatant.
Results: B. thetaiotaomicron’s OMVs and supernatant increased the expression of Cdh1 genes while reduced Cdh2 and Zeb2 and Snail2 expression. No significant effect was observed on the gene expression of Zeb1. OMVs had further roles than supernatant on gene expression.
Conclusion: While B. thetaiotaomicron’s OMVs showed no cytotoxic effects on human intestinal cells, their effects on Cdh1, Cdh2, Zeb1, Snail2, and Zeb2 genes. Based on the features and benefits that have been shown in various studies, with more scrutiny they could be considered as promising factors and new candidates for treatment. However, more research is needed to examine their effects in more detail.
Keywords: B. thetaiotaomicron, Epithelial –Mesenchymal Transition, Extracellular Vesicles.