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Molecular Mechanisms of RNPEP-Regulated Macrophage Pyrodeath Involved in the Biological Behaviour of Gastric Cancer Cells |
HUO Ya'nan, YANG Chen, GUO Shuli, et al |
Handan Central Hospital, Hebei Handan 056000, China |
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Abstract Objective: To investigate the effect of RNPEP on the biological behavior of gastric cancer cells and its possible mechanism.Methods: RNPEP mRNA levels in normal gastric tissues and gastric cancer tissues were detected by RT-qPCR. THP-1 cells were cultured and induced into M0 macrophages, which were divided into control group and RNPEP overexpression group. The expression levels of RNPEP, GSDME, NLRP3, IL-1β and IL-18 in macrophages were detected by Western blotting. Human gastric adenocarcinoma cell MGC-803 was co-cultured with M0 macrophages of the control and RNPEP overexpression groups, and divided into control group and RNPEP overexpression group. MGC-803 cells and M0 macrophages in the over-expressing RNPEP group were co-cultured, and IL-1β IgG1 homologous antibody and IL-1β IgG1 neutralizing antibody were added to the culture medium of the two groups, respectively, and they were divided into the RNPEP over-expression group and the inhibiting IL-1β group. The proliferation ability of MGC-803 gastric cancer cells was detected by EdU kit. The invasive ability of MGC-803 gastric cancer cells was detected by Transwell assay. The migration ability of MGC-803 gastric cancer cells was detected by scratch test.Results: Compared with normal tissues, mRNA level of RNPEP was increased in gastric cancer tissues (P<0.05). Compared with the control group, the expressions of RNPEP, GSDME, NLRP3, IL-1β, and IL-18 in macrophages of the RNPEP over-expression group were increased (P<0.05). Compared with the control group, the proliferation, migration and invasion ability of MGC-803 cells in the RNPEP over-expression group were increased (P<0.05). Compared with the RNPEP over-expression group, the proliferation, migration, and invasion ability of MGC-803 cells in IL-1β inhibition group were decreased (P<0.05).Conclusion:The secreted protein RNPEP can promote the biological behavior of gastric cancer cells by up-regulating pyroptosis of macrophages.
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