Stemness Potency and Structural Characteristics of Thyroid Cancer Cell Lines

Thyroid cancer is the most common endocrine malignancy, and its incidence is increasing rapidly worldwide [1], [2]. Papillary (PTC) (85%), follicular (FTC) (12%), and anaplastic (ATC) (<3%) thyroid carcinomas are derived from the follicular epithelium. PTC and FTC are usually treated similarly. PTC, on the other hand, has a low advancement rate, radioactive iodine ablation is effective in therapy, and it has a good prognosis [3]. ATCs are undifferentiated cancers, and they have a fatal progression due to various treatment regimens. Surgery is the first-line treatment option for patients with differentiated thyroid cancer[4].

Cancers are often caused by cancer cells that have the ability to self-renew and differentiate into multiple cell lineages. Cancer stem-like cells (CSC) or cancer initiating cells are cancer cells with stem-like characteristics. CSCs project cancer initiation, progression, and dissemination [5]. Kruppel-like factor (KLF4) and Stage-spesific embryonic antigen 1 (SSEA-1) are types of stem cell markers. KLF4 is in charge of maintaining embryonic stem cells, self-renewal, and pluripotency [6]. This protein is a zinc-finger transcription factor that belongs to the 'kruppel' family. It is essential for the control of the cell cycle, epidermal growth, and the maintenance of pluripotency [7]. Cytoplasmic KLF4 organizes cytoskeletal organization[8]. Altered KLF4 expression has been linked to a variety of malignancies, including colorectal cancer, lung cancer, gastric cancer, and urothelial cancer [9], [10], [11], [12]. However, the relevance of KLF4 expression in thyroid cancer remains unknown. SSEA-1 is another cancer stem cell marker [13], is rarely expressed in normal tissue cells. It is classified as a biomarker for thyroid CSCs and embryonic stem cells [14]. SSEA-1, which is an adhesion molecule composed of a carbonhydrate epitope with a galactose N-acetylglucoseamine fucose linkage, plays an important role in migration and adhesion of cells in the developing embryo. Besides, it is one of the pluripotent stem cell markers [13]. SSEA-1 is classified as a kind of oncofetal antigen in thyroid tissues [15], [16].

During cancer migration and metastasis, cytoskeletal alterations have been recorded. The study of cytoskeletal components and cell shape has the potential to have a substantial influence on cancer development. Filamentous actin (F-actin) is a component of the cellular cytoskeleton that is involved in cell movement and tissue remodeling [17], [18]. Actin interacts with a variety of membrane proteins, affecting cell adhesion, migration, polarity, and the sensitivity to extracellular signals[19]. An irregular actin architecture has been associated with a high metastatic potential in cancers such as colon cancer, melanoma, and fibrosarcoma. Actin architecture has been reported to indicate the status of differentiation of thyroid cancer cells [20]. Scanning electron microscopy (SEM) is an effective technique used for a long time in cell culture systems besides the light microscopy techniques to provide typically three dimensional visualization of cell surface morphology with higher resolution [21], [22], [23]. There is a limited research about Scanning Electron Microscopy-Energy Dispersive Spectroscopy (SEM-EDS) that shows the elemental composition into region of interest [24], [25], [26].

Here, we aim to evaluate stem cell markers in PTC, FTC, and ATC cell lines to normal thyroid cells with the aid of morphological features. We investigated the intensity and distribution of F-actin protein throughout the cell and, as well as the structural and elemental features of these cell lines. This study might assist in the development of therapeutic targets for cancer stem-like cells as well as the understanding of treatment resistance mechanisms.

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