Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
Browse
2 results
Search Results
Master Thesis Investigating the Function of Cx26-I30n and D50y Mutations in Squamous Cell Carcinoma Cell Line Scc-25(01. Izmir Institute of Technology, 2021) Yavuz, Büşra; Meşe Özçivici, GülistanGap junctions are responsible for cell homeostasis and provide cell-cell and cell-ECM communications in multicellular organisms. Gap junction hemichannels consist of connexin (Cx) proteins which functions are crucial for human physiology. Moreover, Cx mutations are associated with various genetic diseases. For example, more than 10 missense mutations in the Cx26 gene cause keratitis-ichthyosis-deafness (KID) syndrome. Among these, Cx26-I30N and Cx26-D50Y cause to form abnormal hemichannels and allow excessive calcium influx into the cell. Furthermore, KID patients have higher squamous cell carcinoma (SCC) incidence and SCC is seen at an earlier age in these patients. Immunohistochemistry stainings have demonstrated aberrant E-cadherin and B-catenin spread in KID patients' samples. In this study, we aimed to effects of Cx26-I30N and Cx26-D50Y mutations associated with KID syndrome in human epithelial SCC cell line SCC-25. For this purpose, stable MSCV, Cx26-WT, Cx26-I30N and Cx26-D50Y cell lines were generated and RTqPCR, Western blotting, immunostaining and MTT assays were done for expression level, localization and viability analysis. For these 4 different cell conditions, a significant increase was detected at Cx26 mRNA levels, but not at protein levels. No difference was found in Cx43 level, which is abundant in epithelial tissue, and was localized in the perinuclear area like Cx26. Additionally, E-cadherin protein levels increased and their localization changed in parallel with the mutations from the perinuclear area to the plasma membrane. Moreover, significant decreases in viability were observed in cells grown in high extracellular Ca2+ medium compared to control in contrast to cells grown in Ca2+-free medium.Master Thesis Comparison of Connnexin32 Expression and Function Between Mcf10a Normal Breast and Mda-Mb Breast Cancer Cell Lines(Izmir Institute of Technology, 2017) Adak, Aslı; Meşe Özçivici, GülistanBreast cancer is one of the most prominent cancer-related deaths among females. Among many molecules, connexins have role in breast cancer. Gap junctions, formed from connexins (Cx), facilitate intercellular communication between adjacent cells. Different connexins were expressed during different stages of breast cancer. Cx32 was found both in normal pre-menopausal and tumor breast tissue samples. In lymph node metastases, elevated Cx32 level was observed compared to primary cancer. However, the role of Cx32 in breast cancer is not known but its elevation in lymph node metastasis may indicate its diverse functions in breast cancer. To verify this, in MCF10A and MDA-MB-231 cell lines, Cx32 was overexpressed. The protein localization was compared with immunostaining. In MDAMB- 231 cells, Cx32 localized in nucleus and cytoplasm, although in MDA-MB-231 Cx32-EGFP cells, Cx32 localized mostly in the cytoplasm. In MCF10A cells, Cx32 localized in nucleus, whereas Cx32 formed gap junctional plaques between MCF10A Cx32-EGFP cells. By Cx32 overexpression, gap junction coupling increased in MCF10A cells significantly, although it did not change in MDA-MB-231 cells. In both cells, hemichannel activity was not altered with Cx32 overexpression. The effects of Cx32 overexpression on cell viability demonstrated a significant decrease in MCF10A cells and an increasing trend in MDA-MB-231 cells. Furthermore, the percentage of G1 phase decreased, G2 and S phases increased in MDA-MB-231. However, Cx32 overexpression did not alter cell cycle profile of MCF10A significantly. Determination of the differential role of Cx32 in different stages of breast cancer may help to understand its diagnostic and/or therapeutic potential.