基于代谢组学与转录组学的结直肠癌湿热瘀毒证的代谢特征和分子机制研究  

Study on metabolic characteristics and molecular mechanism of colorectal cancer with damp-heat stasis toxin syndrome based on metabolomics and transcriptomics

作　　者：张烨 范旻旻 程海波[2,3] ZHANG Ye;FAN Minmin;CHENG Haibo(Department of Oncology,Jiangsu Province Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;The First Clinical College of Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine in Prevention and Treatment of Tumor,Nanjing 210023,China)

机构地区：[1]南京中医药大学附属医院,江苏省中医院肿瘤内科,南京210029 [2]南京中医药大学第一临床医学院,南京210023 [3]江苏省中医药防治肿瘤协同创新中心,南京210023

出　　处：《中华中医药杂志》2024年第5期2147-2154,共8页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点研发计划“中医药现代化”重点专项(No.2022YFC3500200,No.2022YFC3500202);国家自然科学基金重点项目(No.81930117)。

摘　　要：目的:观察湿热瘀毒证结直肠癌患者和健康人代谢轮廓及转录水平的基因差异,探索其潜在的代谢特征及分子机制。方法:本研究共纳入190例湿热瘀毒证结直肠癌患者及50名健康人,采用气相色谱-质谱联用(GC-MS)非靶向代谢组学检测两组间的代谢差异,同时通过有参转录组学技术检测其中10例患者肿瘤组织和正常组织的转录基因差异,进一步联合分析探索结直肠癌湿热瘀毒证的发生发展机制。结果:代谢组学分析结果显示两组代谢轮廓分离趋势较好,共筛选出160种差异代谢物,其中包括氨基酸、脂肪酸及尿素等代谢产物(P<0.05)。对差异代谢物进行KEGG富集分析,结果显示精氨酸-脯氨酸代谢及精氨酸合成等代谢通路在两组间具有显著性差异(P<0.05)。转录组测序结果显示,显著上调的差异基因有1 861个,下调的差异基因有1 271个。差异基因KEGG通路富集分析结果显示精氨酸-脯氨酸代谢通路为显著富集代谢通路(P<0.05)。综合代谢组学和转录组学分析结果,精氨酸-脯氨酸代谢通路为其共同代谢通路,荧光定量PCR结果显示:精氨酸转运蛋白SLC6A14、SLC7A1及该通路中代谢酶NOS1、ODC1在湿热瘀毒证肠癌组织中显著上调(P<0.05),而代谢酶OTC则显著下调(P<0.01)。以上趋势与转录组测序结果一致。为进一步分析这几个基因在湿热瘀毒证肠癌中的诊断价值,分别计算其受试者工作曲线下面积(AUC)的值,结果显示,AUC值均>0.80,具有较高的诊断价值。结论:精氨酸-脯氨酸代谢通路的过度激活可能与结直肠癌湿热瘀毒证的发生发展密切相关,该代谢通路中的关键基因靶点可作为结直肠癌湿热瘀毒证微观辨证的潜在生物标志物。Objective:To investigate the metabolic profile and transcriptomic gene differences between colorectal cancer patients with damp-heat stasis toxin syndrome and healthy subjects.Methods:A total of 190 colorectal cancer patients with dampheat stasis toxin syndrome and 50 healthy people were enrolled in the study.The metabolic differences between the two groups were detected using GC-MS with non-targeted metabolism,and the transcriptional gene differences between tumor tissue and normal tissue were detected using transcriptional techniques in 10 patients.A further comprehensive analysis was conducted to explore the mechanisms of occurrence and development of colorectal cancer with damp-heat stasis toxin syndrome.Results:The results of the metabolomics analysis showed a good trend in the separation of metabolic profiles between the two groups.A total of 160 different metabolites were screened,including amino acids,fatty acids and urea(P<0.05).KEGG enrichment analysis of differential metabolites showed that the metabolic pathways of arginine-proline metabolism and arginine synthesis were significantly activated between the two groups(P<0.05).Transcriptome sequencing results showed that there are 1861significantly up-regulated differential genes and 1271 significantly down-regulated differential genes.The KEGG pathway enrichment analysis indicated that the arginine-proline metabolic pathway is significantly enriched(P<0.05).The results of comprehensive metabolic and transcriptomic analyses indicated that the arginine-proline metabolic pathway is the same metabolic pathway.The results of fluorescence quantitative PCR showed that the arginine transporters SLC6A14,SLC7A1 and the metabolic enzymes NOS1 and ODC1 in this pathway were significantly up-regulated in the intestinal cancer tissues of patients with dampheat stasis toxin syndrome(P<0.05),while the metabolic enzyme OTC was significantly down-regulated(P<0.01).The above results were consistent with those from transcriptome sequencing.To further analyze the diagnostic

关 键 词：结直肠癌 湿热瘀毒证 代谢组学 转录组学 精氨酸-脯氨酸代谢 生物标志物 

分 类 号：R273[医药卫生—中西医结合]