mTORC1抑制剂影响谷氨酰胺代谢抑制人胰腺神经内分泌肿瘤细胞增殖

摘要

Objective To evaluatethe effect of mTORC1 inhibitor on the proliferation in human pancreatic neuroendocrine tumors(pNET)cell line BON,to explore the function of glutamine metabolism in it.Methods In vitro cultured human pancreatic neuroendocrine tumors(pNET)cell line BON,BON cells were treated with different concentrations of rapamycin(1,5,10,25,50, 100 nM)for 12,24 h.Then CCK-8 assay are used to calculate the growth inhibitory rate.Rapamycin treated with BON 12 h,test the glutamine uptake level compared with control.Then deprive of glucose and/or glutamine,CCK-8 assay were used in observation of cell proliferation,cell cycle distribution was analyzed by flow cytomety.Results Rapamycin significantly inhibited the growth of BON cells in a time-and dose-dependent manner(P <0.05).Meanwhile,rapamycin can reduce the glutamine uptake level compared with control.BON obviously depends on glutamine for growth,without glucose and glutamine group have obvious difference in growth rate(P <0.05).Conclusion mTORC1 inhibitor can inhibit BON cells proliferation and influence the glutamine uptake lev-el.suggesting that mTORC1 inhibitor might inhibit BON cells proliferation by influenced the glutamine metabolic pathway.%目的：体外研究雷帕霉素靶蛋白复合物1（mTORC1）抑制剂对人胰腺神经内分泌肿瘤（pNET）细胞株 BON 增殖的影响，探讨谷氨酰胺（Gln）代谢在 mTORC1抑制剂影响 BON 细胞增殖中的作用。方法体外培养人胰腺神经内分泌肿瘤细胞BON，分别用1、5、10、25、50、100 nM 的雷帕霉素处理后，应用 CCK-8检测 BON 的生长抑制率。用100 nM 雷帕霉素处理 BON细胞12 h 后，检测其对 Gln 的吸收水平。在无葡萄糖（Glc）和（或）无谷氨酰胺（Gln）的情况下，应用 CCK-8检测 BON 的增殖情况以及流式细胞术检测细胞周期情况。结果雷帕霉素可明显抑制 BON 细胞的增殖，抑制率呈时间-浓度依赖性（P ＜0．05）。同时雷帕霉素影响 BON 对 Gln 的吸收，而 BON 明显依赖于 Gln 生长，Glc－／Gln＋组、Glc－／Gln－组、Glc＋／Gln－组生长速率较 Glc＋／Gln＋组差异有统计学意义（P ＜0．05）。结论 mTORC1抑制剂可能通过影响 Gln 代谢抑制 BON 细胞的增殖。