我们旨在研究间充质干细胞分泌物和甲氨蝶呤的组合对神经母细胞瘤细胞的5-羟色胺2A（5-HT2AR）、5-羟色胺7（5-HT7R）、腺苷2A（A2AR）受体以及CD73等生物特征的影响。甲氨蝶呤作为神经母细胞瘤细胞的血清素拮抗剂。我们使用人牙髓来源的干细胞（hDPSCs）来获取条件培养基（CM）。将甲氨蝶呤药物制备在CM中并应用于神经母细胞瘤细胞。通过Western blot和免疫荧光染色分析5-HT7R、5-HT2AR、A2AR和CD73的表达。根据产品程序使用生物活性测试试剂盒进行总的细胞凋亡、线粒体膜去极化、Ki-67增殖试验、细胞活力分析、DNA损伤和细胞周期分析。我们的结果表明，神经母细胞瘤癌细胞通常位于血清素7受体和腺苷2A受体的Gs信号轴上。CM和甲氨蝶呤抑制了神经母细胞瘤细胞中的5-HT7和A2A受体水平。我们发现，CM和甲氨蝶呤形成了5-HT2AR、5-HT7R、A2AR和CD73之间的交叉抑制作用。CM和甲氨蝶呤增加了神经母细胞瘤细胞的总凋亡率，并诱导了线粒体膜去极化。CM和甲氨蝶呤诱导了DNA损伤，并使神经母细胞瘤细胞在G0/G1期阻滞。这些结果表明，CM和甲氨蝶呤的组合可能对神经母细胞瘤癌细胞产生治疗效果，未来的体内研究对支持这些发现在神经母细胞瘤研究领域中可能是重要的。版权所有©2023 Elsevier B.V. 发表。

(s): We aimed to investigate the effects of mesenchymal stem cell secretome and methysergide combination on 5-hydroxytryptamine 2A, (5-HT2AR), 5-hydroxytryptamine 7 (5-HT7R), adenosine 2A (A2AR) receptors and CD73 on neuroblastoma cell line and how they affect biological characteristics. Methysergide was used as a serotonin antagonist on the neuroblastoma cells.Human dental pulp-derived stem cells (hDPSCs) used to obtain conditioned medium (CM). Methysergide drug was prepared in CM and applied to neuroblastoma cells. Analysis of 5-HT7R, 5-HT2AR, A2AR and CD73 expressions was performed by western blot and immunofluorescence staining. Total apoptosis, mitochondrial membrane depolarization, Ki-67 proliferation test, viability analysis, DNA damage and cell cycle analysis were performed in accordance with the product procedure by using biological activity test kits.Our results showed that neuroblastoma cancer cells are normally on the Gs signaling axis via the serotonin 7 receptor and the adenosine 2A receptor. CM and Methysergide inhibited the 5-HT7 and A2A receptor levels in neuroblastoma cells. We found that CM and methysergide formed crosstalk inhibition between 5-HT2AR, 5-HT7R, A2AR and CD73. CM and Methysergide increased the total apoptosis in neuroblastoma cells and induced the mitochondrial membrane depolarization. CM and Methysergide induced the DNA damage and arrested in G0/G1 phase of cell cycle of the neuroblastoma cells.These findings suggest that the combination of CM and methysergite may exert a therapeutic effect on neuroblastoma cancer cells, and future in vivo studies may be important in area of neuroblastoma research to support the findings.Copyright © 2023. Published by Elsevier B.V.