Akimov Mikhail G, Gretskaya Natalia M, Gorbacheva Evgenia I, Khadour Nisreen, Sherstyanykh Galina D, Bezuglov Vladimir V
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia.
Moscow Center for Advanced Studies, Kulakova Str. 20, 123592 Moscow, Russia.
Int J Mol Sci. 2025 Jan 19;26(2):820. doi: 10.3390/ijms26020820.
2-arachnadoyl glycerol (2-AG) is one of the most common endocannabinoid molecules with anti-proliferative, cytotoxic, and pro-proliferative effects on different types of tumors. Typically, it induces cell death via cannabinoid receptor 1/2 (CB1/CB2)-linked ceramide production. In breast cancer, ceramide is counterbalanced by the sphingosine-1-phosphate, and thus the mechanisms of 2-AG influence on proliferation are poorly understood. We evaluated the mechanism of the anti-proliferative action by 2-AG and the influence of lysophaosphatidylinositol (LPI) on it in six human breast cancer cell lines of different tumor degree (MCF-10A, MCF-7, BT-474, BT-20, SK-BR-3, and MDA-MB-231) using resazurin test, inhibitor, blocker, and anti-oxidant analysis, and siRNA interference. To avoid acyl migration in 2-AG, we replaced it with the analog 2-arachidonoyl-1,3-difluoropropanol (2-ADFP) newly synthesized by us. Using a molecular docking approach, we showed that at the CB2 receptor, 2-ADFP and 2-AG were very close to each other. However, 2-ADFP demonstrated a stronger affinity towards CB1 in the antagonist-bound conformation. 2-ADFP was anti-proliferative in all the cell lines tested. The toxicity of 2-ADFP was enhanced by LPI. 2-ADFP activity was reduced or prevented by the CB2 and vanilloid receptor 1 (TRPV1) blockers, inositol triphosphate receptor, CREB, and cyclooxygenase 2 inhibitor, and by anti-oxidant addition. Together with the literature data, these results indicate CB2- and TRPV1-dependent COX-2 induction with concomitant cell death induction by the oxidized molecule's metabolites.
2-花生四烯酸甘油酯（2-AG）是最常见的内源性大麻素分子之一，对不同类型的肿瘤具有抗增殖、细胞毒性和促增殖作用。通常，它通过大麻素受体1/2（CB1/CB2）相关的神经酰胺生成诱导细胞死亡。在乳腺癌中，神经酰胺被1-磷酸鞘氨醇所平衡，因此2-AG对增殖影响的机制尚不清楚。我们使用刃天青试验、抑制剂、阻断剂和抗氧化剂分析以及小干扰RNA（siRNA）干扰，评估了2-AG的抗增殖作用机制以及溶血磷脂酰肌醇（LPI）对其在六种不同肿瘤程度的人乳腺癌细胞系（MCF-10A、MCF-7、BT-474、BT-20、SK-BR-3和MDA-MB-231）中的影响。为避免2-AG中的酰基迁移，我们用我们新合成的类似物2-花生四烯酰基-1,3-二氟丙醇（2-ADFP）替代了它。使用分子对接方法，我们表明在CB2受体处，2-ADFP和2-AG彼此非常接近。然而，2-ADFP在拮抗剂结合构象中对CB1表现出更强的亲和力。2-ADFP在所有测试的细胞系中均具有抗增殖作用。LPI增强了2-ADFP的毒性。CB2和香草酸受体1（TRPV1）阻断剂、肌醇三磷酸受体、CREB和环氧化酶2抑制剂以及添加抗氧化剂可降低或阻止2-ADFP的活性。结合文献数据，这些结果表明氧化分子的代谢产物通过CB2和TRPV1依赖性诱导环氧化酶2，同时诱导细胞死亡。
