Bacteroides fragilis Toxin Induces Sequential Proteolysis of E-Cadherin and Inflammatory Response in Mouse Intestinal Epithelial Cell Line
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and BFT Treatment
2.2. Bacterial Strains and Collection of Bacterial Supernatants
2.3. Western Blot Analysis
2.4. Immunofluorescence Staining and Confocal Microscopy
2.5. RNA Extraction, Reverse Transcription PCR, and DNA Sequencing
2.6. γ-Secretase and Proteasome Inhibitor Study
2.7. Evaluation of Cellular Proliferation
2.8. RayBio® Inflammation Antibody Array
2.9. Enzyme-Linked Immunosorbent Assay (ELISA) Analysis
2.10. Statistical Analysis
3. Results
3.1. BFT Induces Loss of E-Cadherin in Murine Epithelial CMT93 Cells
3.2. BFT Induces E-Cadherin Ectodomain Cleavage and Degradation of Intracellular Domain in CMT93 Cells
3.3. CMT93 Cells Resynthesize E-Cadherin and Undergo Proliferation After BFT Treatment
3.4. γ-Secretase Cleaves E-Cadherin Intracellular Domain
3.5. Proteasome Degrades E-Cadherin Cytoplasmic Fragment in CMT93 Cells
3.6. BFT Induces Secretion of CXCL1 and CXCL5
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
APC | Adenomatous polyposis coli |
BFT | Bacteroides fragilis toxin |
BHIA | Brain heart infusion agar |
BHIB | Brain heart infusion broth |
BSA | Bovine serum albumin |
cDNA | Complementary deoxyribose nucleic acid |
ELISA | Enzyme-linked immunosorbent assay |
ETBF | Enterotoxigenic Bacteroides fragilis |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
G-CSF | Granulocyte-colony stimulating factor |
GSK-3β | Glycogen synthase kinase-3 beta |
IL | Interleukin |
KC | Keratinocyte chemoattractant |
LIX | Lipopolysaccharide-induced CXC chemokine |
MCP-1 | Monocyte chemoattractant protein-1 |
MIP-1γ | Macrophage inflammatory protein-1 gamma |
MMP | Matrix metalloprotease |
mRNA | Messenger ribose nucleic acid |
NF-κB | Nuclear factor-kappa-light-chain-enhancer of activated B cells |
NTBF | Non-toxigenic Bacteroides fragilis |
rETBF | Recombinant enterotoxigenic Bacteroides fragilis |
rNTBF | Recombinant non-toxigenic Bacteroides fragilis |
S/N | Supernatant |
SEM | Standard error of the mean |
SFM | Serum-free medium |
STAT3 | Signal transducer and activator of transcription 3 |
sTNFR1 | Soluble tumor necrosis factor receptor 1 |
TGF-β | Transforming growth factor-beta |
Th17 | T helper 17 |
TNF-α | Tumor necrosis factor-alpha |
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Kim, W.-S.; Hwang, S.; Gwon, S.-Y.; Jo, M.; Yoo, S.-H.; Hong, J.; Jang, H.-N.; Hong, J.-E.; Kang, D.-H.; Yun, M.; et al. Bacteroides fragilis Toxin Induces Sequential Proteolysis of E-Cadherin and Inflammatory Response in Mouse Intestinal Epithelial Cell Line. Microorganisms 2025, 13, 781. https://doi.org/10.3390/microorganisms13040781
Kim W-S, Hwang S, Gwon S-Y, Jo M, Yoo S-H, Hong J, Jang H-N, Hong J-E, Kang D-H, Yun M, et al. Bacteroides fragilis Toxin Induces Sequential Proteolysis of E-Cadherin and Inflammatory Response in Mouse Intestinal Epithelial Cell Line. Microorganisms. 2025; 13(4):781. https://doi.org/10.3390/microorganisms13040781
Chicago/Turabian StyleKim, Woo-Seung, Soonjae Hwang, Sun-Yeong Gwon, Minjeong Jo, Sang-Hyeon Yoo, Jiyun Hong, Ha-Neul Jang, Ju-Eun Hong, Da-Hye Kang, Miyong Yun, and et al. 2025. "Bacteroides fragilis Toxin Induces Sequential Proteolysis of E-Cadherin and Inflammatory Response in Mouse Intestinal Epithelial Cell Line" Microorganisms 13, no. 4: 781. https://doi.org/10.3390/microorganisms13040781
APA StyleKim, W.-S., Hwang, S., Gwon, S.-Y., Jo, M., Yoo, S.-H., Hong, J., Jang, H.-N., Hong, J.-E., Kang, D.-H., Yun, M., & Rhee, K.-J. (2025). Bacteroides fragilis Toxin Induces Sequential Proteolysis of E-Cadherin and Inflammatory Response in Mouse Intestinal Epithelial Cell Line. Microorganisms, 13(4), 781. https://doi.org/10.3390/microorganisms13040781