Ht29 Ve K562 Kanser Hücrelerinde Protein ve Metabolitlerin Analizi Için Çeşitli Analitik Yöntemlerin Geliştirilmesi

Abstract

Cancer is one of the most dangerous fatal diseases threating the human being both economically and socially. Therefore, the studies are performed on molecular level to understand cancer on molecular level and prevent the people from cancer. There are mainly two different objectives in this thesis. The first objective is to investigate the molecular mechanism of the anticancer activity of Rosmarinus officinalis L. on HT29 column cancer cells and K562/R and K562/wt leukemia cells. For this purpose, the enrichment extracts of Rosmarinus officinalis including polyphenols were appiled on in vitro cultures. The proteins were separated through two dimensional gel electrophoresis and the over- or under-expressed proteins through extracts treatment on cell lines were compared. The proteins found to be changed quantitatively on treated cells and control group were analyzed by matrixassisted laser desorption/ionization - time of flight - mass spectrometry and identified through using protein databases. The identified proteins seem to be associated with the anticancer activity according to the literature. The second objective is to adapt the XCMS, which is a metabolite profiling software working under ¨R¨ language for liquid chromatography - mass spectrometry studies, on capillary electrophoresis - mass spectrometry experiments. For this purpose, the metabolite of K562/R cell line were analyzed by using capillary electrophoresis - mass spectrometry and the electropherograms were used on metabolite profiling through the evalution of XCMS parameters for capillary electrophoresis. Some of the identified metabolites were tried to be associated with the cancer.