Sümeyye AYDOGAN TÜRKOĞLU, Sinem GÜLTEKİN TOSUN, Feray KÖÇKAR

Farklı Hücre Hatlarında Hipoksik Koşullarda ADAMTS-2 İfadesinin Değişimi

ADAMTS’ler (A Disintegrin And Metalloproteaz With Trombospondin Motifs; Trombospondin Motifli Matriks Metalloproteazlar) dokuların yeniden düzenlenmesi, matriks yapımı ve yıkımı gibi fizyolojik olaylarda rol aldığı gibi kanser, kas-iskelet sistemi hastalıkları, enflamasyon, fibrozis gibi patolojik olaylarda da yer alır. ADAMTS ailesine ait 19 üye tanımlanmış ve birçok özelliğine göre gruplandırılmıştır. Bu gruplardan ADAMTS2, 3 ve 14 kollajen ADAMTS’ leri oluşturur ve kollajen işlenmesinde görev alır. ADAMTS2’nin önemli özelliklerinden biri kollajenlerin amino uçlarının kesilip uzaklaştırılmasında görev alması yeni tespit edilen diğer bir fonksiyonu ise anti-anjiyogenik aktiviteye sahip olmasıdır. Hipoksik regülasyon hücresel düzeyde çok sayıda genin ifadesini etkileyen bir süreçtir. Hücrelerde meydana gelen hipoksiya ile başta HIF-1 olmak üzere birçok gen aktive olur. HIF-1 hücresel oksijen konsantrasyonundaki değişimleri algılayarak yanıt oluşturmadan sorumlu transkripsiyon faktörüdür. Çalışmamızda ilk kez ADAMTS2 geninin hipoksik regülasyonu çalışılmış ve farklı hücre hatlarında hipoksik koşullardaki değişimi mRNA seviyesinde tespit edilmiştir. Çalışmamız kapsamında Saos-2, MG-63 (insan kemik karsinomu), PC-3, DU-145 (insan prostat karsinomu), MCF-7 (insan meme karsinomu), HT29, Colo-205 (insan kolon karsinomu), MKN-45 (insan gastrik karsinomu), Panc-1, Mia PaCa-2 (insan pankreas karsinomu), K-562 (insan kronik myeloid lösemi hücresi) ve HUVEC (insan umblikal ven endotel hücresi) hücre hatları seçilmiş ve yapılan analizlere göre ADAMTS2’ nin en çok MG-63 kemik karsinomda ifade olduğu tespit edilmiştir. Mia PaCa-2 ve K-562 hücrelerinde ise tespit edilebilir düzeyde ekspre olmadığı belirlenmiştir. Kimyasal hipoksi modeli oluşturulmuş ve ADAMTS2 mRNA seviyesi farklı hücre hatlarında incelenmiştir. DU-145, PC-3, HT-29, MCF-7 ve Saos-2 hücre hatlarında mRNA düzeyinde ADAMTS2 seviyesinin hipoksiyada arttığı tespit edilmiştir. En yüksek hipoksik cevap Saos-2 ve MCF-7 hücrelerinde gözlenmiştir.

Anahtar Kelimeler:

ADAMTS-2, Kollajen, Hipoksi, Anjiyogenez, Hücre Hattı

Variation of ADAMTS-2 Gene Expression in Hypoxic Conditions in Different Cell Lines

ADAMTS (A Disintegrin And Metalloproteinase with Trombospondin Motifs) is involved in pathological events such as cancer, fibrosis, musculoskeletal disorders, inflamation as well as physiological events such as tissue reorganization, matrix building and destruction. 19 members of the ADAMTS family have been identified and grouped according to many characteristics. The important features of ADAMTS2 is processing the amino ends of the collagen and also has an anti-angiogenic activity. Hypoxic regulation is a process that affects the expression of many genes at the cellular level. Hypoxia activates many genes, mainly HIF-1 transcription factor which is responsible for detecting changes in cellular oxygen concentration and for generating responses. For the first time, the hypoxic regulation of ADAMTS2 gene was studied and the variation in hypoxic conditions was determined at the mRNA level using different cell lines. We evaluated the effects of hypoxia in the expression level of ADAMTS-2 in different cell lines namely; Saos-2, MG-63 (bone carcinoma), PC-3, DU-145 (prostate carcinoma), MCF-7 (breast carcinoma), HT29, Colo-205 (colon carcinoma), MKN-45 (gastric carcinoma), Panc-1, Mia PaCa-2 (pancreatic carcinoma), K-562 (chronic myeloid leukemia cell) and HUVEC (umbilical vein endothelial cell). We found the most expression level in MG-63. Mia PaCa-2 and K-562 cells were not expressed detectable level of ADAMTS-2. A chemical hypoxic model was constructed and the level of ADAMTS2 mRNA was examined. It has been determined that ADAMTS2 mRNA level is increased in hypoxia in DU-145, PC-3, HT-29, MCF-7 and Saos-2 cells. The highest hypoxic response was observed in Saos-2 and MCF-7 cells.

Keywords:

ADAMTS-2, Collagene, Hypoxia, Angiogenesis, Cell Line,

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