Analysis Of Urinary Modified Nucleosides By LC-MS And Its Value In The Diagnosis And Treatment Of Breast Cancer With TCM And Western Medicine

Objective To establish a multi-component analysis method of urine modified nucleosides which could be used in clinical study,and to investigate the clinical value of urinary modified nucleosides in the diagnosis and treatment of breast cancer.Methods ①The procedure of the pretreatment and analysis of the sample were stand ardized and fulfilled by optimizing and improving the process.And a method of the multi-components detection of urinary modified nucleosides was established.②According to the inclusion/exclusion criteria,the patients and healthy people who were treated in Zhejiang Province Traditional Chinese Medical Hospital from September 2016 to July 2017 were divided into breast cancer group(55 cases),benign breast diseases group(55 cases)and healthy control group(54 cases).The urinary modified nucleosides were quantitatively analyzed via the assay platform.We investigate the relationship between urinary modified nucleosides and clinical features of breast cancer by the statistical analysis which was conducted on the data of participants’ information(age,menopausal state,BMI,tumor size,pathological type,lymph node metastasis,TNM stage,molecular subtypes,TCM syndrome differentiation and operation)and the comparison between modified nucleosides and CA153.Results ①6 modified nucleosides(except Pseu,NNdmG)calibrators showed a linear response from 1 to 800ng/mL(R2>0.9942),NNdmG calibrator showed a linear response from 20 to 3200ng/mL(R2=0.9996),The limitation of detection were no more than 1.0ng/mL;Pseu calibrator showd a linear response from 1 to 20μg/mL(R2=0.9972),The limitation of detection were no more than 10.0ng/mL.②Age,menopausal status and BMI did not affect the levels of 8 modified nucleosides(5mU,Pseu,N4C,NNdmG,N2mG,1mG,3mU,1mA)in healthy controls(P>0.05);The level of 7 modified nucleosides(5 mU,Pseu,N4C,NNdmG,N2mG,1mG,3mU)in breast cancer group were much hig her than those of benign breast diseases group and healthy control group(p<0.01),But there was no statistical difference of 1mA among three groups(P>0.05);The positive incidence of 7 modified nucleosides(5mU,Pseu,N4C,NNdmG,N2mG,1mG,3mU)were significantly higher than CA153(P<0.05);Logistic regression and ROC curve were aplied to analyze the date and evaluate the diagnostic values of each modified nucleosides,The regression equation of joint diagnostic predictive value was established:YBreast cancer =3.557*mG+2.979*N4C+2.043*NNdmG-12.269;The AUC of joint nucleosides(0.96)>The AUC of NNdmG(0.933)>The AUC of other modified nucleosides,The sensitivity of joint nucleosides was 96.4%and the specificity was 87.2%;There was no correlation of 7 modified nucleosides(5mU,Pseu,N4C,NNdnG,N2mG,1mG,3mU)to TNM stage and tumor size(P>0.05);Among the breast cancer patients with different pathological types,molecular subtypes and TCM syndrome differentiation,the levels of 7 modified nucleosides(5mU,Pseu,N4C,NNdmG,N2mG,1mG,3mU)were not statistically different(P>0.05);The level of 7 modified nucleosides(5mU,Pseu,N4C,NNdmG,N2mG,1mG,3mU)in breast cancer patients with node metastasis were significantly higher than the patients without node metastasis(P<0.05);Compared with the results of the day before operation,the levels of 7 modified nucleosides(5mU,Pseu,N4C,NNdmG,N2mG,1mG,3mU)were significantly decreased 7days after Radical Mastectomy and Modified Radical Mastectomy for breast cancer(P<0.05).Conclusion The constructed LC-MS method is suitable for the detection of urinary modified nucleosides;Urinary modified nucleosides may become novel non-invasive turn or markers with the high diagnostic efficiency and the analysis of modified nucleosides is ofgreat value in aiding early diagnosis,tumor distant metastasis,surveillance after primary surgey.