The importance of protein expression SOD2 in response to oxidative stress for different cancer cells

DNA hasarı, onarım mekanizmasındaki bozukluklar ve kanser arasındaki nedensel ilişki deneysel ve epidemiyolojik veriler ile gösterilmiştir. DNA hasarının oksidatif strese dönüşüm aşamaları baskılayıcı genlerin mutasyonal inaktivasyonu ve onkogenlerin aktivasyonu ile ilişkilendirilir. Bu amaç doğrultusunda, kanser hücrelerinin SOD2 (Superoksit dismutaz 2) değişimine bağlı olarak oksidatif stres koşullarındaki etkileri ve bu mekanizmada NF-kB (Nükleer Faktör kappa B) transkripsiyon faktörünün önemi gösterilmiştir. Hücre hatları ATCC’nin (Amerikan tipi kültür kolleksiyonu) belirttiği prosedürlere uygun olarak kültüre edilerek saklandı. Hücreler gruplara ayrıldıktan sonra, MNNG ve tempol ajanlarının optimal konsantrasyonun hücre canlılık (MTT test) yöntemiyle belirlenmesiyle belirtilen zaman aralıklarında uygulandı. Hücre hatlarındaki farklı SOD2 protein düzeyleri western blot analizleri ile gösterilerek karşılaştırıldı. Bununla birlikte transfeksiyon yoluyla yapılan luciferase yöntemi kullanılarak NF-kB enzimatik aktivitesi belirlendi. Bunun sonucunda, 20 μM MNNG uygulandığında yüksek NF-kB aktivitesi gözlenirken, 30 μM tempol tedavisi ile NF-kB seviyesinin azaldığı görüldü. Sonuçlar SOD2 ekspresyonu ve NF-kB aktivasyonu arasında güçlü korelasyon gösterdi.

SOD2 Salınımının farklı kanser türlerinde oksidatif strese karşı değişimi ve önemi

As a causal relationship between DNA damage, repair mechanism disorders and cancer demonstrated by experimental and epidemiological data. Transforming process starts from DNA damage to oxidative stress is associated with mutational inactivation of suppressor genes, activation of oncogenes. Related to this aim, it is important to indicate how cancer cells react under oxidative stress through by SOD2 expression, besides presenting correlation between NF-kB mechanism. Cell lines were maintained and cultured as recommended by ATCC (American type culture collection) resource. Cells were grouped and after detecting optimum concentration by MTT test (cell viability assay), treated with DNA damage agent (MNNG) and antioxidant (tempol) for indicated time points. Western blot analysis revealed that cell lines with comparable levels of SOD2 (Superokside dismutase 2) protein expression. However, cells were collected to measure NF-kB (Nuclear Factor kappa B) enzymatic activity using luciferase expression by transfection way. We observed high constitutive NF-kB activity by using 20 μM MNNG although decreasing NF-kB during 30 μM tempol treatment. The results showed strong correlation between SOD2 expression and NF-kB activation.

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