Metabolomik Ve Fluksomik Çalışmalar ile Kolon Kanserinde Kemoterapötiklere Karşı Direncin in Vitro Değerlendirilmesi

Abstract

Özen G., In Vitro Analysis of Resistance to Chemotherapeutics in Colon Cancer by Metabolomic and Fluxomic Studies, Hacettepe University Graduate of Health Sciences, Analytical Chemistry Program, Ph.D. Thesis, Ankara, 2019. Today's innovative technologies permits to perform complex analyses like omics (genomics, transcriptomics, proteomics, metabolomics and fluxomics). Detailed information obtained from omics analysis has great potential to understand the mechanism of diseases, to facilitate early diagnosis, to choose individual treatment strategies and to assess their effectiveness. Characterization of metabolic phenotype requires simultaneous and comprehensive analysis of a large number of metabolites and their turnover rates. Within the scope of thesis studies, changes in phenotype of human colon adenocarcinoma cell lines (Caco-2) which are in vitro models of colorectal cancer and human fetal colon cell lines (FHC) based on resistance devolepment to chematoratpotics were investigated by metabolomic, lipidomic and fluxomic analyzes. 18O isotope were used as a marking agent in fluxomic analyses. GC-MS and LC-qTOF-MS analytical instruments were used in the analysis of metabolomics, lipidomic and fluxomic. As a result of these analyzes, the more than 300 metabolites / lipids and 18O labeling percentages of 38 metabolites were calculated. In thesis studies, incorporation 18O in to Krebs cycle’s metabolites, amino acids, fatty acids and oligophosphonucleotides were determined fort he first time. The pathways analyses based on the significantly altered metabolits (p<0.05) showed that alteration in the malate-aspartate shuttle, amino acid, ammonia metabolism, glucose-alanine and urea cycles pathways changed depending on the resistance. The data obtained from comprehensive omics (metabolomics, lipidomics and fluxomics) studies of these cell lines showed differences in cell dynamics of resistance to chemotherapeutics.

Keywords: Fluxomics, Metabolomics, Lipidomics, ,LC-qTOF-MS, GC-MS, Caco-2, colon adenocarcinoma Supported by: TUBİTAK 1001 Support Project (116Z292)