| Background:Endometrial carcinoma seriously does harm to the psychological and physical health of female and locates the sixth most common gynecologic cancer throughout the world,affecting 319,600 women in 2015.With the development of economics in our country,the morbidity of endometrial carcinoma with hypertension,obesity,diabetes or hyperlipidaemia rapidly increased in recent years.Type I endometrial carcinomas are usually relates to unopposed estrogen stimulation,and is the most common subtype(80%).More and more endometrioid endometrial carcinomas occurred in the young femal patients in recent years,of which 5?10%under 40 years old.Hence,the early diagnosis and therapy supervising of type ? endometrial carcinomas are of great significance.As a kind of hormonal dependent tumor,type ? endometrial carcinomas possesses a few of abnormal metabolic pathways.Then metabolomics,depicting the metabolism rules of organisms,afford experimental basis for achieve this goal.On one hand,metabolomics could detect and quantize all the endogenous metabolites and their dynamic variation rules,contributing to look for the effective endogenous biomarkers(nuclease,amino acid,fatty acid and carbohydrate,etc)for the diagnosis and individualized monitoring.On the other hand,metabolomics could clarify the changing rules of diseases,and reveal the biological procedding principle of diseases to offer a new perspective of tumour research.Objective:Based on GC-TOF-MS and UHPLC-MS/MS metabolomic methods,we plan to filtrate the differential metabolites between type I endometrial carcinomas and disease control group patients in serum and urine.And then map the differential metabolites to the related databases and search for the differential metabolomic pathways.At last,we intend to verify the differential metabolites in endometrioid endometrial carcinomas tissue and para-carcinoma tissue using metabolomics target analysis,and explore the application of pathogenetic mechanism and auxiliary diagnosis and treatment.Methods:According to the access conditions,we selected 30 type I endometrial carcinomas patients and 31 disease control group patients.Serum and urine samples of every patients and the endometrioid endometrial carcinomas tissue and para-carcinoma tissue samples were all preserved in-80?.All the samples were detectd and identificated using non-target GC-TOF-MS after the derivatization by BSTFA(with 1%TMCS,v/v).The primary data were analyzed and modeled by PC A and OPLS-DA,followed by filtrating and mapping the differential metabolites in the KEGG database.After selecting the differential pathways,we verified the differential metabolites in endometrioid endometrial carcinomas tissue and para-carcinoma tissue using metabolomics target UHPLC-MS/MS analysis to confirm the differential metabolites and corresponding pathways.Results:In the serum GC-TOF-MS non-target metabolomics study,we obtained 354 peaks in all,reserved 274 peaks at last.After PCA and OPLS-DA analysis,29 differential metabolites were confirmed.Among them,the content of 9 differential metabolites,including alanine,iminodiacetic acid,uracil,d-glucoheptose,oxalacetic acid,N-acetyl-D-galactosamine,malonamide,N,N-dimethylarginine,N-acetyltryptophan increased obviously,and the content of 20 differential metabolites,including palmitic acid,3-hydroxybutyric acid,linoleic acid,palmitoleic acid,glutamine,heptadecanoic acid,taurine,2-ketobutyric acid,myristic acid,putrescine,trans-3,5-dimethoxy-4-hydroxycinnamaldehyde,N-acetyl-5-hydroxytryptamine,Bis(2-hydroxypropyl)amine,N-acetyl-L-leucine,5-hydroxyindole-2-carboxylic acid,5-hydroxytryptophan,N(epsilon)-trimethyllysine,tartaric acid,cyclohexylamine and L-cysteine decreased remarkblly.The metabolites pathway analysis displayed that the change of fatty acide biosynthesis,pantothenate and CoA biosynthesis,glyoxylate and dicarboxylate metabolism,citrate cycle,taurine and hypotaurine metabolism,linoleic acid metabolism,pyrimidine metabolism,arginine and proline metabolism played an essential role in type I endometrial carcinomas patients serum,according to the serum non-target metabolites analysis.In the urine GC-TOF-MS non-target metabolomics study,we obtained 477 peaks in all,and reserved 299 peaks at last.After PCA and OPLS-DA analysis,29 differential metabolites were confirmed.Among them,the content of 7 differential metabolites,including creatine,sucrose,phosphate,lactobionic acid,noradrenaline,DL-dihydrosphingosine,N-acetyl-beta-alanine increased obviously,and the content of 22 differential metabolites,including fucose,uracil,D-galacturonic acid,saccharic acid,3-aminoisobutyric acid,xanthine,L-cysteine,guanidinosuccinic acid,L-malic acid,2-hydroxyvaleric acid,fumaric acid,purine riboside,1,5-anhydroglucitol,sophorose,digalacturonic acid,inosine,1-kestose,oxamic acid,carbobenzyloxy-1-leucine degr4,3,5-dihydroxyphenylglycine,5,6-dimethylbenzimidazole,glutaconic acid decreased remarkblly.The metabolites pathway analysis indicated that the change of citrate cycle,pantothenate and CoA biosynthesis,beta-alanine metabolism,glycine serine and threonine metabolism,starch and sucrose metabolism,pyrimidine metabolism,purine metabolism,cysteine and methionine metabolism played an essential role in type I endometrial carcinomas patients urine,according to the urine non-target metabolites analysis.Combining serum and urine differential metabolites results using non-target GC-TOF-MS metabolomics analysis and the literature results of non-target metabolomics analysis in endometrioid endometrial carcinomas tissue,we selected 6 micromolecule compounds,namely palmitic acid,L-cysteine,uracil,taurine,inosine and linoleic acid,and further verified these compounds in endometrioid endometrial carcinomas tissue and para-carcinomas tissue,using target metabolomics analysis.As for the different physicochemical property,these 6 compounds were analyzed by GC-Q-MS and UHPLC-MS/MS respectively.The final experimental results indicated that the content of uracil,L-cysteine and taurine were different dramatically between the carcinoma tissue and para-carcinoma tissue(p<0.05),which afforded steady theory and experimental basis for the following metabolomics molecular pathways and function research.Conclusion:1.The serum and urine non-target GC-TOF-MS metabolomics analysis indicated that 56 differential metabolites were confirmed,and the final KEGG pathway analysis were performed;2.Using target GC-Q-MS and UHPLC-MS/MS metabolomics analysis,parts of filtrated differential metabolites from serum and urine were performed verification,and the results displayed that the content of uracil,L-cysteine and taurine were different dramatically between carcinoma and para-carcinoma tissue.Uracil,L-cysteine and taurine related metabolomics pathways had significant effect during the development of type I endometrioid endometrial carcinomas. |
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