Functional Analysis of Taube Nuss Gene in the Regulation of Cardiomyoblast Cells

Abstract

Differential gene expression is important in the regulation and maintenance of developmental stages and tissue specific physiological processes. The subtractive hybridization method is an efficient method for the detection of genes that have tissue specific expressions. In our previous work, our team had constructed a BALB/c mice heart tissue specific subtractive hybridization cDNA library. Taube Nuss (Tbn), which is a crucial transcription factor, is one of the genes that were isolated from this library. In this study, our aim was to analyze the expression of possible Tbn related genes and identify the Tbn pathway. In the scope of this study, the Tbn gene expression in BALB/c mice heart and skeletal tissue was analyzed with the Northern Blot technique. Then, the Tbn gene was silenced in H9c2 rat cardiomyoblast cell line using siRNA specific to the TBN gene. The gene expression levels of Pparg, Cebpa, Taf10 and Myocd genes, which were possibly related to the Tbn pathway, were analyzed in silenced cells. We observed that the expression of the Myocd gene was decreased, whereas the expression of the Pparg gene was increased in these cells. These results suggest that the Tbn gene plays a regulatory role in cardiac tissue. 

Keywords

• siRNA,H9c2,Tbn,Pparg,Taf10

Kardiyomiyoblast Hücre Regülasyonunda Taube Nuss Geninin Fonksiyonel Analizi

Abstract

Gelişim evrelerinin veya dokuya özgü fizyolojik süreçlerin düzenlenmesinde ve yürütülmesinde farklılaşmış gen ekspresyonu büyük önem taşımaktadır. “Subtractive” hibridizasyon yöntemi dokuya özgü ekspresyonu olan genlerin tespit edilmesi için oldukça etkin bir yöntemdir. Daha önceki çalışmalarda laboratuvarımızda BALB/c ırkı farelerde kalbe özgü ekspresyonu olan genleri içeren “Subtractive” hibridizasyon cDNA kütüphanesi oluşturulmuştur. Önemli bir transkripsiyon faktörü olan Taube Nuss (Tbn), bu kütüphaneden elde edilen genlerden birisidir. Bu çalışmadaki amacımız, Tbn geni ile ilişkili olduğu düşünülen diğer genlerin ekspresyonlarının araştırılması ve Tbn yolağının belirlenmesidir. Çalışma kapsamında BALB/c ırkı farelerden elde edilen kalp ve iskelet kası dokularına özgü Tbn geninin ekspresyonu Northern Blot analizi ile araştırılmıştır. Ardından, Tbn genine özgü siRNA’lar kullanılarak H9c2 rat kardiyomiyoblast hücre soyunda Tbn geni sessizleştirilmiştir. Yolak analizleri sonucunda Tbn ile ilişkili olabileceği tespit edilen Pparg, Cebpa, Taf10 ve Myocd genlerinin sessizleştirilen hücrelerdeki ekspresyon değişimleri araştırılmıştır. Bu hücrelerde, Myocd gen ekspresyonunun azaldığı ve Pparg gen ekspresyonunun arttığı saptanmıştır. Bu sonuçlar, Tbn geninin kalp dokusunda düzenleyici bir rol oynadığını işaret etmektedir. 

Keywords

• siRNA,H9c2,Tbn,Pparg,Taf10

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