Abstract:
ObjectiveTo investigate the expression of NLRP3 inflammasome in gastric cancer(GC) and its effects on the proliferation, invasion and migration of cells, and the role of NLRP3-mediated caspase-1-dependent pyroptosis signaling pathway in the development of GC.
MethodsThe expression levels of NLRP3 and caspase-1 in human gastric epithelial cells(GES-1) and human gastric cancer cells(MKN45 and MGC803) were detected by qRT-PCR and Western blotting.After the expression level of NLRP3 was down-regulated, and the mRNA and protein expression levels of NLRP3 and caspase-1 were detected, the protein expression levels of GSDMD, IL-18 and IL-1β were detected using Western blotting.The content of IL-1β and IL-18 in supernatant were detected by ELISA, the pyroptosis was detected by LDH and flow cytometry, and the cell proliferation was detected by CCK-8 assay and cell cycle kit.Transwell and wound healing were used to detect the invasion and migration of cells.
ResultsThe expression levels of NLRP3 and caspase-1 in GC cells were higher than those in GES-1(P < 0.01).After the expression level of NLRP3 was downregulated, the release LDH rate and pyroptosis rate of LDH significantly decreased(P < 0.01), and the expression levels of GSDMD, IL-1β and IL-18 decreased(P < 0.01).Compared with the NC group, the cell proliferation was arrested in G1 phase(P < 0.01).The proliferation, invasion and migration of siNLRP3 cells were significantly reduced.
ConclusionsThe expression of NLRP3 in GC cells is high, and its expression affects the proliferation, invasion and migration of GC cells and expression of key proteins in caspase-1-dependent pyroptosis signaling pathway mediated by NLRP3.