Browsing by Author "Ozdil, Berrin"

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Autophagy and Mtor Pathways in Mouse Embryonic Stem Cell, Lung Cancer and Somatic Fibroblast Cell Lines
(Wiley, 2019) Oltulu, Fatih; Kocaturk, Duygu C.; Adali, Yasemin; Ozdil, Berrin; Acikgoz, Eda; Gurel, Cevik; Aktug, Huseyin
Embryonic developmental stages and regulations have always been one of the most intriguing aspects of science. Since the cancer stem cell discovery, striking for cancer development and recurrence, embryonic stem cells and control mechanisms, as well as cancer cells and cancer stem cell control mechanisms become important research materials. It is necessary to reveal the similarities and differences between somatic and cancer cells which are formed of embryonic stem cells divisions and determinations. For this purpose, mouse embryonic stem cells (mESCs), mouse skin fibroblast cells (MSFs) and mouse lung squamous cancer cells (SqLCCs) were grown in vitro and the differences between these three cell lines signalling regulations of mechanistic target of rapamycin (mTOR) and autophagic pathways were demonstrated by immunofluorescence and real-time polymerase chain reaction. Expressional differences were clearly shown between embryonic, cancer and somatic cells that mESCs displayed higher expressional level of Atg10, Hdac1 and Cln3 which are related with autophagic regulation and Hsp4, Prkca, Rhoa and ribosomal S6 genes related with mTOR activity. LC3 and mTOR protein levels were lower in mESCs than MSFs. Thus, the mechanisms of embryonic stem cell regulation results in the formation of somatic tissues whereas that these cells may be the causative agents of cancer in any deterioration.
Cd133+/Cd44+prostate Cancer Stem Cells Exhibit Embryo-Like Behavior Patterns
(Elsevier Gmbh, 2021) Acikgoz, Eda; Soner, Burak Cem; Ozdil, Berrin; Guven, Mustafa
Cancer stem cells (CSCs), which act as an important bridge between cancer formation and embryonic development, represent a small population associated with tumor initiation, drug resistance, metastasis and recurrence. CSCs have the ability to form spheroids in three-dimensional culture systems. Tumor spheroids derived from CSCs with symmetric and asymmetric division patterns were found to contain highly heterogeneous cell groups. The biological behavior patterns which some CSCs display serve as an important bridge between cancer formation and embryonic development. The cell population in the DU-145 prostate cancer cell line with surface markers CD133+/CD44+ was isolated by FACS. Prostate spheroids were formed by using agarose-coated plates. The morphological characteristics of the cell population within spheroid structure and the expression of Ki-67 and Caspase-3 were investigated by histochemical methods. In this study, we observed that CD133+/CD44+ prostate CSCs form different spheroid structures as well as normal spheroid structures: i) some spheroid structures formed with a highly transparent zone on the outer part of the spheroid, in addition to the normal spheroidal zones and ii) spheroidal structures obtained from prostate CD1334+/CD44+ CSCs that share the same microenvironment are hollow spheres similar to the blastula-like structure in the embryo. These spheroidal structures exhibiting embryo-like properties indicate that the expression of embryonic factors might be reiterated in CSCs. Further investigation of the formation mechanism of the transparent zone and the hollow sphere will shed light on the embryonic origin of prostate cancer and the design of new therapeutic strategies.
Differences and Similarities Between Colorectal Cancer Cells and Colorectal Cancer Stem Cells: Molecular Insights and Implications
(Amer Chemical Soc, 2023) Erisik, Derya; Ozdil, Berrin; Acikgoz, Eda; Asker Abdikan, Cemile Sinem; Yesin, Taha Kadir; Aktug, Huseyin
Malignant tumors are formed by diverse groups of cancercells.Cancer stem cells (CSCs) are a subpopulation of heterogeneous cellsidentified in tumors that have the ability to self-renew and differentiate.Colorectal cancer (CRC), the third most frequent malignant tumor,is progressively being supported by evidence suggesting that CSCsare crucial in cancer development. We aim to identify molecular differencesbetween CRC cells and CRC CSCs, as well as the effects of those differenceson cell behavior in terms of migration, EMT, pluripotency, morphology,cell cycle/control, and epigenetic characteristics. The HT-29 cellline (human colorectal adenocarcinoma) and HT-29 CSCs (HT-29 CD133(+)/CD44(+) cells) were cultured for 72 h. The levelsof E-cadherin, KLF4, p53, p21, p16, cyclin D2, HDAC9, and P300 proteinexpression were determined using immunohistochemistry staining. Themigration of cells was assessed by employing the scratch assay technique.Additionally, the scanning electron microscopy method was used toexamine the morphological features of the cells, and their peripheral/centralelemental ratios were compared with the help of EDS. Furthermore,a Muse cell cycle kit was utilized to determine the cell cycle analysis.The HT-29 CSC group exhibited high levels of expression for E-cadherin,p53, p21, p16, cyclin D2, HDAC9, and P300, whereas KLF4 was foundto be high in the HT-29. The two groups did not exhibit any statisticallysignificant differences in the percentages of cell cycle phases. Theidentification of specific CSC characteristics will allow for earliercancer detection and the development of more effective precision oncologyoptions.
Differences and Similarities in Biophysical and Biological Characteristics Between U87 Mg Glioblastoma and Astrocyte Cells
(Springer, 2024) Ozdil, Berrin; Calik-Kocaturk, Duygu; Altunayar-Unsalan, Cisem; Acikgoz, Eda; Oltulu, Fatih; Gorgulu, Volkan; Aktug, Huseyin
Current cancer studies focus on molecular-targeting diagnostics and interactions with surroundings; however, there are still gaps in characterization based on topological differences and elemental composition. Glioblastoma (GBM cells; GBMCs) is an astrocytic aggressive brain tumor. At the molecular level, GBMCs and astrocytes may differ, and cell elemental/topological analysis is critical for identifying potential new cancer targets. Here, we used U87 MG cells for GBMCS. U87 MG cell lines, which are frequently used in glioblastoma research, are an important tool for studying the various features and underlying mechanisms of this aggressive brain tumor. For the first time, atomic force microscopy (AFM), scanning electron microscopy (SEM) accompanied by energy-dispersive X-ray spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS) are used to report the topology and chemistry of cancer (U87 MG) and healthy (SVG p12) cells. In addition, F-actin staining and cytoskeleton-based gene expression analyses were performed. The degree of gene expression for genes related to the cytoskeleton was similar; however, the intensity of F-actin, anisotropy values, and invasion-related genes were different. Morphologically, GBMCs were longer and narrower while astrocytes were shorter and more disseminated based on AFM. Furthermore, the roughness values of these cells differed slightly between the two call types. In contrast to the rougher astrocyte surfaces in the lamellipodial area, SEM-EDS analysis showed that elongated GBMCs displayed filopodial protrusions. Our investigation provides considerable further insight into rapid cancer cell characterization in terms of a combinatorial spectroscopic and microscopic approach.
The Effect of Extracellular Matrix on the Differentiation of Mouse Embryonic Stem Cells
(Wiley, 2020) Ozdil, Berrin; Guler, Gunnur; Acikgoz, Eda; Kocaturk, Duygu Calik; Aktug, Huseyin
Embryonic stem cells (ESCs) are promising research materials to investigate cell fate determination since they have the capability to differentiate. Stem cell differentiation has been extensively studied with various microenvironment mimicking structures to modify cellular dynamics associated with the cell-extracellular matrix (ECM) interactions and cell-cell communications. In the current study, our aim was to determine the effect of microenvironmental proteins with different concentrations on the capacity and differentiation capability of mouse ESCs (mESCs), combining the biochemical assays, imaging techniques, Fourier transform infrared (FTIR) spectroscopy, and unsupervised multivariate analysis. Based on our data, coating the surface of mESCs with Matrigel, used as an acellular matrix substrate, resulted in morphological and biochemical changes. mESCs exhibited alterations in their phenotype after growing on the Matrigel-coated surfaces, including their differentiation capacity, cell cycle phase pattern, membrane fluidity, and metabolic activities. In conclusion, mESCs can be stimulated physiologically, chemically, or mechanically to convert them a new phenotype. Thus, identification of ESCs' behavior in the acellular microenvironment could be vital to elucidate the mechanism of diseases. It might also be promising to control the cell fate in the field of tissue engineering.
Spectroscopic and Microscopic Comparisons of Cell Topology and Chemistry Analysis of Mouse Embryonic Stem Cell, Somatic Cell and Cancer Cell
(Elsevier Gmbh, 2021) Ozdil, Berrin; Calik-Kocaturk, Duygu; Altunayar-Unsalan, Cisem; Acikgoz, Eda; Gorgulu, Volkan; Uysal, Aysegul; Aktug, Huseyin
While embryonic stem cells and cancer cells are known to have many similarities in signalling pathways, healthy somatic cells are known to be different in many ways. Characterization of embryonic stem cell is crucial for cancer development and cancer recurrence due to the shared signalling pathways and life course with cancer initiator and cancer stem cells. Since embryonic stem cells are the sources of the somatic and cancer cells, it is necessary to reveal the relevance between them. The past decade has seen the importance of interdisciplinary studies and it is obvious that the reflection of the physical/chemical phenomena occurring on the cell biology has attracted much more attention. For this reason, the aim of this study is to elementally and topologically characterize the mouse embryonic stem cells, mouse lung squamous cancer cells, and mouse skin fibroblast cells by using Atomic Force Microscopy (AFM), X-ray Photoelectron Spectroscopy (XPS) and Scanning Electron Microscopy (SEM) supported with Electron Dispersive Spectroscopy (EDS) techniques in a complementary way. Our AFM findings revealed that roughness data of the mouse embryonic stem cells and cancer cells were similar and somatic cells were found to be statistically different from these two cell types. However, based on both XPS and SEM-EDS results, surface elemental ratios vary in mouse embryonic stem cells, cancer cells and somatic cells. Our results showed that these complementary spectroscopic and microscopic techniques used in this work are very effective in cancer and stem cell characterization and have the potential to gather more detailed information on relevant biological samples.