Abstract:Objective: To investigate the effects of caspase1 (CASP1) on the cell cycle and pyroptosis of human acute myeloid leukemia (AML) cells THP-1. Methods: THP-1 cells were cultured and divided into the control group (normal THP-1 cells), pcDNA3.1-null group (negative control for overexpression of CASP1, transfected with 5.0 μg/mL pcDNA3.1-null plasmid into THP-1 cells for 24 h), and pcDNA3.1-CASP1 group (overexpression of CASP1, transfected with 5.0 μg/ml pcDNA3.1-CASP1 plasmid into THP-1 cells for 24 h). Cell proliferation activity was detected by CCK-8 assay. Apoptosis and cell cycle were detected by flow cytometry. CASP1, interleukin (IL)-1β, and IL-18 mRNA expression levels were detected by qRT-PCR. Cell proliferation-related proteins Ki67, proliferating cell nuclear antigen (PCNA), cyclin D1, NOD-like receptor (NLR)-like receptor pyrin domain-containing protein 3 (NLRP3), apoptosis-associated speck-like protein (ASC), CASP1, cleaved-caspase 1 (cleaved-CASP1), IL-1β, IL-18, and cleaved-Gasdermin D were detected by Western blot. Results: Compared with the control group, there were no significant differences in cell proliferation activity, apoptosis, and cell cycle changes in the pcDNA3.1-null group (P>0.05). Compared with the control group, cell proliferation activity was reduced, G0/G1 cell cycle was blocked, and the relative expression levels of Ki67, PCNA, and cyclin D1 were decreased (P<0.05) in the pcDNA3.1-CASP1 group. The relative expression levels of NLRP3, ASC, CASP1, cleaved-CASP1, IL-1β, IL-18, and cleaved-Gasdermin D were increased (P<0.05). Compared with the pcDNA3.1-null group, there were no significant differences in apoptosis in the pcDNA3.1-CASP1 group (P>0.05). However, cell proliferation activity was reduced, G0/G1 cell cycle was blocked, and the relative expression levels of Ki67, PCNA, and cyclin D1 were decreased (P<0.05). The relative expression levels of NLRP3, ASC, CASP1, cleaved-CASP1, IL-1β, IL-18, and cleaved-Gasdermin D were increased (P<0.05). Conclusion: Overexpression of CASP1 induces G0/G1 cell cycle arrest and NLRP3 inflammasome-mediated pyroptosis in AML cells THP-1.
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