ΔNP63β geninin T98G hücreleri üzerindeki fenotipik ve genotipik özelliklerinin araştırılması
Özet
P63 is described as a member of the p53 family. Although p63 resembles to p53 as structural and functional, it is different from the p53 because it has different isoforms. Although there are studies related with p63 isoforms, ΔNp63β that is an isoform of p63 are not presented in literature in detail. In this thesis the phenotypic and genetic effects of ΔNp63β on the T98G cell line was investigated. P63 is known as tumor suppressor protein. For this reason, to see the functions of ΔNp63β on continuous cell lines pRetroX-Tet-On vector system, which induced by the doxacillin, was used. First rtTA expressing vector that is part of the pRetroX-Tet-On vector system was established on the T98G cell lines with the presence of neomycine selection. Thereafter, ΔNp63β cDNA cloned Retropur Tight retroviral vector prepared and infected the T98G cell lines containing pRetroX-Tet-On vector system and continuous cell lines were obtained with the puromycin selection. Then, the functions of the ΔNp63β on the T98G cell line were investigated. Although TA p63s is described as tumor suppressor and ΔN p63 isoforms is described as oncogene in the limited number of literature, in this study, overexpression of ΔNp63β in the T98G cell lines killed cell dramatically. Interestingly, It was determined that cell death is due to the necrosis not as expected as due to apoptosis. Genetic effects of ΔNp63β was investigated basis on the expression analysis of the p53 target genes. It was found that while ΔNp63β has no effects on the apoptosis-related genes, it has changed the expression of the anti-apoptotic genes. In addition the apoptosis related genes, Interferon regulated genes which are regulated by p53 and have roles in cell defence were investigated. Results indicates that Expressions of IFN regulated genes showed similar patterns to the p53 results. Although ΔNp63β overexpression increased the some of the oncogenite releated genes, cell died dramatically. Further experiements are needed to explore the causing of cell death.