Dissertation
Dissertation > Medicine, health > Oncology > Genitourinary tumors > Breast tumor

Preliminary Metabolomics Study of Breast Cancer Based UPLC-Q/TOF and ~1 HNMR

Author FuFangFang
Tutor FengYiFan
School Guangdong College of Pharmacy
Course Drug analysis
Keywords Metabolomics Breast Cancer Liquid chromatography - mass spectrometry NMR techniques Principal component analysis Squares - discriminant analysis
CLC R737.9
Type Master's thesis
Year 2011
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The experiment: initially established based on 1HNMR technology and the UPLC-Q/TOF (LC-MS) techniques metabonomics method, and the method is applied in breast cancer patients with endogenous metabolite changes, find breast cancer diagnostic markers. On this basis, a preliminary comparison of the chemotherapy regimen AC (doxorubicin cyclophosphamide) in serum of breast cancer patients before and after intervention metabolite changes observed in vivo in patients after chemotherapy intervention endogenous metabolites in serum trends. Methods: 1. Study subjects included: Select examination results of 11 cases of normal healthy people as a control group; selected met the inclusion criteria, 16 patients with newly diagnosed breast cancer patients as the experimental group. (2) Specimen collection: 11 normal female blood collection, separation and collection of serum; experimental group, 16 cases were collected from breast cancer patients before chemotherapy and after a course of chemotherapy, blood, separating and collecting 3. Establish technology-based serum 1HNMR metabolomics analysis methods, including experimental methods and pattern recognition method of choice. Normal female and established methods and experimental groups before chemotherapy, serum metabolomics analysis to identify differences in serum metabolites; while the method is applied to breast cancer patients before and after AC program interventions serum metabolites change. Through the literature search and data queries to identify differences in serum. 4 Establish LC-MS-based techniques (UPLC-Q/TOF) technology serum metabolomics analysis platform and established method normal female and experimental groups before chemotherapy, serum metabolomics analysis to identify differences; while Methods used in AC program interventions before and after breast cancer serum metabolites change. 1HNMR technical studies and compared. 5 Data analysis: Statistical analysis through a variety of methods to analyze massive amounts of data generated, the analysis process include: normalization, revised 80% rule, data sets and data partitioning method for scaling the data set preprocessing; through the data Analysis: The use of principal component analysis (principal componentanalysis, PCA), orthogonal partial least squares discriminant analysis (OrthogonalPartial Least-Squares Discriminant Analysis, OPLS-DA) for pattern recognition analysis; according to the model of variable importance factor (VIP value), non-parametric test results and P values ??of screening potential markers. Results: 1. Establish a 1H-NMR serum metabolomics methods, healthy women and breast cancer patients before chemotherapy, chemotherapy both groups before and after intervention by metabolomics methods are well distinguished, and find the corresponding 16 breast tumor markers. Lactic acid (lactate, 1.33,4.12), valine (valine, 0.99/1.04), N-acetyl-glycoprotein (N-acetylgly-coproteins, 2.04), glutamine (glutamine, 2.41), phosphorylcholine / choline (phosphocholine / cholie, 3.22), glucose (a-glucose and B-glucose ,3.40-3 .90,4.66,5.22), tyrosine (tyrosine, 6.87,7.17), histidine (histidine), LDL / VLDL, not saturated fatty acids and other peaks increase or decrease performance. (2) establish a UPLC-Q/TOF-MS serum metabolomics research methods, found that choline and phenylalanine, isoleucine in normal subjects and patients with breast cancer before chemotherapy, there are different, in which the breast after chemotherapy cancer serum choline class has to change unit normal trend. 1HNMR technology resulting UPLC-Q/TOF markers and markers obtained substantially different, but similar results packet clustering. Conclusion: ~ 1HNMR technology and UPLC-Q/TOF technology as used metabolomics analysis tools found in markers have a certain complementarity. Through the normal subjects and patients with breast cancer before and after chemotherapy metabolomics studies, got the three metabolic profiling and found this article established NMR techniques and UPLC-Q/TOF technology-based metabonomics method was applied to breast cancer patients Serum good clustering results.

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