Değişik Glukoz Konsantrasyonlarının E2F1 Seviyesi ve Hücre Proliferasyonuna Etkisi

Öz

Amaç: E2F1 transkripsiyon faktorü, hücre döngüsünün kontrolü, proliferasyon, hücre farklılaşması, DNA tamiri ve apoptosis üzerinde düzenleyici etkisine ek olarak enerji metabolizmasının düzenlenmesinde ve glukoz kullanımında da anahtar rol oynayan bir transkripsiyon faktorüdür. Bu nedenle çalışmamızda, farklı glukoz konsantrasyonlarının E2F1 protein ekspresyonuna ve bunun hücre proliferasyonuna etkisinin olup olmadığını belirlemeye çalıştık. Gereç ve Yöntemler: Çalışmamızda İnsan Embriyonik Böbrek Hücre Hattı olan HEK293 hücrelerinde farklı glukoz konsantrasyonlarının, E2F1 ekspresyonuna olan etkisi Western Blot SDSPage yöntemi ile ve hücre içerisinde sentezlenen E2F1 protein miktarını artırmak için kullanılan ektopik E2F1 ekspresyonunun hücre proliferasyonuna olan etkisi MTT yöntemi ile gösterilmiştir. Bulgular: Farklı glukoz konsantrasyonlarının E2F1 protein seviyelerini etkilediği tespit edilmiştir. HEK293 hücrelerinde 2 g/L glukoz konsantrasyonunda en yüksek proliferasyon gözlenirken, 20 g/L glukoz konsantrasyonunda negatif bir etki gözlenmiştir. E2F1 transfekte hücrelerinde farklı glukoz konsantrasyonlarının hücre proliferasyonuna anlamlı bir etkisi saptanmamıştır.Sonuç: Artan glukoz konsantrasyonu E2F1 protein seviyesinde artışa yol açmıştır. 20 g/L değerinde gözlenen negatif etki glukoz toksisitesi ile uyumludur

Anahtar Kelimeler

• E2F1, Enerji metabolizması, Glukoz, Hücre proliferasyonu, Western Blot, MTT

The Effect of Different Glucose Concentrations on E2F1 Level and Cell Proliferation

Abstract

Objective: The E2F1 transcription factor plays a key role in the control of the cell cycle, proliferation, cell differentiation, DNA repair, and apoptosis as well as regulation of energy metabolism and glucose utilization. Therefore, we wanted to determine whether different concentrations of glucose would affect the cellular level of E2F1 and whether this would also affect cellular proliferation.Material and Methods: In this study, the effect of different glucose concentrations on E2F1 expression in the Human Embryonic Kidney Cell Line which is called HEK293 cells was shown by the Western Blot SDS-Page method and the effect of ectopic E2F1 expression, which was used to increase the amount of E2F1 protein synthesized in the cell, on cell proliferation was shown by the MTT method.Results: We detected that different glucose concentrations affected E2F1 levels differently. The highest proliferation was observed at the 2 g/L glucose concentration in the HEK293 cells, while a negative effect was detected with a 20 g/L glucose concentration. E2F1 transfected cells did not show any significant effect on cell proliferation of different glucose concentrations.Conclusion: Increased glucose concentration led to an increase in the E2F1 protein level. The observed negative effect at 20 g/L is compatible with glucose toxicity

Keywords

• E2F1, Energy metabolism, Glucose, Cell proliferation, Western Blot, MTT

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