基于LC-MS技术的患乳腺癌猫血清代谢组学分析

Serum Metabolomics Analysis of Feline Mammary Carcinomas Based on LC-MS Techniques

本试验旨在研究患乳腺癌猫与健康猫体内差异代谢物的变化情况,并探讨差异代谢物与猫乳腺癌发生之间的关系。本试验选取临床收集的经组织病理学确诊的猫乳腺癌血清样本6例作为试验组(T组),同时选取年龄相似,品种相同的健康猫血清6例作为对照组(C组)。运用超高效液相色谱质谱联用技术(liquid chromatography-mass spectrometry,LC-MS)对两组猫的血清样本进行检测,采用无监督的主成分分析(principal component analysis,PCA)、正交偏最小二乘法-判别分析(orthogonal projections to latent structures-discriminant analysis,OPLS-DA)及学生t检验(Student's t-test)筛选出差异代谢物,然后再对筛选出的差异代谢物进行层次聚类分析(hierarchical clustering analysis,HCA),并进行差异代谢物的KEGG注释及差异代谢物的代谢通路分析。结果表明,在两组样本中共定性到159种差异代谢物。与C组相比,在T组中有49种差异代谢物出现下调,110种差异代谢物出现上调。最终共筛选出5种与猫乳腺癌发生发展密切相关的差异代谢产物。与C组相比,T组中麦角硫因(ergothioneine,EGT)和肌酸(creatine)出现下调,吲哚乙酸(indolelactic acid,IAA)、胆碱(choline)和尿酸(uric acid)出现上调。这些差异代谢物表明,在猫乳腺癌的发生过程中,机体变化涉及了甘氨酸、丝氨酸和苏氨酸代谢、精氨酸和脯氨酸代谢、组氨酸代谢、色氨酸代谢、嘌呤代谢异常和甘油磷脂代谢等多个代谢途径,为今后深入研究猫乳腺癌的发病机制开辟了一个新的思路。

The purpose of this study was to investigate the variation of differential metabolites in mammary carcinomas bearing cats and healthy cats,and to explore the relationship between differential metabolites and the occurrence of feline mammary carcinomas.In this study,6 feline mammary carcinomas serum samples were selected as test(T)group.Meanwhile,6 healthy cats with similar age and same breed were selected as control(C)group.Serum samples were detected by ultra-high performance liquid tandem chromatography quadrupole time of flight mass spectrometry(LC-MS).Differential metabolites were screened by principal component analysis(PCA),orthogonal projections to latent structures-discriminant analysis(OPLS-DA)and Student's t-test.Then the hierarchical clustering analysis(HCA)was carried out for the screened differential metabolites.The KEGG annotation and the metabolic pathway of differential metabolites were analyzed.The results showed that a total of 159 differential metabolites were identified in the 2 groups.Compared with C group,49 differential metabolites were down-regulated and 110 differential metabolites were up-regulated in T group.Finally,a total of 5 differential metabolites which closely related to feline mammary carcinomas were selected.Ergothioneine(EGT)and creatine in T group were down-regulated,while indolelactic acid(IAA),choline and uric acid were up-regulated compared to C group.These differential metabolites indicated that during the development of feline mammary carcinomas,the body changes involved multiple metabolic pathways,such as glycine,serine and threonine metabolism,arginine and proline metabolism,histidine metabolism,tryptophan metabolism,purine metabolism and glycerophospholipid metabolism.This study provides a new idea for further research of the pathogenesis of feline mammary carcinomas.