Effect of miR-223-3p on cell pyroptosis in myelodysplastic syndrome and its mechanism via regulating the expression of NLRP3

Abstract

This study aimed to investigate the regulatory mechanism of the miR-223-3p/NLRP3 signaling axis in the progression of myelodysplastic syndrome (MDS). For this purpose, SKM-1 cells were transfected and three groups were set up according to different transfection protocols: si-NC group (NLRP3 silencing negative control plasmid), si-NLRP3 group (NLRP3 silencing plasmid), miR-223-3p mimic- NC group (miR-223-3p overexpressing negative control plasmid), miR-223-3p mimic group (miR-223- 3p overexpressing plasmid), miR-223-3p mimic+oe-NLRP3 group and miR-223-3p mimic+oe-NLRP3 group (NLRP3 silencing combined with miR-223-3p overexpressing plasmid). Normal bone marrow cells were used as the control. qRT-PCR was used to detect relative expressions of NLRP3 and miR- 223-3p; and Western blot to detect Ki67, Caspase-1, Gasdermin D, IL-1β, IL-18 and MMP-9 expressions. Cell proliferation detection used CCK-8 assay, cell cycle distribution detection adopted flow cytometry, and cell migration and invasion analyses relied on Transwell assay. Dual-luciferase reporter assay verified the relationship between NLRP3 and miR-223-3p. An animal experiment was finally conducted to confirm the results obtained in cells. Results showed that compared with normal bone marrow cells and K562 cells, there were significantly upregulated NLRP3 expression and upregulated expression of miR-223-3p in SKM-1 cells (all P<0.05). Compared with the si-NC group and mimic-NC group respectively, the si-NLRP3 group and miR-223-3p mimic group showed inhibited proliferation, blocked cells in the G0/M phase, reduced cells in S phase, inhibited cell invasion and migration, decreased expressions of Ki67, Caspase-1, Gasdermin D, IL-1β and IL-18, and MMP-9 (all P<0.05). NLRP3 was the direct target of miR-223-3p. Moreover, compared with the miR-223-3p mimic group, the miR-223-3p mimic+oe-NLRP3 group showed increased expressions of NLRP3, Ki67, Caspase-1, Gasdermin D, IL-1β, IL-18 and MMP-9, promoted proliferation, invasion and migration, and increased cells in S phase (all P<0.05). The animal test revealed that compared with the mimic-NC+ oe- NC group, miR-223-3p mimic+ oe-NC group showed reduced tumor volume and decreased Ki67 expression (both P<0.05); while compared with miR-223-3p mimic+ oe-NC group, miR-223-3p mimic+ oe-NLRP3 group had increased tumor volume and increased Ki67 expression (both P<0.05). It was concluded that overexpression of miR-223-3p can effectively inhibit the expression of NLRP3 and cell pyroptosis in MDS.

Pathogen and human NDPK-proteins promote AML cell survival via monocyte NLRP3-inflammasome activation

“…In addition, the association of sterile NLRP3 inflammasome activation with AML and MDS is also well accepted [10][11][12][13][14][15][16]. To date, the role of NLRP3 inflammasomes in myeloid malignancies has focused on classical pyroptotic pathways, where cells dying by pyroptosis release DAMPS that further activate inflammation via TLRs including TLR4 [13,15,17,[47][48][49]. However, our study has highlighted the potential of pathogen derived NDPKs acting as PAMPS to activate NLRP3 independently of either TLR4 or pyroptosis.…”

“…Although a novel concept in these diseases, the association between microbiota and solid cancers is becoming accepted [41][42][43][44][45] and the association of inflammation with cancer is well established [44,46]. In addition, the association of sterile NLRP3 inflammasome activation with AML and MDS is also well accepted [10][11][12][13][14][15][16]. To date, the role of NLRP3 inflammasomes in myeloid malignancies has focused on classical pyroptotic pathways, where cells dying by pyroptosis release DAMPS that further activate inflammation via TLRs including TLR4 [13,15,17,[47][48][49].…”

“…Several studies have implicated the NLRP3-inflammasome in AML progression [10][11][12]. Equally, the importance of NLRP3 activation in the pathogenesis of myelodysplastic syndromes (MDS) is well established [13][14][15][16]. The NLRP3 inflammasome (comprised of the NOD-, leucine-rich repeat (LRR)-and pyrin domain (PYD)-containing protein 3 (NLRP3), the adapter apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and the effector protease caspase-1) orchestrates Toll-like receptor 4 (TLR4) responses to both sterile and infection-related inflammation.…”