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    Apoptotic effect of thymoquinone on OVCAR3 cells via the P53 and CASP3 activation
    (Acta Cirurgica Brasileira, 2024) Karaosmanoglu, Ozge; Kamalak, Zeynep; Ozdemir, Ilhan; Tuncer, Mehmet Cudi; Ozturk, Tamil
    Purpose: The limitations in cancer treatment and the inadequacy of classical methods have made it necessary to discover therapeutics in cancer treatment. The cytotoxicity of thymoquinone, which has quite different properties in terms of biological activities, in ovarian cancer cells, and the changes in the expression levels of apoptotic genes (p53/caspase-3 (casp-3)) were investigated. Methods: In the study, thymoquinone (5, 50, 100, 250 and 500 mu M and 24, 48, 72 hours) were applied to ovarian adenocarcinoma cancer cell line (OVCAR3), at different concentrations. Cytotoxic effect of thymoquinone on OVCAR-3 cells were analyzed by MTT method, and apoptotic and pro-apoptotic gene expression levels (p53, Casp-3) of thymoquinone in cancer cells were analyzed by quantitative real-time polymerase chain reaction. Results: Thymoquinone, whose effect has been revealed in many types of cancer, was shown to significantly reduce the viability of OVCAR3 cancer cells depending on the dose and time (p < 0.05). It was also determined that Casp-3 and p53 gene expressions increased in OVCAR3 cells. Conclusion: Considering the in-vitro cytotoxic activity and apoptotic gene expressions of thymoquinone, an important treatment agent, since it is a promising agent for the future of cancer treatment, more comprehensive studies may pave the way for its clinical use.
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    Effects of Curcumin and Doxorubicin on the Viability of Neuroblastoma Cancer Cell Line
    (2024) Ünlü, İlker; Özdemir, İlhan; Öztürk, Şamil; Tuncer, Mehmet Cudi
    Aim: Neuroblastoma has a very important place among childhood diseases, and despite all the methods used in treatment, it is very difficult to prevent neuroblastoma invasion. The number of studies showing that curcumin, the most active component of turmeric, is not toxic, is increasing day by day. In this study, the anticancer activities of curcumin (Cur), one of the important active compounds, were demonstrated in the Neuroblastoma cancer cell line (Na2B). Method: Neuroblastoma cell line was used in the study. To determine the IC50 doses of Dox and Cur, Na2B cells line were cultivated with an automatic pipette. MTT analysis was performed to analyze cell survival (viability). Inhibition levels in the cells were determined at 24 and 48 hours. Results: While the IC50 of Na2B cell proliferation was approximately 124.5 uM at the 48th hour in Dox-treated cells, the IC50 value of Cur at the 48th hour was found to be 224.6 uM. Conclusion: These results showed that Cur could be an alternative agent in the treatment of neuroblastoma, and its fewer side effects compared to other chemotherapeutic agents such as Dox would increase its preferability.
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    Gallic Acid Induces HeLa Cell Lines Apoptosis via the P53/Bax Signaling Pathway
    (Mdpi, 2024) Sari, Umut; Zaman, Fuat; Ozdemir, Ilhan; Ozturk, Samil; Tuncer, Mehmet Cudi
    Background: Cervical cancer is a type of cancer that originates from the endometrium and is more common in developed countries and its incidence is increasing day by day in developing countries. The most commonly prescribed chemotherapeutic drugs limit their use due to serious side effects and the development of drug resistance. For this reason, interest in new active ingredients obtained from natural products is increasing. This study aimed to reveal the apoptotic and antiproliferative effects of gallic acid and doxorubicin combination therapy against the HeLa cell line. Methods: We investigated the anti-cancer effects of doxorubicin and gallic acid in the human HeLa cervical cell line by using the MTT test, Nucblue staining for the identification of apoptotic cells due to nuclear condensation using fluorescent substance, and apoptotic markers P53 and Bax for the RT-PCR test. Results: The highest cytotoxic effect obtained in the study, the highest increase in apoptotic induction, and a significant difference in P53/Bax levels were seen in the gallic acid/doxorubicin combination. Additionally, it was determined that gallic acid exhibited an effective cytotoxic effect on HeLa and HaCat cells within 48 and 72 h of application. Conclusions: The obtained findings show that the gallic acid/doxorubicin combination applied to HeLa cells may be an alternative treatment against both the cytotoxic effect size and the side effects of the chemotherapy agent.
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    Histopathological Changes in The Spinal Cord Tissue of Rats Administered an Experimental Mussel Diet
    (2024) Ünlü, İlker; Özdemir, İlhan; Tuncer, Mehmet Cudi; Öztürk, Şamil
    Aim: Regional eating habits show that it causes neurodegenerative problems due to heavy metals that can accumulate in consumed foods and affect tissues such as the nervous system. Since crustaceans such as mussels feed by filtering the water, they are exposed to toxic plankton and various chemicals, especially heavy metals. Due to the limitations of experimental studies on this subject, the effects of mussel consumption on the spinal cord were investigated. Methods: In this study, histopathological changes in the spinal cord tissue of rats fed with shellfish collected from the Dardanelles were determined. The subjects were divided into two groups, and the first group was fed standard rat food for 4 weeks, and the second group was fed a mussel diet. At the end of the study, spinal cord tissue samples taken from rats were subjected to routine histopathological procedures and evaluated under a light microscope. Results: In the experimental group, a decrease in the number of neurons in the medulla spinalis and an increase in the number of astrocytes were noted. TUNEL staining showed that apoptosis occurred intensively in glial cells, but did not occur in anterior and posterior horn motor neurons. Conclusion: The findings showed that long-term mussel consumption can cause axonal damage in motor and sensory neurons and degeneration in glial cells. For this reason, it is important for health that marine diets in coastal areas are made with healthy and hygienic products.
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    Inhibitory effect of Curcumin on a cervical cancer cell line via the RAS/RAF signaling pathway
    (F Hernandez, 2025) Ozdemir, Ilhan; Zaman, Fuat; Bas, Dilek Dogan; Sari, Umut; Ozturk, Samil; Tuncer, Mehmet Cudi
    Objective. Cervical cancer has a very important place in female infertility and ranks fourth among cancers affecting women. Curcumin (CUR) is closely associated with the expression and activity of various regulatory proteins. It is also known that curcumin has preventive and therapeutic effects on various types of cancer. In this study, the anticancer activities of curcumin were demonstrated in the human cervical cancer cell line (HeLa). Methods. qRT-PCR and western blot analyses were used to evaluate mRNA and protein expression of curcumin in HeLa and immortalized human skin keratinocyte cell lines (HaCaT) (proliferation and apoptosis regulatory markers of the RAS/RAF signaling pathway). MTT analysis was performed, showing HeLa and HaCaT cell proliferation depending on the dose and duration of curcumin and doxorubicin. A wound scratch healing assay was applied to examine cell migration and invasion of HeLa after curcumin application. To determine the role of curcumin and doxorubicin in the apoptosis of HeLa cells, the mRNA levels of caspase-3 were examined by qRT-PCR. The results were analyzed with a one-way ANOVA SPSS 20.0 program. Results. CUR (IC50: 242.8 mu M) and DOX (IC50: 92.1 mu M) were determined to have the ability to inhibit the proliferation of HeLa cells and induce apoptosis over a 72-hour period and dose- dependently. Moreover, the results revealed that the mRNA and protein expression levels of RAF and RAS in HeLa cells were downregulated by CUR and DOX. Conclusions. The findings show that an alternative treatment method for cervical cancer can be developed with the application of CUR and DOX. Alternative methods for cervical cancer treatment may be developed using different methods in future studies.
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    Modulation of FOXP3 Gene Expression in OVCAR3 Cells Following Rosmarinic Acid and Doxorubicin Exposure
    (Mdpi, 2024) Toprak, Veysel; Ozdemir, Ilhan; Ozturk, Samil; Yanar, Orhan; Kizildemir, Yusuf Ziya; Tuncer, Mehmet Cudi
    Background/Objectives: Ovarian cancer has the highest mortality rate in the world. Treatment methods are listed as surgery, chemotherapy, and radiotherapy, depending on the stage of cancer, but developing resistance to chemotherapy increases the need for alternative agents that act on the same pathways. The effects of rosmarinic acid (RA) and doxorubicin (DX) on the activation of FOXP3, an important tumor suppressor gene, in OVCAR3 cells were examined. Materials and Methods: In this study, a human ovarian adenocarcinoma cell line was used. MTT analysis was performed to reveal the result of RA and DX on ovarian cancer cell proliferation. Expression levels of FOXP3 for cell proliferation and Capase-3 for apoptosis were determined by RT-qPCR. The wound healing model was applied to determine cell migration rates. The results were evaluated with one-way ANOVA in an SPSS 20.0 program as p <= 0.05. Results: It was determined that RA and DX alone and in combination inhibited the proliferation of OVCAR3 cells in different doses for 24, 48, and 72 h, and caused the cells to die by causing them to undergo apoptosis. Caspase-3 expression increased approximately tenfold in OVCAR3 cells, while FOXP3 expression was upregulated only in RA treatment and was downregulated in DX and RA + DX treatments. Conclusions: According to the results of our study, it was determined that the FOXP3 signaling pathway related to apoptosis, and proliferation was affected by the combination treatment of RA and DX in the OVCAR3 cancer cell line. This shows that RA will gain an important place in cancer treatment with more comprehensive study.
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    Thymoquinone Enhances Doxorubicin Efficacy via RAS/RAF Pathway Modulation in Ovarian Adenocarcinoma
    (MDPI, 2025) Toprak, Veysel; Ozdemir, Ilhan; Ozturk, Samil; Yanar, Orhan; Kizildemir, Yusuf Ziya; Tuncer, Mehmet Cudi
    Background/Objectives: Ovarian cancer remains one of the most commonly diagnosed malignancies among women worldwide. The heterogeneity among tumor subtypes and the emergence of treatment resistance have raised significant concerns regarding the long-term efficacy of chemotherapy, radiotherapy, and immunotherapy. In response to these challenges, drug repurposing strategies-utilizing existing drugs in novel therapeutic contexts-have gained increasing attention. This study aimed to investigate the cytotoxic and apoptotic effects of the combined application of doxorubicin (DX) and thymoquinone (TQ) on ovarian adenocarcinoma cells (OVCAR3). Methods: OVCAR3 cells were cultured in RPMI medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. Cell viability and proliferation were assessed using the MTT assay following treatment with various concentrations of DX and TQ. NucBlue immunofluorescence staining was employed to examine nuclear morphology and to identify apoptosis-associated changes. Additionally, quantitative real-time polymerase chain reaction (qRT-PCR) was per-formed to evaluate the expression levels of apoptosis-related and oncogenic pathway genes, including RAF, RAS, Bcl-2, and Bax. Results: The results demonstrated that the combination of DX and TQ significantly reduced OVCAR3 cell viability and induced apoptosis in a dose-dependent manner. qRT-PCR analysis revealed a downregulation of RAS, RAF, and Bcl-2 expression, along with an upregulation of Bax, indicating activation of the intrinsic apoptotic pathway. These findings suggest that thymoquinone exerts an-ti-proliferative and pro-apoptotic effects by modulating the RAS/RAF signaling cascade. Furthermore, the co-administration of thymoquinone with doxorubicin potentiated these effects, suggesting a synergistic interaction between the two agents. Conclusions: Histopathological and molecular evaluations further confirmed the activation of apoptosis and the suppression of key oncogenic pathways. Collectively, these results underscore the therapeutic potential of thymoquinone as both a monotherapy and an adjuvant to conventional chemotherapy, warranting further validation in preclinical and clinical studies.