Shanxi Medical University School of Stomatology
【摘要】目的 通过检测口腔癌皮下移植瘤小鼠小肠中代谢物和代谢通路的变化，分析过表达miR-181a-5p对皮下移植瘤小鼠小肠中代谢组和代谢物组的影响。方法 实验分为三组，空白对照组(Control)，阴性对照组(Negative control,NC组)以及实验组(Over expression of miR-181a-5p,OE组)。将不同组别处理后的细胞混悬液，通过皮下注射到M-NSG重度免疫缺陷雌性小鼠右侧腹股沟中上部，构建成口腔癌皮下移植瘤小鼠模型。按时记录小鼠体重变化，对小鼠小肠组织进行HE染色，观察各组病理变化。采用超高效液相色谱-串联飞行时间质谱联用仪和串联Orbitrap质谱联用仪检测NC 组、OE组和 Control组小鼠小肠中的代谢物，使用XCMS预分析原始数据，质量评价样本数据，鉴定Control组与NC组、NC组与OE组的差异代谢物，进行KEGG富集分析获取差异代谢通路。结果 Control和 NC组小肠组织中共鉴定出170种差异代谢物。代谢物富集的显著性信号通路包括胆碱代谢，丙氨酸、天冬氨酸和谷氨酸代谢，γ-氨基丁酸(GABA)神经突触代谢，甘油磷脂代谢，环磷酸腺苷(cAMP)信号通路代谢，癌症中心碳代谢以及烟酸和烟碱胺代谢通路。NC组和OE组相比，小鼠小肠中检测到VIP(Variable Importance in the Projection)值大于2的差异代谢物有16种，显著性差异代谢物包括甘油磷酰胆碱、棕榈酸、3-羟基丁酰肉碱、β-羟丁酸等。代谢物富集到的显著性差异通路为胆碱代谢通路。 结论 口腔癌皮下移植瘤可引起小鼠小肠的代谢物发生变化，主要改变了小肠中与能量代谢相关的代谢物。过表达miR-181a-5p影响口腔癌皮下移植瘤小鼠小肠代谢物，代谢物富集通路为胆碱代谢通路。
[Abstarct] Objective By detecting changes in metabolites and metabolic pathways in the small intestine of mice with subcutaneous transplantation of oral cancer, the effects of overexpression of miR-181a-5p on metabolites and metabolites in the small intestine of mice with subcutaneous transplantation of oral cancer were analyzed. Methodes Three groups existed in the experiment: the Control group, the Negative control group, and the Over expression of miR-181a-5p group in the experimental group. To construct a subcutaneous transplantation tumor mouse model of oral cancer, different groups of treated cell suspensions were subcutaneously injected into the right point and upper location of the groin of M-NSG severely immunodeficient female mice. The pathogenic changes in each group were identified while additionally following the changes in the mice's body weight and small intestinal tissues using HE staining. By using tandem Orbitrap mass spectrometry and ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry, the metabolites in the small intestine of mice in the NC group, OE group, and Control group have been detected. By pre-analyzing the original data and quality rating sample data, XCMS was able to assess which metabolites were different between the Control group and NC group and between NC group and OE group. To establish the unique metabolic pathways, KEGG enrichment analysis was used. Results A total of 170 distinct metabolites were found in the small intestinal tissues of the Control and NC groups. Choline metabolism, alanine, aspartate, and glutamate metabolism, GABA synaptic metabolism, glycerophospholipid metabolism, cAMP signaling route, cancer center carbon metabolism, and niacin and niacin amine metabolic pathways are important signaling pathways for metabolite enrichment. In the NC group, 16 distinct metabolites with VIP values larger than 2 were found in the small intestine of mice compared to the OE group that overexpressed miR-181a-5p. Glycerin phosphoylcholine, palmitic acid, 3-hydroxybutyryl carnitine, -hydroxybutyric acid, etc. are example of the metabolites which significantly vary. The primary raised metabolism path is the one for choline. Conclusions Mice's small intestine suffered slight changes as a result of subcutaneous transplantation of oral cancer, with the greatest effect in the metabolites critical in energy metabolism. The choline metabolic path was the pathway that selected absolutely metabolites in the small intestine of mice with the subcutaneous grafts of oral cancer.