Abstract
Objective
The CagA (cytotoxin-related gene A, CagA) protein is an important factor for the pathogenicity of Helicobacter pylori (H. pylori). Although H. pylori has previously been shown to activate the NLRP3 inflammasome, it remains unclear what role CagA plays in this process. In the current study, we aimed to investigate the effect of CagA on NLRP3 activation and how it is linked to gastric cancer cell migration and invasion.
Methods
CagA positive H. pylori strain (Hp/CagA+) and CagA gene knockout mutant (Hp/ΔCagA) infected and the pcDNA3.1/CagA plasmid transfected gastric epithelial cell lines, respectively. The morphological alterations of cells under a microscope; the NLRP3 inflammasome-related markers: NLRP3, caspase-1, and ASC protein levels were detected by Western blot, IL-1β and IL-18 levels were determined by ELISA; cell migration and invasion were determined by transwell assay; and the pyroptosis levels and intracellular ROS were determined by flow cytometry analysis. Then, pretreated with 5 mM NAC for 2 h and subsequently transfected with the pcDNA3.1/CagA plasmid for 48 h, the effects of NAC pretreatment on CagA-induced NLRP3 inflammasome-related markers expression and cell pyroptosis were examined, finally assessed the effect of CagA on migration and invasion in NLRP3-silenced cells.
Results
We found that Hp/CagA+ strain infection and pcDNA3.1/CagA vector transfection result in NLRP3 inflammasome activation, generation of intracellular ROS, and increased invasion and migration of gastric cancer cells. Moreover, we found that ROS inhibition via NAC effectively blocks NLRP3 activation and pyroptosis. Silencing of NLRP3 reduces the effects of CagA on gastric cancer cell migration and invasion.
Conclusion
Our study shows that CagA can promote the invasion and migration of gastric cancer cells by activating NLRP3 inflammasome pathway. These findings provide novel insights into the mechanism of gastric cancer induction by H. pylori.
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The authors are grateful to the editor, the associate editor, and the reviewer.
Funding
The work was supported by National Natural Science Foundation of China (31660031, 31760328, 31960028), Project of Science and Technology of Guiyang (ZhuKeHe[2017]30-4), The Key Project of Science and Technology of Guizhou Province(QianKeHe JC [2020]1Z010), and the Central Government Guides Local Science and Technology Development Projects of Guizhou (Grant no. [2019] 4008).
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YX conceived and designed the experiments, XYZ, YZ, YL, LC and DC performed the experiments and analyzed the data. QW, LYB and JJZ gave intellectual advice and revised the manuscript. YX and XYZ wrote the paper.
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The project was approved by the Ethic Committee of The Guizhou Medical University.
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Zhang, X., Li, C., Chen, D. et al. H. pylori CagA activates the NLRP3 inflammasome to promote gastric cancer cell migration and invasion. Inflamm. Res. 71, 141–155 (2022). https://doi.org/10.1007/s00011-021-01522-6
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DOI: https://doi.org/10.1007/s00011-021-01522-6