The pro-tumorigenic property of the NLRP3 inflammasome may be mor

The pro-tumorigenic property of the NLRP3 inflammasome may be more related to its pro-inflammatory activity associated with IL-1β and IL-18 release. Further studies will be required to clarify the exact function played by the NLRP3 inflammasome in the DDR pathway. The anti-proliferative and pro-apoptotic functions of NLRP3 have been reported both here and elsewhere [39, 40], but the proposed oncosuppressive activity of the NLRP3 inflammasome now requires confirmation at least in alternative models

of inflammation-induced cancer. In summary, we have shown that MSU-induced DNA damage activated the NLRP3 inflammasome in a priming-independent manner, supported oxidative stimulation of DDR, and promoted p53 activation and subsequent cell death. These new roles identified for the NLRP3 inflammasome in suppressing DNA repair and enhancing p53-mediated apoptosis of innate cells will open new avenues of research that clarify the role of NLRP3 in diseases associated with aberrant cell death. C57BL/6 mice were purchased from the Biological Resource Center (BRC, A*STAR, Singapore).

Nlrp3−/− mice were kindly provided by J. Tschopp (University of Lausanne, Switzerland) [7] and casp-1−/− mice were a generous gift from R. A. Flavell [41]. All experiments were conducted with age-matched mice (8–12 weeks of age), and all mutants were backcrossed to C57BL/6 background for at least ten generations. Animals were bred under specific pathogen-free conditions at the BRC (Singapore). Experiments were performed under the approval of the Institutional Animal Care & Use Committee in compliance with the Law and Guidelines Acalabrutinib mw for Animal Experiments Carnitine palmitoyltransferase II of the BRC, Singapore. BM-derived DCs (BMDCs) from 8- to 12-week-old C57BL/6 mice and Nlrp3−/− and casp-1−/− mice were prepared

as previously described [8]. Cells (1 × 106 cells/mL) were stimulated in complete medium (IMDM with 10% FBS) in 96-well plates (Corning) and exposed to MSU crystals (250 μg/mL, Alexis), silica (silicon dioxide, 250 μg/mL, Sigma), ultrapure LPS (1 μg/mL, Alexis), camptothecin (1 μM, Sigma), rotenone (10 μM, Sigma), and H2O2 (100 mM, Sigma) for the indicated times. MSU preparations were assayed using the limulus amebocyte lysate test and were endotoxin free. For radiation experiments, BMDCs were rested for 24 h before being subjected to 4 or 10 Gy of γ-radiation and harvested after 8 or 24 h. RNA was extracted from three biological replicates as previously described [8], and 8 μg total RNA was used for cRNA target preparation following the Affymetrix GeneChip expression analysis technical manual (Affymetrix, Santa Clara, CA, USA). Biotinylated cRNA (15 μg) was hybridized to 12 Affymetrix GeneChip Mouse Genome MOE430 2.0. using the one-cycle target-labeling kit according to the manufacturer’s instructions. Microarray analysis was performed using R language and Bioconductor software [42].

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