A.Tuğşen AYDEMİR, Meltem ALPER, Feray KÖÇKAR

ADAMTS-3’ ün (A Disintegrin and Matrix Metalloproteinase Type,1 Motif 3) Normal ve Kanserli Hücre Hatlarındaki Ekspresyon Paterni

ADAMTS-3, temel görevi kıkırdak ve kemiğin temel kollajeni olan prokollajen II’ yi kesmek olan bir prokollajen amino proteinazdır. Bu olgunlaştırma işlemi, dokular için doğru fibril konformasyonunun oluşmasını sağlar. ADAMTS-3, kollajen II’ nin yanı sıra kollajen III ve fibronektin gibi diğer hücreler arası matriks bileşenlerini de işeyebilmektedir. Hücreler arası matriks bileşenlerinin ADAMTS-3 aracılı işlenmesi hem normal hem de patolojik durumlarda dikkate değer bir öneme sahiptir. Bu yüzden aile üyelerinin substrat spesifikliği ve dokulardaki ekspresyon profilleri ilgi çekici bir konu haline gelmiştir. Hatta aile üyelerinden bazılarının, bazı tümör tiplerinde ekspresyonlarının değiştiği aydınlatılmıştır. Bu yüzden protümör ve tümör supresör fonksiyonları araştırılmaktadır. ADAMTS-3’ün ekspresyon profili sadece birkaç normal ve kanserli dokuda çalışılmıştır. ADAMTS-3’ün hücre hatlarındaki ekspresyon paterni hakkındaki bilgi oldukça kısıtlıdır. Bu çalışmada normal olarak HUVEC ve kanserli hücre hatlarından, Saos-2, MG-63, DU-145, PC-3, Hep3B ve PANC-1 hücrelerinde ADAMTS-3 mRNA (sqRT-PCR) ve protein ekspresyon düzeyleri (western-blot) belirlenmiştir.HUVEC, Saos-2, MG-63 ve PC-3 hücrelerinde ADAMTS-3’ün yüksek düzeyde ifade olduğu belirlenmiştir. Bu bulgular, ADAMTS-3’ün bazı spesifik kanser türlerindeki transkripsiyonel regülasyonu ve ADAMTS-3’ün tümör gelişimine ve metastazına olan katkılarının belirlenmesi üzerine yapılacak çalışmalar için temel olacaktır.

Anahtar Kelimeler:

ADAMTS-3, gen ekspresyonu, kanser hücre hatları, sqRT-PCR, western-blot

Expression Pattern of ADAMTS-3 (A Disintegrin and Matrix Metalloproteinase Type,1 Motif 3) in Normal and Cancer Cell Lines

ADAMTS-3 is a procollagen N- proteinase whose main function is to cleave procollagen II, the principal collagen of the cartilage and bone. This maturation process provides accurate fibril conformation for tissues. Along with collagen II, ADAMTS-3 can also process some other extracellular matrix components such as collagen III and fibronectin. ADAMTS mediated processing of the extracellular matrix components has considerable importance in both normal and pathological circumstances. Therefore, substrate specificities and the tissue expression profiles of the family members have become a hot topic. Further, altered expressions of the family members have been elucidated in some tumor types. Therefore, protumor and /or tumor suppressor functions of them is being investigated. The expression profile of the ADAMTS-3 has been studied only in a few normal and cancer tissues. Knowledge of the expression patterns of the ADAMTS-3 in cell lines are rather limited. In the present study, we determined ADAMTS-3 mRNA (sqRT-PCR) and protein expression levels (western-blot) in normal HUVEC, and cancer cell lines, namely, Saos-2, MG-63, DU-145, PC-3, Hep3B and PANC-1 cells. ADAMTS-3 was highly expressed in HUVEC, Saos-2, MG-63, and PC-3 cells. These findings will be fundamental for future studies that will be performed on ADAMTS-3 transcriptional regulation studies specific cancer types and also defining of the ADAMTS-3 contribution on the development and metastasis of specific cancer types.

Keywords:

ADAMTS-3, gene expression, cancer cell lines, sqRT-PCR, western-blot,

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