Proton Nuclear Magnetic Resonance Spectroscopy-based Metabonomic Research On Human Pancreatic Cancer Tissue

ObjectiveAnalysis the metabonomic differences of pancreatic cancer tissue, paracancerous tissue and normal pancreatic tissue by nuclear magnetic resonance hydrogen spectrum,analysis the possible mechanism and its clinical significance,to find potential diagnostic biomarker and therapeutic target for pancreatic cancer.It may lay the foundation for finding the potential tumor metabolic markers.Methods1、According to the following inclusive criteria about tumour group, collecting the tissue specimen of pancreatic tumors including 32 cases of pancreatic ductal adenocarcinoma tissue, 16 cases of paracancerous tissue.At the same period, collecting the tissue specimen of 27 cases of normal pancreatic tissue according to the following inclusive criteria about control group. All the tissue specimens were stored in eighty degree below zero. 2 、 The tissue was weighed,grinded,and then extracted by chloroform / methanol / water extraction, the water soluble extract became powder by 24 hours of freeze drying. 3、The powder was dissolved in the NMR detection liquid,and was detected in Varian NMR System 500 MHz NMR for acquisition of nuclear magnetic resonance spectrum. 4、Nuclear magnetic resonance spectrum was processed with the Fourier transform, phase and baseline adjustment,and improve the SNR by Top Spin software(V3.0) and calibration the spectra in bimodal 1.33 ppm of lactic acid, calibration interval is 9.5-0.5 ppm, 0.002 ppm width, integral normalized data into Excel data format, used for pattern recognition analysis.5、The data was processed by SIMCA-P+ software, and principal component analysis and partial least squares discriminant analysis and other statistical analysis were used. 6、Evaluated the stability and effectiveness of models for further OPLS-DA analysis and Pearson correlation coefficient to detect significant differences between endogenous metabolism of judgment, the tumor group and the control group.Results1、The data of pancreatic ductal adenocarcinoma tissue and normal pancreatic tissue have good clustering in PCA. Discrimination model is established. By the PLS-DA cross validation and permutation test proving, the model is correct. 2、The metabolic analysis revealed higher of 3-Hydroxybutyrate、Formate、Glutamate、Lactate、 Nicotinamide、Taurine, and lower of 3-Methylhistidine、3-Methylxanthine、4-Hydroxybenzoate、 Acetylcholine、Adenine、 Adenosine 3’,5’-diphosphate、 Alanine、AMP/ADP、 Adenosine monophosphate、Aspartate、 Adenosine Triphosphate、 Betaine、Ethanolamine、Fumarate、Glutamine、Glycine、Glycerolphosphocholine 、 Hypoxanthine 、 Inosine 、 Lysine 、 myo-Inositol 、1-Methylnicotinamide、N-Acetylaspartate、Nicotinamide adenine dinucleotide、Phenylalanine、Pyruvate、Sarcosine、Succinate、Threonine、 Tryptophan、Tyrosine、 Uracil、 Uridine、Uridine diphosphate Glucose、Xanthine in the pancreatic cancer tissue compared to that of normal pancreatic tissue. 3、The data of paracancerous tissue and cancerous tissue,paracancerous tissue and normal tissue, have no good clustering in PCA. Discrimination model is established. By the PLS-DA cross validation and permutation test proving, the model is not correct.Conclusion1、Compared with normal pancreatic tissue,there is an obvious metabolic difference in pancreatic ductal adenocarcinoma tissue.It is not only involve energe metabolism but also the metabolism of lipid, amino acids and nucleic acid.2、It may help to study the mechanism of the occurrence and development of pancreatic cancer by nuclear magnetic resonance hydrogen spectrum and PCA/PLS – DA. 3、To paracancerous tissue and cancerous tissue,paracancerous tissue and normal tissue,it is not obvious metabolic difference.