Forming of induced cancer stem-like cells as a model of cancer cell


Thesis Type: Postgraduate

Institution Of The Thesis: Bursa Uludağ University, Fen Bilimleri Enstitüsü, Moleküler Biyoloji ve Genetik, Turkey

Approval Date: 2017

Thesis Language: Turkish

Student: SEVİL GONCA

Supervisor: Sezai Türkel

Abstract:

Yamanaka et al, achieved to induce pluripotency features in somatic cells by reprogramming in 2006. Four stemness factors (Oct3/4, SOX2, KLF4 c-MYC) were determined for reprogramming. In this study, we aimed to form induced cancer stem like cells as a cancer cell model by the transfection of the plasmid which contains four defined stemness factors by Yamanaka et al into chronic myeloid leukemia cell lines K562 and KU812. After the transfection, gen expressions of stemmnes factors SOX2 and OCT4 were shown by RT-qPCR. On the tenth day of plazmid transfection, in the KU812 cell line were observed an increase of 7230 fold in SOX2 gene expression and 1320 fold in OCT4 gene expression. In K562 cel line,Sox2 gene expression increased 92,4-fold and OCT4 gene expression increased 1,1 fold. The protein levels of the spesific markers were tested flow cytometry and immunofluorescent staining. The results showed that, the CD31, CD44 and CD105 protein expression levels increased highly significant in both transfected K562 and KU812 cell lines. Neuronal differantiation of K562 cell line were displayed using neurofilament antibody by immunoflourescent staining. After transfection of four defined factors, KU812 cells showed high telomerase activity around 4-fold compared to non-plasmids. After neuronal differentiation of K562 cells, telomerase activity was detected about 15-fold lower than that of the non-plasmids.