Siklofosfamid, Saos-2 Osteosarkom Hücrelerinde MMP-2 ve -9'un Ekspresyonunu Azaltarak Anti-Metastatik Etki Gösterir

Amaç: Osteosarkoma, ergenler ve genç yetişkinler arasında en sık rastlanan primer malign kemik tümörü olmakla birlikte, kanser gelişiminin altında yatan moleküler mekanizmalar tam anlamıyla aydınlatılamamıştır. Siklofosfamid (CYC), çeşitli kanserlerin ve kronik inflamatuvar hastalıkların tedavi rejimlerinde oldukça yaygın kullanılan alkilleyici antineoplastik bir ajandır. Bu çalışmanın amacı, CYC’nin Saos-2 hücreleri üzerine olan etkisini kanser progresyonu ilişkili genlere ait bazı matriks metalloproteaz (MMP-2 ve MMP-9) proteinlerinin ekspresyon seviyelerini belirleyerek moleküler düzeyde ortaya koymaktır. Araçlar ve Yöntem: Geliştirilen insan osteosarkoma (Saos-2) hücre kültürlerinde CYC uygulanarak sitotoksisite analizleri yapıldı. Belirlenen dozda CYC, iki boyutlu hücre hatlarına 12, 24 ve 48 saatlik sürelerde uygulandı. Ajan uygulaması sonucunda bu kültürlerdeki metastatik belirteçler olan MMP’lerin ekspresyon seviyeleri immünositokimyasal yöntem ile belirlendi. Bulgular: MMP2 ve MMP-9 protein ekspresyon seviyeleri oluşturulan hücre kültürlerinde artan süre ile korele olarak kontrol (PBS) gruplarında artarken; CYC uygulaması ile birlikte azalmıştır. Sonuç: Saos-2 hücrelerine CYC uygulaması ile MMP-2 ve MMP-9 gibi metastazda rol oynayan genlerin ekspresyon düzeylerini azaltıldığı belirlendi. Bu sonuçlar osteosarkoma kanseri tedavisi için moleküler çalışmalara ışık tutacak niteliktedir.

Anahtar Kelimeler:

kanser, siklofosfamid, MMP, osteosarkoma, Saos-2

Cyclophosphamide Exerts an Anti-Metastatic Effect by Reducing the Expression of MMP-2 and -9 in Saos-2 Osteosarcoma Cells

Purpose: Although osteosarcoma is the most common primary malignant bone tumor among adolescents and young adults, the molecular mechanisms underlying the development of cancer are not fully elucidated. Cyclophosphamide (CYC) is an alkylating antineoplastic agent widely used in the treatment regimens of various cancers and chronic inflammatory diseases. The aim of this study is to reveal the effect of CYC on Saos-2 cells at the molecular level by determining the expression levels of some matrix metalloprotease proteins (MMP-2 and MMP-9) of cancer progression-related genes. Materials and Methods: Cytotoxicity analyzes were performed by applying CYC to the developed human osteosarcoma (Saos-2) cell cultures. The determined dose of CYC was applied to the 2D cell lines for 12, 24 and 48 hours. As a result of agent application, the expression levels of MMPs, which are metastatic markers in these cultures, were determined by the immunocytochemical method. Results: While MMP-2 and MMP-9 protein expression levels increased in control (PBS) groups in correlation with the extent of duration in the cell cultures created, decreased with CYC administration. Conclusion: It was determined that the expression levels of genes that play a role in metastasis, such as MMP-2 and MMP-9, were decreased by CYC application to Saos-2 cells. These results shed light on molecular studies for the treatment of osteosarcoma cancer.

Keywords:

cancer, cyclophosphamide, MMP, osteosarcoma, Saos-2,

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