The School of Pharmacy, University of London, 29-39 Brunswick Square, London WC1N 1AX, UK.
School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Liverpool L3 3AF, UK.
Int J Mol Sci. 2021 Jun 17;22(12):6498. doi: 10.3390/ijms22126498.
The constitutive expression or overactivation of cyclooxygenase (COX) and lipoxygenase (LOX) enzymes results in aberrant metabolism of arachidonic acid and poor prognosis in melanoma. Our aim is to compare the in vitro effects of selective COX-1 (acetylsalicylic acid), COX-2 (meloxicam), 5-LOX (MK-886 and AA-861), 12-LOX (baicalein) and 15-LOX (PD-146176) inhibition in terms of proliferation (SRB assay), mitochondrial viability (MTT assay), caspase 3-7 activity (chemiluminescent assay), 2D antimigratory (scratch assay) and synthesis of eicosanoids (EIA) in the B16F10 cell line (single treatments). We also explore their combinatorial pharmacological space with dacarbazine and temozolomide (median effect method). Overall, our results with single treatments show a superior cytotoxic efficacy of selective LOX inhibitors over selective COX inhibitors against B16F10 cells. PD-146176 caused the strongest antiproliferation effect which was accompanied by cell cycle arrest in G phase and an >50-fold increase in caspases 3/7 activity. When the selected inhibitors are combined with the antineoplastic drugs, only meloxicam provides clear synergy, with LOX inhibitors mostly antagonizing. These apparent contradictions between single and combination treatments, together with some paradoxical effects observed in the biosynthesis of eicosanoids after FLAP inhibition in short term incubations, warrant further mechanistical in vitro and in vivo scrutiny.
环氧化酶 (COX) 和脂氧合酶 (LOX) 酶的组成型表达或过度激活导致花生四烯酸代谢异常，黑色素瘤预后不良。我们的目的是比较选择性 COX-1（乙酰水杨酸）、COX-2（美洛昔康）、5-LOX（MK-886 和 AA-861）、12-LOX（黄芩素）和 15-LOX（PD-146176）在体外对 B16F10 细胞系增殖（SRB 测定）、线粒体活力（MTT 测定）、半胱天冬酶 3-7 活性（化学发光测定）、二维抗迁移性（划痕测定）和合成类二十烷酸（EIA）的抑制作用的差异（单药治疗）。我们还探索了它们与达卡巴嗪和替莫唑胺的组合药理学空间（中值效应法）。总体而言，我们的单药治疗结果表明，选择性 LOX 抑制剂对 B16F10 细胞的细胞毒性作用优于选择性 COX 抑制剂。PD-146176 引起的增殖抑制作用最强，伴随着细胞周期停滞在 G1 期，半胱天冬酶 3/7 活性增加 50 多倍。当选择的抑制剂与抗肿瘤药物联合使用时，只有美洛昔康提供明显的协同作用，而 LOX 抑制剂大多起拮抗作用。这些单药和联合治疗之间的明显矛盾，以及在短期孵育中 FLAP 抑制后观察到的类二十烷酸生物合成中的一些矛盾效应，需要进一步进行体外和体内机制研究。
