Nguyen Phu Hung, Touchefeu Yann, Durand Tony, Aubert Philippe, Duchalais Emilie, Bruley des Varannes Stanislas, Varon Christine, Neunlist Michel, Matysiak-Budnik Tamara
1 Université Bretagne Loire, Université de Nantes, INSERMU1235, TENS, Institut des Maladies de l'Appareil Digestif du CHU, Nantes, France.
2 Faculty of Biotechnology, Thai Nguyen University of Sciences, Thai Nguyen, Vietnam.
Tumour Biol. 2018 Sep;40(9):1010428318799028. doi: 10.1177/1010428318799028.
Gastric cancer is the third leading cause of cancer-related death worldwide, but the mechanisms of gastric carcinogenesis are not completely understood. Recently, the role of cholinergic neuronal pathways in promoting this process has been demonstrated. Our aim was to extend these studies and to evaluate, using an in vitro model of tumorspheres, the effect of acetylcholine on human gastric cancer cells, and the role of acetylcholine receptors and of the nitric oxide pathway, in this effect. The gastric cancer cell line MKN-45 of the diffuse type of gastric cancer was cultured in the presence of acetylcholine, or different agonists or inhibitors of muscarinic and nicotinic acetylcholine receptors, or nitric oxide donor or inhibitor of the nitric oxide pathway, and the number and size of tumorspheres were assessed. The expression of cancer stem cell markers (CD44 and aldehyde dehydrogenase) was also evaluated by immunofluorescence and quantitative reverse transcription polymerase chain reaction. We showed that acetylcholine increased both the number and size of tumorspheres and that this effect was reproduced with both muscarinic and nicotinic acetylcholine receptors agonists and was inhibited by both receptor antagonists. The nitric oxide donor stimulated the tumorsphere formation, while the nitric oxide synthesis inhibitor inhibited the stimulatory effect of acetylcholine. Moreover, acetylcholine increased the expression of stem cell markers on gastric cancer cells. These results indicate that acetylcholine induces the stem cell properties of gastric cancer cells and both muscarinic and nicotinic receptors and a nitrergic pathway might be involved in this effect.
胃癌是全球癌症相关死亡的第三大主要原因，但胃癌发生的机制尚未完全明确。最近，胆碱能神经元通路在促进这一过程中的作用已得到证实。我们的目的是扩展这些研究，并使用肿瘤球体外模型评估乙酰胆碱对人胃癌细胞的影响，以及乙酰胆碱受体和一氧化氮途径在这一影响中的作用。将弥漫型胃癌的胃癌细胞系MKN - 45在乙酰胆碱、毒蕈碱型和烟碱型乙酰胆碱受体的不同激动剂或抑制剂、一氧化氮供体或一氧化氮途径抑制剂存在的情况下进行培养，评估肿瘤球的数量和大小。还通过免疫荧光和定量逆转录聚合酶链反应评估癌症干细胞标志物（CD44和醛脱氢酶）的表达。我们发现乙酰胆碱增加了肿瘤球的数量和大小，毒蕈碱型和烟碱型乙酰胆碱受体激动剂均可重现这一效应，且均被受体拮抗剂抑制。一氧化氮供体刺激肿瘤球形成，而一氧化氮合成抑制剂抑制乙酰胆碱的刺激作用。此外，乙酰胆碱增加了胃癌细胞上干细胞标志物的表达。这些结果表明，乙酰胆碱诱导胃癌细胞的干细胞特性，毒蕈碱型和烟碱型受体以及一氧化氮能途径可能参与了这一效应。
