Tumor associated macrophages (TAMs) have a crucial role in cancer progression, metastasis and drug response. Piroxicam and sulindac sulfide are non-steroidal anti-inflammatory drugs (NSAID) that decrease the incidence and progression of several types of cancer. However, their role in suppressing the interactions between TAMs and cancer cells remain unclear. Herein, we studied the impact of human monocytes conditioned media (CM) on cellular proliferation of ER-dependent MCF-7 and ER-independent MDA-MB-231 cells, and the effects of piroxicam and sulindac sulfide on the expression levels of RAS, COX-2, IL-6, IL-1β and PAR-4 (qRT-PCR), BCL-2 and BAX (western blot), Caspase-3, VEGF-a and PGE2 (ELISA), MMP-2 and -9 (zymography) in the stimulated cells. Our results showed that CM caused a significant increase in cells survival through significant increase in RAS expression which resulted in upregulation of COX-2, PGE2, BCL-2, IL-6, IL-1β, VEGF-A and MMP-9 and down regulation of PAR-4. Treatment with one of the NSAIDs used in this study produced a time and concentration dependent growth inhibition of stimulated cells by inhibiting RAS expression. Suppression of RAS was accompanied by downregulation of its downstream signaling of IL-1β, IL-6, COX-2 and PGE2, activation of apoptotic machinery through upregulation of PAR-4 and caspase-3, as well as, inhibition of BCL-2, VEGF-A, MMP-2 and MMP-9. In conclusion, our data support the role of piroxicam and sulindac sulfide in suppressing inflammation-driven breast cancer progression and identifies promising novel target in RAS and PAR-4 signaling.

中文翻译：

---

巨噬细胞的抑制-通过靶向乳腺癌细胞系中的RAS和PAR-4信号传导诱导炎症。

肿瘤相关巨噬细胞（TAM）在癌症进展，转移和药物反应中起着至关重要的作用。吡罗昔康和舒林酸硫化物是非甾体类抗炎药（NSAID），可降低几种癌症的发生率和进展。然而，它们在抑制TAM与癌细胞之间相互作用中的作用仍不清楚。在这里，我们研究了人类单核细胞条件培养基（CM）对ER依赖性MCF-7和ER依赖性MDA-MB-231细胞增殖的影响，以及吡罗昔康和硫化舒林酸对RAS表达水平的影响， COX-2，IL-6，IL-1β和PAR-4（qRT-PCR），BCL-2和BAX（蛋白质印迹），Caspase-3，VEGF-a和PGE2（ELISA），MMP-2和-9 （酶法）在刺激的细胞中。我们的结果表明，CM通过显着增加RAS表达而导致细胞存活率显着增加，从而导致COX-2，PGE2，BCL-2，IL-6，IL-1β，VEGF-A和MMP-9上调和下调PAR-4的规定。本研究中使用的一种NSAIDs的治疗通过抑制RAS表达，对受刺激的细胞产生了时间和浓度依赖性的生长抑制。RAS抑制伴随其IL-1β，IL-6，COX-2和PGE2下游信号的下调，通过PAR-4和caspase-3的上调激活凋亡机制以及抑制BCL-2 ，VEGF-A，MMP-2和MMP-9。总之，我们的数据支持吡罗昔康和硫化舒林酸在抑制炎症驱动的乳腺癌进展中的作用，并在RAS和PAR-4信号传导中鉴定出有希望的新靶标。