| This thesis describes application of proteomics in ovarian endometrioid ovarian carcinomas.;Chapter 2 discusses the utilization of 2-D liquid-based separation/mass mapping techniques to facilitate the analysis of protein expression between both the low- and high-stage tumors. Highly expressed proteins distinguishing low- versus high-stage OEAs were identified and validated by mass spectrometry and immunohistochemical analysis. Further, these protein profiles clearly distinguish OEAs between stages as well as from other types of ovarian cancer at the protein level.;Chapter 3 presents development of a micro separation technique, micro-CF, for analyzing two ovarian cancer cell lines MDAH-2774, and TOV-112D. Integrated microCF-nanoLC-MS/MS methods detected 700-800 proteins from ovarian cancer cell line lysates using 10[1g of samples and compared their expression between two samples using label free semi-quantitation method.;Chapter 4 describes the study of specific autoantibodies driven from pancreatic ductal adenocarcinoma using a Panc-1 cell line. Fractionated protein solutions were spotted on nitrocellulose slides and probed with total 86 sera. The data were analyzed by 4 different statistical analyses (COPA, OS, Wilcoxon, and Pam). Each statistics generated the list of differential response proteins in comparison between cancer vs normal, cancer vs pancreatitis, pancreatitis vs normal. Among identified proteins using LC-MS/MS, annxein 2, and cytoplasmic, malate-dehydrogenase, showed higher response in cancer sera against normal.;Chapter 5 presents the study on mouse model of human ovarian endometrioid adenocarcinomas using cIEF followed by tandem mass spectrometry. Mouse tumors with the p53 mutation, in addition to defects on Wnt/beta-catenin and PI3K/Pten signaling pathways, has revealed more proteins involved in signaling, and up-regulated expression of proteins in many signaling and metabolic pathways. p53 appears to play an important role in tumor progression. Also, a number of proteins detected in tumors without mutant p53 showed increased expression levels, suggesting the possibility of early detection of OEAs.;Chapter 6 describes testing a newly developed plate for MALDI-QIT-TOF-MS/MS experiments. The new plate shows increased signal compared to a stainless steel plate, higher tolerance for solvents and usability of alpha-CHCA for MS/MS. On-plate digestion method where intact proteins were digested on MALDI plate was tested and applied with real tumor samples. |
