Feray KÖÇKAR, Sümeyye AYDOGAN TÜRKOĞLU, Arife Sinem GÜLTEKİN

Kanser hücrelerinde Hipoksik Koşullarda ADAMTS2 İfadesinin Değişimi

Motifli Matriks Metalloproteazlar) dokuların yeniden düzenlenmesi, matriks yapımı ve yıkımı gibifizyolojik olaylarda rol aldığı gibi; kanser, kas-iskelet sistemi hastalıkları, enflamasyon, fibrozis gibipatolojik olaylarda da yer alır. ADAMTS ailesine ait 19 üye tanımlanmış ve birçok özelliğine göregruplandırılmıştır. Bu gruplardan ADAMTS2, 3 ve 14 kollajen ADAMTS’leri oluşturur ve kollajenişlenmesinde görev alır. ADAMTS2’nin önemli özelliklerinden biri kollajenlerin amino uçlarının kesilipuzaklaştırılmasında görev alması yeni tespit edilen diğer bir fonksiyonu ise anti-anjiyogenik aktiviteyesahip 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 hücreseloksijen konsantrasyonundaki değişimleri algılayarak yanıt oluşturmadan sorumlu transkripsiyonfaktörüdür. Çalışmamızda ilk kez ADAMTS2 geninin hipoksik regülasyonu çalışılmış ve farklı hücrehatlarında hipoksik koşullardaki değişimi mRNA seviyesinde tespit edilmiştir. Hipoksinin ADAMTS2 ifadedüzeyi üzerindeki etkileri Saos-2, MG-63 (kemik karsinomu), PC-3, DU-145 (prostat karsinomu), MCF-7(meme karsinomu), HT29, Colo-205 (kolon karsinomu), MKN-45 (gastrik karsinomu), Panc-1, Mia PaCa2(pankreas karsinomu), K-562 (insan kronik myeloid lösemi hücresi) ve HUVEC (insan umblikal venendotel hücresi) hücrelerinde değerlendirilmiştir. ADAMTS2’ nin en çok MG-63 kemik karsinomda ifadeolduğu tespit edilmiştir. Mia PaCa-2 ve K-562 hücrelerinde ise tespit edilebilir düzeyde ekspre olmadığıbelirlenmiştir. Kimyasal hipoksi modeli oluşturulmuş ve ADAMTS2 mRNA seviyesi farklı hücre hatlarındaincelenmiştir. DU-145, PC-3, HT-29, MCF-7 ve Saos-2 hücre hatlarında mRNA düzeyinde ADAMTS2seviyesinin hipoksiyada arttığı tespit edilmiştir. En yüksek hipoksik cevap Saos-2 ve MCF-7 hücrelerindegözlenmiştir.

Variation of ADAMTS2 Gene Expression in Hypoxic Conditions in Cancer Cells

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 ADAMTS2 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 ADAMTS2. 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.

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