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Porphyromonas gingivalis Lipopolysaccharide Damages Mucosal Barrier to Promote Gastritis-Associated Carcinogenesis

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Abstract

Background

Recent epidemiological studies suggested correlation between gastric cancer (GC) and periodontal disease.

Aims

We aim to clarify involvement of lipopolysaccharide of Porphyromonas gingivalis (Pg.), one of the red complex periodontal pathogens, in the GC development.

Methods

To evaluate barrier function of background mucosa against the stimulations, we applied biopsy samples from 76 patients with GC using a Ussing chamber system (UCs). K19-Wnt1/C2mE transgenic (Gan) mice and human GC cell-lines ± THP1-derived macrophage was applied to investigate the role of Pg. lipopolysaccharide in inflammation-associated carcinogenesis.

Results

In the UCs, Pg. lipopolysaccharide reduced the impedance of metaplastic and inflamed mucosa with increases in mRNA expression of toll-like receptor (TLR) 2, tumor necrosis factor (TNF) α, and apoptotic markers. In vitro, Pg. lipopolysaccharide promoted reactive oxidative stress (ROS)-related apoptosis as well as activated TLR2-β-catenin-signaling on MKN7, and it increased the TNFα production on macrophages, respectively. TNFα alone activated TLR2-β-catenin-signaling in MKN7, while it further increased ROS and TNFα in macrophages. Under coculture with macrophages isolated after stimulation with Pg. lipopolysaccharide, β-catenin-signaling in MKN7 was activated with an increase in supernatant TNFα concentration, both of which were decreased by adding a TNFα neutralization antibody into the supernatant. In Gan mice with 15-week oral administration of Pg. lipopolysaccharide, tumor enlargement with β-catenin-signaling activation were observed with an increase in TNFα with macrophage infiltration.

Conclusions

Local exposure of Pg. lipopolysaccharide may increase ROS on premalignant gastric mucosa to induce apoptosis-associated barrier dysfunction and to secrete TNFα from activated macrophages, and both stimulation of Pg. lipopolysaccharide and TNFα might activate TLR2-β-catenin-signaling in GC.

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Abbreviations

LPS:

Lipopolysaccharide

mUCs:

mini-Ussing chamber system

NAC:

N-acetyl-l-cysteine

Pg.:

Porphyromonas gingivalis

PG:

Peptidoglycan

uSS:

Updated Sydney system

qPCR:

Quantitative polymerase chain reaction

ROS:

Reactive oxygen species

shRNA:

Short hairpin RNA

TLR:

Toll-like receptor

TNFα:

Tumor necrosis factor α

TUNEL:

TdT-mediated dUTP nick end labeling

HRP:

Horse radish peroxidase

GC:

Gastric cancer

H. pylori :

Helicobacter pylori

post-eradication GC:

Gastric cancer after eradication treatment of H. pylori

PMA:

Phorbol 12-myristate 13-acetate

Mφ:

Macrophages

Gan mice:

K19-Wnt1/C2mE transgenic mice

IQR:

Interquartile range

SD:

Standard deviation

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Acknowledgments

We thank Biomedical Research Unit of Tohoku University Hospital for technical support, and we thank the Cancer Research Institute of Kanazawa University for kind gift of K19-Wnt1/C2mE mice.

Funding

This work was supported by JSPS KAKENHI grant number 19K08434, and the grant for Basic Research of the Japanese Society for Helicobacter research (2020).

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Authors and Affiliations

  1. Division of Gastroenterology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-cho Aoba-ku, Sendai, Miyagi, 981-8574, Japan

    Masayoshi Oriuchi, Sujae Lee, Kaname Uno, Koichiro Sudo, Keisuke Kusano, Naoki Asano, Shin Hamada, Waku Hatta, Tomoyuki Koike, Akira Imatani & Atsushi Masamune

  2. Division of Gastroenterology, Tohoku University Hospital, Sendai, Japan

    Kaname Uno

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  1. Masayoshi Oriuchi
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Correspondence to Kaname Uno.

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Supplementary file1 (TIF 3813 kb)

Supplementary Figure. Comprehensive analysis of h-Pg. LPS-stimulated apoptosis-related gene expressions in MKN7. The apoptosis antibody array demonstrated the induction of the proteins of apoptosis-signaling and TNF-signaling in MKN7 (N = 2: Int/pos = the ratio of signal of the target gene per those of positive control).

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Oriuchi, M., Lee, S., Uno, K. et al. Porphyromonas gingivalis Lipopolysaccharide Damages Mucosal Barrier to Promote Gastritis-Associated Carcinogenesis. Dig Dis Sci 69, 95–111 (2024). https://doi.org/10.1007/s10620-023-08142-6

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