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Yayın 4-hydroxycoumarin effects on both cellular and genetic characteristics of hepatocellular carcinoma cells(Springerlink, 2022) Öner, Çağrı; Soyergin, Dilara; Özyurt, Ahmet; Çolak, Ertuğrul4-Hydroxycoumarin is an aromatic substance which is metabolized in liver and used as a therapeutic agent for various diseases. We aimed to determine the impact of 4-Hydroxycoumarin on HepG2 cells according to their viability, proliferation, adhesion and gene expression of cellular behavior parameters. Inhibitory concentration 50 (IC50) of 4-Hydroxycoumarin was detected on HepG2 cells. After determining the optimal time and concentration, the effect of 4-Hydroxycoumarin on viability, proliferation and adhesion of HepG2 cells were observed. Gene expressions of Ki-67, MMP-2, MMP-9 and piR-823 expression were determined by using Real Time-Polymerase Chain Reaction. IC50 value of 4-Hydroxycoumarin on HepG2 cells was 5 ?M at the 48 h (p < 0.001). 5 ?M at the 48 h of 4-Hydroxycoumarin caused to decrease of proliferation (p < 0.001) and viability of HepG2 cells (p < 0.001). Viability rate were supported by hematoxylin-eosin staining. Adhesion of cells increased on 4-Hydroxycoumarin treated cells compared to control (p < 0.001). While Ki-67 gene expression of 4-Hydroxycoumarin treated group decreased (p < 0.001); upregulation of MMP-2, MMP-9 and piR-823 expressions were determined in 4-Hydroxycoumarin treated group (p < 0.001). According to the cellular and genetic perspective, 4-Hydroxycoumarin might be useful to treat hepatocellular carcinoma. High adhesion and proliferation are the main characteristics of HepG2 cells, 4-Hydroxycoumarin treatment caused to lose these functions. The genetic markers of these characteristics also supported the same result. These are first findings about the effect of 4-Hydroxycoumarin on piR-823 and genes which are key features of cellular survival mechanisms.Yayın Different approaches for breast cancer: voltage gated potassium channels and micrornas(Ankara University Faculty of Sciences, 2015) Öner, Çağrı; Turgut Coşan, Didem; Çolak, ErtuğrulMicro RNAs and voltage-gated potassium channels (VGPCs) both play critical roles in the development of cancer. We aimed to reveal the diversity of miR-126/126*, which effects angiogenesis and vascular development through the inhibition of VGPCs. In this study, potassium channel inhibitors, including tetraethylammonium (5mM), 4-aminopyridine (5mM), margatoxin (1µM) and astemizole (2µM), were applied to MCF-7 and MDA-MB-231 breast cancer cell lines. After totally isolating RNA from the cells, Real-Time Polymerase Chain Reaction was used in order to identify gene expressions. OneWay ANOVA was used for variation analyses, while Tukey HSD and Tamhane were used to assess whether multiple comparisons were statistically significant (P < 0.001). Our results showed an increase in miR-126/126* expressions after the channel inhibition of MCF-7 and MDA-MB-231 cell lines (P < 0.001). miR-126/126* expressions were increased using TEA, 4-AP and astemizole in both cell lines. miR-126/126* expressions were only increased through the use of margatoxin in MCF-7. miR-126/126* may interact with voltage-gated potassium channels. In our study, the inhibition of K channels using K channel blockers resulted in an increase of miR-126/126* expression. Therefore, our data suggested that there could be another perspective between K channels and non-coding RNAs in the development of breast cancer.Yayın Estrogen and androgen hormone levels modulate the expression of PIWI interacting rna in prostate and breast cancer(Public Library of Science, 2016) Öner, Çağrı; Turgut Coşan, Didem; Çolak, ErtuğrulPIWI interacting RNAs (piRNAs), a member of non-coding RNA, originate from intergenic repetitive regions of the genome. piRNA expressions increase in various cancers and it is thought that this increase could be caused by hormones. We aimed to determine the effects of hormones on piRNA expression in breast and prostate cancer. High viability and a decrease in adhesion were observed at the concentrations of the highest proliferation. Furthermore, an increase in adhesion was also observed in MDA-MB-231 cells. After hormone treatment, while piR-651 expression had increased both breast and prostate cancer cell lines, piR-823 expressions increased in prostate cancer cell lines and only in the breast cancer cell line which was malignant. Thus, it was determined that piR-823 might show different expressions in different type of cancers.Yayın Evaluation of paraoxonase-1 enzyme activity and oxidative stress relations in malignant mesothelioma cases(Turkish Respiratory Society, 2020) Turgut Coşan, Didem; Ak, Güntülü; Çolak, Emine; Dal, Aylin; Öner, Çağrı; Soyocak, Ahu; Çolak, Ertuğrul; Güneş, Hasan Veysi; Metintaş, MuzafferBACKGROUND: Malignant pleural mesothelioma (MPM) is the most common cancer in the pleura and highly aggressive with a very poor prognosis. Asbestos, known as a carcinogenic mineral with fiber structures, is the main cause of MPM formation. Exposure to asbestos causes an increase in reactive oxygen species, deficiency of antioxidant enzyme levels, and DNA damage. As a result of asbestos pathogenesis, all of these changes cause pulmonary fibrosis, pleural diseases, and malignancies. The endogenous antioxidant paraoxonase-1 (PON-1) is a calcium-dependent esterase involved in the hydrolysis of lipid peroxides, and PON-1 has been shown to have protective properties in oxidative stress and inflammatory diseases in various studies. OBJECTIVE: The study aimed to examine the relationship of MPM with PON-1 enzyme activity and oxidative status using total oxidant status (TOS) and total antioxidant status (TAS). MATERIALS AND METHODS: The study population was formed of 33 retrospectively examined mesothelioma patients as MPM group and 33 age- and sex-matched healthy individuals as controls. PON-1 activity was measured spectrophotometrically by enzyme-linked immunosorbent assay method. Total antioxidant and oxidant status was determined using Rel Assay Diagnostics kit. Oxidative stress index (OSI) was estimated as the ratio of the TOS to the TAS levels. RESULTS: In the present study, PON-1, TOS, TAS, and OSI levels were adjusted by comorbidity and smoking. The results indicated that TOS and OSI of MPM patients increased compared to healthy controls (P < 0.001 for both). The results also demonstrated the decrease of PON-1 activity and TAS in MPM cases (P < 0.001, for both). CONCLUSION: These results suggested that oxidative stress occurring as a result of inhalation of asbestos fibers may reduce the level of PON-1.Yayın Genetic markers indicate that 1,25-dihydroxyvitamin d treatment may not protect against hepatocellular carcinoma(Kare Publishing, 2021) Öner, Çağrı; Çolak, ErtuğrulObjectives: The impact of 1,25-dihydroxyvitamin D on hepatocellular carcinoma (HCC) cells is a complicated area. In this study, we aimed to clarify the effect of 1,25-dihydroxyvitamin D on HCC cells according to genetic markers. Methods: The optimal concentration of 1,25-dihydroxyvitamin D is treated to HepG2 cells (250 nM at the 48th hour). From treated HepG2 cells, total Ribonucleic Acid (RNA) was isolated, and Ki-67, MMP-2, MMP-9, HIF-1?, hTERT, and piR823 gene expressions were determined by SYBR Green-based real-time polymerase chain reaction. Results: : Increased expressions of Ki-67, hTERT, and piR-823 were determined compared with the control group at the 48th hour after treatment (p<0.001), while decreased gene expressions of MMP-2, MMP-9, and HIF-1? were observed compared with the control group (p<0.001). Conclusion: Currently, there are several different opinions about the usage of vitamin D, especially in HCC. In addition to researchers who argue that vitamin D has anticarcinogenic and protective properties, an increasing number of researchers argue that tumor cells can become aggressive after HCC occurs. According to our results, it was determined that vitamin D causes HepG2 HCC cells to become aggressive in terms of gene expression in the parameters used as a marker for proliferation, adhesion, and differentiation.Yayın Hepatoselüler karsinom tedavisi için etkili terapötikler PIWI Interacting RNA ifadelerini nasıl değiştirir?(Alanya Alaaddin Keykubat Üniversitesi, 2020) Öner, Çağrı; Altıner, Necdet; Çolak, ErtuğrulAmaç: PIWI interacting RNA’lar (piRNA’lar) herhangi bir protein üretemeyen ancak hücrelerin transkripsiyon ve transkripsiyon sonrası mekanizmalarında etkili olan küçük kodlayıcı olmayan RNA'ların yeni üyeleridir. Günümüzde, kanser hücrelerinin tedavisinde kimyasal bileşikler yerine, hem doğal bileşikler hem de vitaminler uygulanabilirliği araştırılmaktadır. Bu çalışmadaki amacımız, 4-Hidroksikoumarinin ve aktif D vitamini formunun (1.25-dihidroksivitamin D) hepatoselüler karsinomada piRNA'ların olası ekspresyonları üzerindeki değişiklikleri belirlemektir.Yöntemler: Önceki çalışmamızdan elde edilen verilere göre, optimal zaman ve konsantrasyonu belirlenen 4-Hidroksikoumarin, 1.25-dihidroksivitamin D ve D vitamininin ilaç formu HePG2 hücrelerine uygulandı. Uygulamadan sonra total RNA izole edildi. piR-Hep-1 ve piR-651'in ekspresyonları Gerçek Zamanlı Polimeraz Zincir Reaksiyonları kullanılarak belirlendi.Bulgular: Elde edilen verilere göre, 4-Hidroksikoumarin uygulanan HePG2 hücrelerinde kontrole göre piR-651 ekspresyonunda istatistiksel olarak anlamlı bir artış gözlenmiştir (p< 0.001). Bununla birlikte, 4-Hidroksikoumarin uygulamasından sonra piR-Hep-1 ekspresyonundaki değişim istatistiksel olarak anlamlı değildir (p> 0.05). Buna karşılık, 1,25- dihidroksivitamin uygulaması HePG2 hücrelerinde piR-Hep-1 ekspresyonunu istatistiksel olarak anlamlı şekilde azaltmıştır (p <0.001). D vitamininin ilaç formunun uygulamasından sonra piR-Hep-1 ekspresyonundaki azalma istatistiksel olarak anlamlı değildir (p> 0.05).Sonuç: Tüm bu veriler, piRNA'ların bazılarının hepatoselüler karsinomda özel ekspresyon paternlerine sahip olabileceğini ve bu ekspresyon paternlerinin, uygulanan doğal bileşikler tarafından düzenlenebileceğini göstermektedir. Hepatoselüler karsinomda tek tek etkili olduğu gözlenen maddelerin, beklentilerin aksine farklı piRNA ekspresyon değişikliklerine neden olabileceğini savunmaktayız.Yayın PIWI interacting rna-651 inhibition transforms the genetic features of mcf-7 breast cancer cells(Tech Science Press, 2021) Öner, Çağrı; Çolak, ErtuğrulpiRNAs are novel members of small non-coding RNAs and have an impact on genetic and epigenetic mechanisms of cells. It was aimed to investigate the role of piR-651 on MCF-7 benign breast cancer cells by focusing on molecular characteristics. Anti-piR-651 was transfected and effects of piR-651 on proliferation, adhesion, and motility of MCF-7 cells were detected after the 24th, 48th, and 72nd hour. Gene expressions of piR-651, Ki-67, MMP-2, ER?, HIF-1?, and hTERT were determined by using RT-PCR. piR-651 inhibition caused the decrease of proliferation, adhesion (p < 0.001), and motility of MCF-7 cells. The efficiency of anti-piR-651 transfection supported the determination of the decrease of piR-651 expression after transfection after the 48th hour (p < 0.001). AntipiR-651 transfection caused the downregulation of Ki-67, MMP-2, ER?, HIF-1?, and hTERT gene expressions after the 48th hour (p < 0.001). In MCF-7 cells, piR-651 inhibition can change both cellular characteristics and gene expressions which were related to these characteristics. piR-651 inhibition causes the decrease of both proliferation and adhesion of cells, which are especially important cellular marks of MCF-7 cells. These results have shed light on whether we can mitigate the effects of cancer cells via piRNA inhibition. The absence of piRNA expression caused the change of fate of benign breast cancer cells by decreasing malign features of MCF-7 cells.Yayın PIWI interacting rna-823: epigenetic regulator of the triple negative breast cancer cells proliferation(Kare Publishing, 2022) Öner, Çağrı; Çolak, ErtuğrulObjectives: Triple negative breast cancer cells are estrogen, progesterone, and HER receptor negative, malign breast cancer cells. These cells have high telomerase activity, and this activity gives these cells high proliferation and evading apoptosis abilities. In this study, we aimed to determine the affect piR-823 on genetic parameters of proliferation and ER? status in MDA-MB-231 cells. Methods: After anti-piR-823 transfection, proliferation of cells was tested by XTT. Gene expressions were determined by RT-PCR. Protein expressions were determined by ELISA. Results: The proliferation decreased after inhibition (p<0.001). Gene and protein expressions of ER? were upregulated while hTERT was downregulated after inhibition (p<0.001). Furthermore, piR-823 inhibition cause to decrease PI3K/ AKT/mTOR gene expressions and miR-126 expression (p<0.001). Conclusion: Obtained data indicates that piR-823 inhibition lead MDA-MB-231 cells to increase ER? expression. Decreased expressions of hTERT and PI3K/AKT/mTOR pathway affect cell proliferation. Moreover, miR-126 decrease indicates that piRNAs and miRNAs can share the same target molecules and piRNA expression changes might have an impact on miRNA expressions. All obtained data is important on the perspective of piRNAs and their effect on cellular characteristics of triple negative breast cancer cells.Yayın Vitamin D’nin hepatoselüler karsinom üzerindeki etkisi(Eskişehir Osmangazi Üniversitesi, 2020) Öner, Çağrı; İsan, Hatice; Aktaş, Ranan Gülhan; Çolak, ErtuğrulHepatoselüler karsinoma (HCC) dünya çapında görülme sıklığı açısından en çok görülen dördüncü kanser türüdür. Bu kanser türünün görülme nedenlerinin başında Tip II diyabet, obezite ve alkol kullanımı gelmektedir. Çalışmamızda, karaciğer hastalıklarında etkili olan, ancak hücresel mekanizmalar açısından etkileşimi henüz tam olarak belirlenemeyen vitamin D’nin hem ilaç formu hem de aktif formu olan 1,25-dihidroksivitaminin [1,25(OH)2D3] HepG2 hepatoselüler karsinom hücrelerinin karakteristik özelliklerinde oluşturduğu değişimlerin belirlenmesi amaçlanmıştır. HepG2 hücrelerine vitamin D’nin hem ilaç formu hem de 1,25(OH)2D3 formu ayrı ayrı uygulanarak en etkili konsantrasyonları ve saatleri belirlendi. Bu aşamadan sonra uygun konsantrasyon ve saatte uygulanan her iki maddenin HepG2 hücrelerinin proliferasyonu, adezyonu ve immunohistokimyasal olarak p53 miktarındaki etkileri belirlendi. Elde edilen verilere göre, 250 nM vitamin D’nin ilaç formu HepG2 hücrelerine uygulandıktan 96 saat; 250 nM 1,25(OH)2D3 uygulandıktan sonra 48 saat sonra kontrol grubuna göre istatistiksel olarak proliferasyonun en çok görüldüğü konsantrasyon ve saat olarak belirlendi. Hem vitamin D’nin ilaç hem de 1,25(OH)2D3 formu HepG2 hücrelerinin adezyonunu kontrol grubuna göre istatistiksel olarak anlamlı bir şekilde azaldığı belirlendi (p<0.001). Optimal konsantrasyon ve saatte uygulanan her iki vitamin D formunun p53 miktarındaki değişimleri immunohistokimyasal olarak incelendiğinde, kontrol ve sham gruplarına göre azaldığı gözlendi. Hepatoselüler kanser hastalarında uygulanacağı zaman, vitamin D dozuna dikkat edilmesi ve sürekli kontrol altında olunması yönünde ön veri sağlamaktadır. Her ne kadar bazı karaciğer hastalıklarından korunmak için önemli bir vitamin olmasına rağmen; kanser hücrelerinde proliferasyonu arttırması, adezyonu azaltması ve bir tümör baskılayıcı olan p53 miktarını azaltması konu üzerinde soru işaretleri yaratmaktadır. Vitamin D’nin hepatoselüler hastalar üzerinde uygulanması konusunda farklı bir bakış açısı yaratan sonuçlarımız, ilerleyen dönemlerde konu ile alakalı yapılacak olan çalışmalar önemli bir ön veri niteliği taşımaktadır.
