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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 Inhibition of voltage?gated potassium channels affect expressions of miR-126 and miR-126* in breast cancer cell lines(AEPress, 2020) Turgut Coşan, Didem; Soyocak, Ahu; Öner, ÇağrıAIM: We aimed to determine the possible correlation between voltage?gated potassium channels and micro RNAs in breast cancer and metastatic breast cancer cells. METHOD: Kv1.3 and Kv10.1 channels were inhibited by specific siRNAs using a lipofectamine-based transfection in MCF-7 and MDA-MB-231 cells. After transfection, total RNA was isolated, and then miR-126 and miR-126* expressions were observed using RT-PCR. RESULTS: There was a negative correlation between Kv channels and miRNAs according to the characteristics of the breast cancer cells. The inhibition was observed not only in Kv1.3 but also in Kv10.1 in MCF-7 cells, and miR-126 and miR-126* expressions were downregulated compared to the control group (p < 0.001). The inhibition of these channels in MDA-MB-231 cells caused an upregulation of miR-126 and miR-126* expressions (p < 0.001). CONCLUSION: The miR-126 and miR-126* expressions differed according to benign and malign breast cancer cell lines. Furthermore, we found that miR-126/126* may interact with Kv1.3 and Kv10.1 voltage-gated potassium channels. Our study suggests and indicates the relationship between Kv channels and miRNAs in breast cancer cells (Tab. 1, Fig. 2, Ref. 51).Yayın Meme kanserinde Kv 1.3 ve Kv 10.1 voltaj bağımlı potasyum kanallarının inhibisyonunun oksidatif stres üzerindeki rolü(Dicle Üniversitesi, 2017) Turgut Coşan, Didem; Öner, Çağrı; Soyocak, Ahu; Metcalfe, Evrim; Djamgoz, MustafaAmaç: Reaktif oksijen türevleri oksidatif stres, iyonize radyasyon maruziyeti, iskemi-reperfüzyon hasarı ve kanseri içeren fizyolojik ve patolojik durumlarda artmaktadır. Çalışmamızda meme kanser hücrelerinin farklılaşmasında ve çoğalmasında etkili olduğu düşünülen voltaj kapılı potasyum kanallarının inhibisyonunun etkilerini gözlemek amaçlandı. Yöntemler: Farklı karakterlerdeki meme kanseri hücrelerine bu potasyum kanallarına özgü siRNA’ların transfeksiyon işlemi yapıldı. İnkübasyon sonrasında hücre lizatları hazırlanarak antioksidan ve oksidan seviyeleri belirlendi. Elde edilen verilerle oksidatif stres hesaplandı. Sürekli değişkenlerin normal dağılıma uygunluğu Kolmogorov-Smirnov testi kullanılarak yapıldı. Normal dağılım gösteren değişkenlerin gruplar arasındaki karşılaştırmaları tek yönlü varyans analizi ile değerlendirildi. Çoklu karşılaştırmalar ise Tukey HSD testi ile gerçekleştirildi. Bulgular: Kanal inhibisyonu ile invaziv olmayan karakterdeki MCF-7 hücrelerinin oksidatif stres seviyesinde düşüş olduğu gözlendi. İnvaziv karakterdeki MDA-MB-231 hücrelerinde ise Kv 1.3 siRNA transfekte edilen grupta oksidatif stres seviyesinde düşüş belirlenirken, Kv 10.1 siRNA transfekte edilen grupta artış belirlendi. Sonuç: Voltaj kapılı potasyum iyon kanallarının inhibisyonunun kanser hücrelerinde oksidatif stres oluşturabileceği belirlenmiştir. Ayrıca, çalışmamızdaki en önemli bulgu kanser hücrelerinde voltaj bağımlı potasyum kanallarının oksidatif stres üzerinde etki yapabileceği ve bunun hücrelerin metastatik karakteri ve iyon kanalı türü ile bağlantılı olmasıdır.Yayın The silenced potassium channels Kv1.3 and Kv10.1 affects miR-126 in breast cancer cells(Nature, 2018) Turgut Coşan, Didem; Soyocak, Ahu; Öner, ÇağrıIntroduction: It has been shown that non-coding regulatory RNAs, especially play a role in formation and progression of cancer. miR-126 is markedly downregulated in human breast cancer tissues. Various studies indicate that some voltage gated potassium channels are also overexpressed in the tumor. Kv1.3, a member of the voltage-gated potassium channel family is overexpressed in cancerous breast tissues than in normal breast tissues. Another protein, Kv10.1 is overexpressed in many human tumors, including breast cancer and it has oncogenic properties. Ether à go-go 1 (Eag1) channel is overexpressed in a variety of cancers and plays important roles in cancer progression. However, the association between with voltage gated potassium channels and miR-126 is unclear. In our study, we aimed to show whether there is association between miR-126/miR-126* and Kv1.3 or Kv10.1 in non-invasive estrogen positive MCF-7 and invasive estrogen negative MDA-MB-231 human BC cells. Materials and Methods: MCF-7 and MDA-MB-231 cells were transfected with Kv1.3 and Kv10.1 specific siRNA. miR-126 and miR-126* expression was determined in total RNA isolated from the cells. All samples were normalized to the internal controls, and fold changes were calculated through relative quantification (2???Ct). Results: The results of our research indicate that there is a strong inverse relationship with the expression levels of miR-126 and potassium channels. Conclusions: The miR-126/126* expressions increased in the MDA-MB-231 cells and decreased in the MCF-7 cells by using siRNA against potassium channels Kv1.3 and Kv10.1.Yayın Sphingosine-1-phosphate (S1P) receptors in nonmetastatic and metastatic breast cancer patients(Nature, 2018) Turgut Coşan, Didem; Soyocak, Ahu; Çalış, I. U.; Öner, Çağrı; Metcalfe, E.; Erkasap, S.; Mutlu, F.Introduction: S1P receptors S1P1 and S1P3 encourage cancer progression. In this study, the aim was to evaluate the functional roles in the progression of disease by determining the expression of S1P1 and S1P3 in non-metastatic and metastatic breast cancer. Materials and Methods: Breast tissue specimens were obtained by surgery and reduction mammoplasty. The peripheral mononuclear cells (PBMC) of non-metastatic, metastatic breast cancer patients and noncancerous individuals were also taken. Total RNA isolation was carried out by tissues and cells. Gene expression of S1P1 and S1P3 was determined by Real-Time PCR. Results: In this study, S1P1 and S1P3 gene expressions in tissues (1.03 and 1.16 fold, respectively) were found to be higher than PBMC samples in non-metastatic breast cancer patients. In addition, in PBMC samples S1P1 levels decreased by 2.90 fold in patients with non-metastatic breast cancer, increased by 1.33 fold in patients with metastatic breast cancer. S1P3 levels decreased by 3.35 fold in patients with non-metastatic breast cancer and 1.45 fold in patients with metastatic breast cancer. Conclusions: The result of our study showed that both gene expressions were higher in cancerous tissues than in normal tissues. The expression of the S1P1 gene in PBMCs is increased in nonmetastatic breast cancer patients compared to non-cancer individuals, while it is increased in metastatic breast cancer patients. S1P3 is lower in both non-metastatic and metastatic individuals than non-cancer individuals. These results indicate that, for the S1P receptors in the studies done in PBMC and tissue samples should be considered different expression properties.
