Inhibitory effectand mechanism of Salmonella Typhi-outer membrane vesicles on the proliferation of colorectal cancer cells by regulating ferroptosis
In recent years,the tremendous potential of bacterial outer membrane vesicles(OMVs)in anti-tumor therapy has attracted wide attention.To investigate the effect of Salmonella enterica serovar Typhi outer membrane vesicles(S.Typhi-OMVs)on the proliferation of human colorectal cancer cell line HT-29 and the possible mechanism of action,the outer membrane vesicles of different bacteria were extracted by ultracentrifugation.Cell proliferation assay(CCK-8)was performed to detect the effect of each bacterial outer membrane vesicles on HT-29 cell viability.Transcriptome sequencing was used to analyze the changes of gene expression levels in HT-29 cells after S.Typhi-OMVs treatment.The content changes of ferroptosis related markers were detected with the corresponding kits.Quantitative real-time polymerase chain reaction and western blot were applied to detect the changes of mRNA and protein expression of related genes,respectively.The results showed that S.Typhi-OMVs had the most significant inhibitory effect on the proliferation of HT-29 among the six bacterial outer membrane vesicles,and the inhibitory effect showed concentration and time gradient dependence.RNA-seq revealed that ferroptosis may have occurred in HT-29.After S.Typhi-OMVs treatment,intracellular iron deposition occurred,oxidative products increased,and antioxidants decreased,which was consistent with the biochemical characteristics of ferroptosis.SAT 1 was the gene with the largest mRNA expression change.The p53-SAT1-ALOX15 is one of the signaling pathways of ferroptosis,and the mRNA and protein expressions of p53,SAT1 and ALOX15 were all increased after S.Typhi-OMVs treatment of HT-29.The above results suggest that S.Typhi-OMVs were able to inhibit the proliferation of colorectal cancer cells HT-29,and the mechanism might be related to the induction of ferroptosis in HT-29 through the p53-SAT 1-ALOX15 signaling pathway.
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