基于核磁共振氢谱的胰腺癌组织代谢组学分析

Proton nuclear magnetic resonance spectroscopy-based metabonomic research on pancreatic cancer tissue

目的 观察胰腺癌组织、癌旁组织和正常组织代谢组的差异,进而筛选与胰腺癌发病相关的潜在生物标志物.方法 收集胰腺癌癌症患者手术切除下的胰腺导管腺癌组织（HPC） 19例,癌旁组织（HPP） 13例,正常组织（HPN）7例.采用甲醇-氯仿法萃取、上清液冻干等预处理后,采用核磁共振波谱并结合多变量统计分析处理,筛选出特征性代谢生物标志物和相应的代谢路径.结果 HPC-HPP和HPN-HPP建模效果不够理想;HPC-HPN建模成功,模型的解释率R2Y=0.765,预测率Q2=0.607,共筛选出24种差异代谢物.癌组织中1-甲基组氨酸、β-羟基丁酸、丙氨酸、一磷酸腺苷、三磷酸腺苷、乙醇胺、富马酸、甘氨酸、甘油磷脂酰胆碱、肌苷、异丁酸、甲硫氨酸、N-乙酰天冬氨酸、烟酰胺腺嘌呤二核苷酸、N-乙酰血清素、苯丙氨酸、丙酮酸、色氨酸、酪氨酸、尿嘧啶核苷、尿苷二磷酸葡萄糖的含量相对正常组降低,而亮氨酸和苏氨酸的含量在癌症组中升高.结论 成功建立了腺癌组织和正常组织模式识别,从中找到了可以用来区别癌症和正常组织的生物标志物.

Objective By studying the metabolic profile differences of pancreatic cancer tissue, paracancerous tissue and normal pancreatic tissue, find out some significant biomarkers for the pancreatic cancer diagnosis and further researches.Methods 19 cases of pancreatic ductal adenocarcinoma （HPC）, 13 cases of paracancerous tissues （HPP） and 7 cases of normal pancreatic tissue（HPN） were collected from the pancreatic cancer patients.The pancreatic tissues from each group underwent pre-treatments including methanol-chloroform extraction and freeze drying of supernate.NMR spectroscopy was applied to screen out the potential metabolic biomarkers and the corresponding pathways relevant to normal pancreatic tissue and pancreatic cancer tissue by combination with the multivariate statistical analyses.Results The modelings of paracancerous tissue vs cancer tissue （HPC-HPP）, paracancerous tissue vs normal tissue （HPN-HPP） are invalid, however carcinoma tissue vs normal tissue（HPC-HPN） modeling is successfully constructed and stable, R2Y of the model is 0.765, Q2 of the model is 0.607.The metabolic analysis revealed relatively increased level of leucine, threonine, and lower level of 1-methylhistdine, 3-hydroxybutyrate, alanine, adenosine phosphate （AMP）, adenosine triphosphate （ATP）, ethanolamine, fumarate, glycine, glycerol phosphatidylcholine （GPC）, inosine, isobutyrate, methoinine, N acetyl aspartic acid （NAA）, nicotinamide adenine dinucleotide （NAD）, N-acetyl serotonin （NAS）, phenylalanine （Phe）, pyruvate, tryptophan, tyrosine, uridine, uridine diphosphate glucose two phosphate （UDPG） in the pancreatic cancer tissue compared to that of normal pancreatic tissue.Conclusion The metabolic profile of pancreatic cancer tissue and normal pancreatic tissue is significantly discriminated and the biomarkers may contribute to find the therapeutic target of pancreatic cancer and its further research.