Master Degree / Yüksek Lisans Tezleri

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

Browse

Filters

2014 - 20208

Settings

Department: search.filters.department.Thesis (Master)--İzmir Institute of Technology, Molecular Biology and GeneticsSubject: search.filters.subject.Cancer cells

Search Results

Now showing 1 - 8 of 8
  • 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
    Investigating Oncogenic Role of Sema6d in Breast Cancer Cells
    (Izmir Institute of Technology, 2019) Günyüz, Zehra Elif; Yalçın Özuysal, Özden; Yalçın Özuysal, Özden; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Breast cancer, the most commonly diagnosed cancer type and the leading cause of cancer-associated deaths, is the major health issue among women worldwide. In many cancer types, the expression of the semaphorins and their receptors such as plexins and neuropilins are dysregulated. SEMA6D is a member of class-6 family transmembrane semaphorin proteins and acts through Plexin-A1 receptor. It was previously shown that overexpression of SEMA6D in breast cancer cell line MCF-7 leads to a reduction in proliferation and an increase in migration. On the other hand, in the MDA-MB-231 breast cancer cell line, overexpression of SEMA6D had no significant effect on proliferation but enhanced migration. In this study, we aimed to analyze the effects of SEMA6D overexpression in normal breast cell line MCF10A and investigate the invasive behavior and transformation capacity of SEMA6D overexpressing breast cancer cell lines. We demonstrated that overexpression of SEMA6D leads to elevated proliferation, viability and migration in MCF10A cells, whereas it did not trigger their anchorage-independent growth. On the other hand, MDA-MB-231 and MCF7 cells stably expressing SEMA6D showed reduced colony formation in the soft-agar assay. Furthermore, the invasiveness of MDA-MB-231 cells was elevated with SEMA6D overexpression, whereas SEMA6D overexpression did not stimulate the invasiveness of MCF-7 cells through matrigel microenvironment, whereas slightly trigger invasion through bone microenvironment. In conclusion, SEMA6D overexpression has cell-specific effects on breast cancer. The exact role of SEMA6D in breast cancer development remains undefined and must be further investigated.
  • Master Thesis
    Determination of Therapeutic Potential of Luteolin for Acute Lymphoblastic Leukemia Cells
    (Izmir Institute of Technology, 2019) Gürler, Sevim Beyza; Baran, Yusuf; Baran, Yusuf; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Acute lymphoblastic leukemia (ALL) is a hematologic malignancy characterized by increased level of immature lymphoblasts in bone marrow and peripheral blood. The developments of lymphoblasts are genetically/epigenetically inhibited. One of the most common genetic abnormalities in ALL is BCR/ABL translocation which regulates the several pathways related to proliferation, anti-apoptotic and drug resistance through its aberrant tyrosine kinase activity. Although the current treatment strategies include targeting BCR/ABL via tyrosine kinase inhibitors; complete remission, overall survival and mortality of Ph+ ALL patients are still worse as compared to Ph- ALL patients. Therefore, new strategies combined with current treatments are needed for Ph+ ALL patients who are qualified as high risk group of ALL. Different studies showed thatluteolin has anti-cancer and anti-tumor effects on wide range cancer types including breast, colon, lung cancer except ALL in both in vitro and in vivo. In this study, the dose and time dependent cytotoxic, apoptotic and cytostatic effects of luteolin on Philadelphia chromosome +ALL cells were determined for the first time. Besides, the effect of luteolin on cell growth and proliferation of two different healthy cell lines was shown. Moreover, the effect of luteolin on bioactive sphingolipids genes which regulate the several pathways including cell proliferation, apoptosis, drug resistance and senescence in cell was determined in Ph positive ALL cells for the first time. As a consequence, luteolin has cytotoxic, apoptotic and cytostatic effects on Ph positive ALL cells and bioactive sphingolipids genes are regulated in this therapeutic potential by luteolin.
  • Master Thesis
    Analysis of Tnfrsf10b-As Long-Noncoding Rna's Effects on Various Cancer Cell Properties
    (Izmir Institute of Technology, 2019) Alkan, Ayşe Hale; Akgül, Bünyamin; Akgül, Bünyamin; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Long noncoding RNAs (lncRNAs) being longer than 200 nucleotides constitute a different class of RNA molecules. Several studies indicated that they have regulatory role in cellular processes including cancer development. Some of them have exclusively high expression in particular cancer types and regulate certain cancer cell properties. This renders them potential biomarker or therapeutic target in cancer. In this study, effects of a candidate lncRNA TNFRSF10B-AS and lncCAMTA1 on cancer cell properties were investigated. Candidate lncRNAs from Doxorubicin, Fas mAB, TNF-alpha and Cisplatin treated HeLa cell line were chosen and their expression level was measured in different cell lines including healthy (BEAS2B and MCF10A), metastatic (H1299 and MDA-MB- 231) and non-metastatic cell lines (A549 and MCF-7) by qPCR. From a few candidates lncCAMTA1 and TNFRSF10B-AS were selected for further analysis. qPCR results obtained from comparison of different cancer cell lines showed that their expression differs at least in one comparison of cell lines. TNFRSF10B-AS silencing decreased proliferation of HeLa cells. lncCAMTA1 was silenced or overexpressed in HeLa cells but phenotypic effect couldn’t be detected by apoptosis and cell proliferation assay. Additionally, phenotypic effect also couldn’t be observed in other cell lines when TNFRSF10B-AS was silenced.
  • Master Thesis
    Determination of Therapeutic Effects of Multifunctional Micelle-Based Nanocarriers on Breast Cancer Cells
    (Izmir Institute of Technology, 2019) Ulu, Gizem Tuğçe; Baran, Yusuf; Ulu, Gizem Tuğçe; Baran, Yusuf; 01.01. Units Affiliated to the Rectorate; 04.03. Department of Molecular Biology and Genetics; 01. Izmir Institute of Technology; 04. Faculty of Science
    Breast cancer is the most common and frequent cause of death among women composed to all types of cancer. Current treatment protocols do not provide complete cure or selective drug delivery while targeted therapy can provide an important avenue for successful treatment of breast cancer. In this study, therapeutic effects of drug-conjugated nanocarrier system with enhanced stability and double moiety pH-sensitivity on breast cancer (SKBR-3- HER-2- positive), normal breast epithelial (MCF-10A, HER-2-negative) and chronic myeloid leukemia (K562, HER-2-negative) cells were determined. With this approach, SKBR-3 cells were targeted by single nanocarriers having selectivity with unused peptide ligand (HER-2), stability with cross-linking of core moiety, and cleavage by two sites of pHeffect and drug release properties. After physicochemical characterization of micellebased nanocarriers, cytotoxic, apoptotic and cytostatic effects of doxorubicin conjugated micelles were determined. Doxorubicin conjugated micelles with HER-2 peptide (DOX-HER-2-NCs) had more cytotoxic effects on HER-2 positive cells. Additionally, intracellular amounts of doxorubicin is higher in SKBR-3 cells with applied DOX-HER-2-NCs as determined by fluorescence imaging. The apoptosis rate was increased on SKBR-3 at 50% cell growth inhibition (IC50) as determined by Annexin-V/Propidium iodide double staining. However, there was not any significant change in loss of mitochondrial membrane potential. Additionally, DOX-HER-2-NCs resulted in cell cycle arrest at G2/M-phase in response to IC50 value. Besides, protein level of Bcl-2 did not change while protein level of Bax and Caspase-3 were increased as determined by Western Blotting. This project provides novel and more effective treatment of breast cancer by using multifunctional properties of nanocarriers.
  • Master Thesis
    Cyr61-Notch Interaction During Epithelial-To Transition, Migration and Invasion in Breast Cancer Cells
    (Izmir Institute of Technology, 2017) İlhan, Mustafa; Yalçın Özuysal, Özden; Yalçın Özuysal, Özden; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Notch signaling is one of the major pathways involved in development and tumorigenesis. Activated Notch is correlated with increased migration, invasion and epithelial-to-mesenchymal-transition (EMT) in breast cancer. However, mechanism of Notch-mediated cancer progression is poorly understood. CYR61 is a secreted protein and its upregulation is also related to increased breast tumorigenesis and EMT. Microarray analyses revealed that CYR61 was differentially expressed in response to Notch activation in breast epithelial cells. We hypothesized that CYR61 is a downstream mediator of Notch during EMT, migration and invasion. To test whether Notch requires CYR61 during EMT, migration and invasion, two complementary approaches were followed: (i) CYR61 expression was silenced by using shRNA in MCF10A epithelial breast cell line in the presence of Notch activation, (ii) CYR61 was over-expressed in MDA-MB-231 cancer breast cell line in the absence of Notch activity. Then, expression of EMT markers was analyzed in mRNA and protein levels via RT-qPCR and immuno-blotting, respectively. Migration and invasion ability of cells were investigated by wound healing assay and a lab-on-a-chip-system, respectively. Here, it was shown that CYR61 inhibition decreased Notch-induced EMT, migration and invasion of MCF10A and CYR61 overexpression in the absence of Notch activity partially rescued Notch-mediated invasion in MDA-MB-231 cell lines. Our findings suggest that CYR61 may act in downstream of Notch and is regulated by Notch. When we consider importance of CYR61 in Notch-induced EMT and cancer progression, targeting CYR61 may hold promise to develop novel strategies for treatment of breast cancer in early stages.
  • 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.
  • Master Thesis
    Cell Adhesion on Nanomater Scale Fibronectin Patterns: a Comparision of Breast Cancer Cells and Normal Breast Epithelial Cells
    (Izmir Institute of Technology, 2014) Horzum, Utku; Pesen Okvur, Devrim; Pesen Okvur, Devrim; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    Cell adhesion to extracellular matrix is an important process for both health and disease states. Surface protein patterns are topographically flat, and do not introduce other chemical, topographical or rigidity related functionality and, more importantly, that mimic the organization of the in vivo extracellular matrix are desirable. Previous work showed that vinculin and cytoskeletal organization are modulated by the size and shape of surface nanopatterns. However, a comparative and quantitative analysis on normal and cancerous cell morphology and focal adhesions as a function of micrometer scale spacings of protein nanopatterns was absent. Here, electron beam lithography was used to pattern fibronectin (FN) nanodots with micrometer scale spacings on a K-casein background (single active) on indium tin oxide (ITO) coated glass which, unlike silicon, is transparent and thus suitable for many light microscopy techniques. Exposure times were significantly reduced using the line exposure mode with micrometer scale step sizes. Micrometer scale spacings of 2, 4, 8 microns and gradients between FN nanodots modulated cell adhesion for both breast cancer and normal mammary epithelial cells, through modification of cell area, cell symmetry, actin organization, focal adhesion number, size and circularity under both static and flow conditions. Overall, cell behavior was shown to shift at the apparent threshold of 4 μm spacing. Results showed that there were significant differences in terms of cell adhesion between breast cancer and normal mammary epithelial cells: Breast cancer cells exhibited a more dynamic and flexible adhesion profile than normal mammary epithelial cells.