哺乳动物雷帕霉素靶蛋白通路调控ECA109细胞放疗敏感性的代谢组学

doi:10.6040/j.issn.1671-7554.0.2019.940

摘要/Abstract

摘要： 目的探讨哺乳动物雷帕霉素靶蛋白(mTOR)通路对放疗诱导自噬的影响,寻找与放疗敏感性相关的小分子代谢物质及其调控通路。方法食管癌ECA109细胞分为对照组、放疗组和放疗联合MHY1485组;采用Western blotting法分别检测各组细胞mTOR、自噬相关蛋白Beclin-1和微管相关蛋白轻链3-II/I比值(LC3-II/I),采用CCK8法分别检测各组细胞经处理后24 h活性。收集放疗组和放疗联合MHY1485组细胞培养液,每组6例,采用液相色谱-质谱(LC-MS)技术对样本进行代谢组学检测,采用偏最小二乘判别分析(PLS-DA)和正交-偏最小二乘判别分析(OPLS-DA)法分析比较两组代谢物质差异。结果放疗可以通过抑制mTOR通路来诱导自噬的发生,MHY1485可以激活mTOR从而拮抗放疗诱导的自噬,抑制肿瘤的增殖。与放疗组相比,放疗联合MHY1485组细胞培养液中甜菜碱醛、肌酸、硬脂酸、鸟氨酸、L-胱氨酸和L-脯氨酸上调(P<0.001),瓜氨酸、烟酸、葡萄糖6-磷酸、L-缬氨酸、胸腺嘧啶、甜菜碱、L-精氨酸、L-亮氨酸、L-色氨酸和吡哆醇下调(P<0.001)。结论放疗可以抑制mTOR诱导的自噬,激活mTOR可以抵抗放疗诱导的自噬,从而抑制ECA109细胞增殖,增强放疗敏感性。食管癌ECA109细胞放疗组与放疗联合MHY1485组代谢产物不同,采用LC-MS技术检测可以区分放疗组与放疗联合MHY1485组,并发现与放疗敏感相关的代谢物质及其通路。

关键词: 哺乳动物雷帕霉素靶蛋白信号通路, 食管癌, 放射敏感性, 代谢组学

Abstract: Objective To explore the effects of mammalian target of rapamycin(mTOR)pathway on radiotherapy-induced autophagy, to search for the small molecular metabolites related to radiosensitivity and their regulatory pathways. Methods ECA109 cells were divided into three groups: control group, radiotherapy group and radiotherapy plus MHY1485 group. The mTOR, Beclin-1 and LC3-II/I were detected with Western blotting, and cell viability 24h after treatment was determined with CCK8 method. The cell culture medium of radiotherapy group(n=6)and radiotherapy plus MHY1485 group(n=6)were collected and analyzed with liquid chromatography-mass spectrometry(LC-MS). The differences in metabolites between the two groups were analyzed with PLS-DA and OPLS-DA. Results Radiotherapy induced autophagy by inhibiting mTOR pathway, and MHY1485 activated mTOR to antagonize autophagy and inhibited proliferation of tumor cells. Compared with the radiotherapy group, the radiotherapy plus MHY1485 group had upregulated expressions of betaine aldehyde, creatine, stearic acid, ornithine, L-cystine and L-proline(P<0.001), but downregulated expressions of citrulline, niacin, glucose 6-phosphate, L-valine, thymine, betaine, L-arginine, L-leucine, L-tryptophan and pyridoxine(P<0.001). Conclusion Radiotherapy can inhibit mTOR-induced autophagy, which 山东大学学报 (医学版)58卷1期 -章海容,等. 哺乳动物雷帕霉素靶蛋白通路调控ECA109细胞放疗敏感性的代谢组学 \=-can be resisted by the activation of mTOR, so as to inhibit the proliferation of ECA109 cells and enhance the radiosensitivity. As metabolites in the radiotherapy group are different from those in radiotherapy plus MHY1485 group, metabolites and their pathways related to radiosensitivity can be detected with LC-MS technology.

Key words: Mammalian target of rapamycin, Esophageal cancer, Radiosensitivity, Metabonomics

中图分类号:

R310.31

章海容, 张小红, 王超群. 哺乳动物雷帕霉素靶蛋白通路调控ECA109细胞放疗敏感性的代谢组学[J]. 山东大学学报 (医学版), 2020, 58(1): 6-12.

ZHANG Hairong, ZHANG Xiaohong, WANG Chaoqun. A metabonomic study on mTOR pathway regulating radiosensitivity of ECA109 cells[J]. Journal of Shandong University (Health Sciences), 2020, 58(1): 6-12.

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