Master Degree / Yüksek Lisans Tezleri

Permanent URI for this collectionhttps://hdl.handle.net/11147/3008

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Subject: search.filters.subject.Connexin 32

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  • Master Thesis
    Time Dependent Expression and Localization of Connexin 32: Implication in Epithelial To Mesenchymal Transition of Mammary Epithelial Mcf10a and Triple Negative Breast Cancer Mda Mb 231 Cells
    (01. Izmir Institute of Technology, 2020) Ünal, Yağmur Ceren; Meşe Özçivici, Gülistan; Meşe Özçivici, Gülistan; 01. Izmir Institute of Technology; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science
    Breast cancer is the most frequent and the second leading cause of cancer-related deaths among women worldwide. Epithelial to mesenchymal transition (EMT) is critical driving force in metastasis. Connexins as a basic subunit of gap junctions indicate critical roles in regulation of EMT. In addition to Cx26 and Cx43, Cx32 is associated with breast cancer and elevated levels of Cx32 has been reported in lymph node metastasis compared to primary breast cancer while the role of Cx32 in breast cancer is still elusive. Here we aimed to shed light on the effect of Cx32 on breast cancer. Our study suggested that Cx32 acquired mesenchymal morphology and decreased proliferation in MCF10A cells but not in MDA MB 231 cells. To further elucidate whether Cx32 indicate these changes through EMT, EMT markers were examined and subsequently it was revealed that Cx32 expression was strongly correlated with increased E-cadherin and Vimentin in MCF10A cells while decreased E-cadherin and Snail in MDA MB 231 cells. Importantly majority of Cx32 did not localize to the plasma membrane and indicated dynamic changes in a day dependent manner in both MCF10A and MDA MB 231 cells. Moreover, day dependent expression and localization of Cx32 revealed strong correlation with Zeb2 expression in MCF10A cells. In conclusion, Cx32 indicated differential effects in regulation of EMT between MCF10A and MDA MB 231 cells. It was the first time that the role of Cx32 on EMT was investigated in breast cancer and differential localization of Cx32 was identified.
  • Master Thesis
    Comparison of Connnexin32 Expression and Function Between Mcf10a Normal Breast and Mda-Mb Breast Cancer Cell Lines
    (Izmir Institute of Technology, 2017) Meşe Özçivici, Gülistan; Meşe Özçivici, Gülistan; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Breast 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.