Department of Pharmacology, College of Medicine, Institute of Health Sciences, Gyeongsang National University, 52727 Jinju, Korea.
Int J Mol Sci. 2020 May 8;21(9):3337. doi: 10.3390/ijms21093337.
The inflammasomes are reported to be associated with tumor progression. In our previous study, we determined that extracellular ATP enhances invasion and tumor growth by inducing inflammasome activation in a P2Y purinergic receptor 2 (P2YR)-dependent manner. However, it is not clear which inflammasome among the diverse complexes is associated with P2YR activation in breast cancer. Thus, in this study, we determined which inflammasome components are regulated by P2YR activation and are involved in tumor progression in breast cancer cells and radiotherapy-resistant (RT-R)-breast cancer cells. First, we found that NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3); NLR family caspase activation and recruitment domain (CARD) containing 4 (NLRC4); apoptosis-associated speck-like protein containing a CARD complex (ASC); and caspase-1 mRNA levels were upregulated in RT-R-MDA-MB-231 cells compared to MDA-MB-231 cells, whereas tumor necrosis factor-α (TNF-α) or ATP treatment induced NLRC4, ASC, and caspase-1 but not NLRP3 protein levels. Moreover, TNF-α or ATP increased protein levels of NLRC4, ASC, and caspase-1 in a P2YR-dependent manner in MDA-MB-231 and RT-R-MDA-MB-231 cells. In addition, P2YR activation by ATP induced the secretion of IL-1β and VEGF-A, as well as invasion, in MDA-MB-231 and RT-R-MDA-MB-231 cells, which was inhibited by NLRC4, ASC, and caspase-1 small interfering RNA (siRNA). Taken together, this report suggests that P2YR activation by ATP induces tumor invasion and angiogenesis through inflammasome activation, specifically by regulating the inflammasome components NLRC4, ASC, and caspase-1.
炎性小体被报道与肿瘤进展有关。在我们之前的研究中，我们确定细胞外 ATP 通过诱导 P2Y 嘌呤能受体 2 (P2YR) 依赖性的炎性小体激活，增强侵袭和肿瘤生长。然而，在乳腺癌中，哪种炎性小体与 P2YR 激活有关尚不清楚。因此，在这项研究中，我们确定了哪种炎性小体成分受 P2YR 激活调节，并参与乳腺癌细胞和放射抵抗 (RT-R)-乳腺癌细胞的肿瘤进展。首先，我们发现 NOD、LRR 和吡喃结构域蛋白 3 (NLRP3)；含 N 端 caspase 募集结构域 (CARD) 的 NLR 家族蛋白 4 (NLRC4)；凋亡相关斑点样蛋白含有 CARD 结构域 (ASC)；和 caspase-1mRNA 水平在 RT-R-MDA-MB-231 细胞中上调，而 TNF-α 或 ATP 处理诱导 NLRC4、ASC 和 caspase-1，但不诱导 NLRP3 蛋白水平。此外，TNF-α 或 ATP 以 P2YR 依赖的方式增加 MDA-MB-231 和 RT-R-MDA-MB-231 细胞中 NLRC4、ASC 和 caspase-1 的蛋白水平。此外，ATP 通过 P2YR 激活诱导 MDA-MB-231 和 RT-R-MDA-MB-231 细胞中 IL-1β 和 VEGF-A 的分泌以及侵袭，这被 NLRC4、ASC 和 caspase-1 小干扰 RNA (siRNA) 抑制。总之，本报告表明，ATP 通过 P2YR 激活诱导肿瘤侵袭和血管生成通过炎性小体激活，特别是通过调节炎性小体成分 NLRC4、ASC 和 caspase-1。
