Osteosarkom Hücre Hattında Hedefe Spesifik RANKL İnhibitörü Olarak Gambojik Asidin Antikanser Profili

Year 2021, , 442 - 452, 31.08.2021

Aykut Özgür , Esra Duman

https://doi.org/10.18185/erzifbed.928668

Abstract

Osteosarkom, genç nüfus arasında yaygın bir kanser türüdür ve genellikle büyüyen kemiklerde gelişir. Osteosarkom vakalarının yaklaşık yüzde yirmisi metastaz eğilimi gösterir ve osteosarkomlu hastaların tedaviden sonra hayatta kalma oranı düşüktür. Kemiğin yeniden şekillenmesinin RANK / RANKL / OPG anahtar düzenleyici üçlüsü, osteosarkomun tümör oluşumunda kritik rol oynar. Gambojik asidin anti-proliferatif aktivitesi, XTT testi ile Saos-2 hücre hattında belirlendi. Gambojik asidin antikanser aktivitelerini anlamak için RANK-RANKL kompleksi ile moleküler docking hesaplamaları da yapıldı. RANKL ve OPG'nin ekspresyon seviyeleri, RT-PCR ve ELISA deneyleri ile gen ve protein seviyesinde ölçüldü. Osteosarkom hücrelerine karşı in vitro potansiyel anti-invazif özelliği, yara iyileştirme deneyi kullanılarak değerlendirildi. Deneysel analizler, gambojik asidin Saos-2 hücrelerinde hücre proliferasyonunu, hücre göçünü ve RANKL / OPG'nin protein ekspresyon oranını azalttığını gösterdi. Gambogic asit, 549.38 nM'lik Kd değeri ve -8.54 kcal / mol'lük tahmini serbest enerji ile RANK-RANKL kompleksine bağlandı. Gambojik asidin, hedefe özgü osteosarkom tedavisi için önemli ilaç şablonu olduğu bulunmuştur.

Keywords

Osteosarkom, Saos-2, Gambojik asit, RANK, RANKL, OPG

Supporting Institution

Tokat Gaziosmanpaşa University

Project Number

2019/86

Anticancer Profiling of Gambogic Acid as a Target Specific RANKL Inhibitor in Osteosarcoma Cell Line

Abstract

Osteosarcoma is a common cancer type among the youth population and usually develops in growing bones. Approximately twenty percent of osteosarcoma cases show tendency to metastases and patients with osteosarcoma have a low survival rate after treatment. RANK/RANKL/OPG key regulator triad of bone remodeling play critical roles in tumourigenesis of osteosarcoma. Anti-proliferative activity of the gambogic acid was determined in Saos-2 cell line by XTT assay. To understand the anticancer activities of the gambogic acid, molecular docking calculations were also performed with RANK-RANKL complex. The expression levels of the RANKL and OPG was measured at gene and protein level with RT-PCR and ELISA assays. Its potential anti-invasive property in-vitro against osteosarcoma cells was evaluated using wound healing assay. Experimental assays indicated that gambogic acid suppressed cell proliferation, cell migration, and decreased protein expression ratio of RANKL/OPG in Saos-2 cells. Gambogic acid binds to RANK-RANKL complex with Kd value of 549.38 nM and with estimated free energy of binding -8.54 kcal/mol. Gambogic acid is found to be significant drug template for target specific osteosarcoma treatment.

Keywords

Osteosarcoma, Saos-2, Gambogic acid, RANK, RANKL, OPG

Project Number

2019/86

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