K2 VİTAMİNİNİN OSTEOSARKOMA HÜCRELERİNDE ANTİPROLİFERATİF ETKİLERİ: FARKLI SİTOTOKSİSİTE ANALİZLERİNİN KARŞILAŞTIRILMASI

Abstract

Amaç Bu çalışmanın amacı, K2 vitamininin antiproliferatif etkilerini saptamak için yaygın olarak kullanılan dört sitotoksisite analizinin (XTT, nötral kırmızı alım, kristal viyole ve propidyum iyodür boyama) doğruluk potansiyelini karşılaştırmaktır. Gereç ve Yöntem Osteosarkoma hücre hattı olan Saos-2 hücreleri, 48 ve 72 saat boyunca K2 vitaminine (10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 100 μM ve 200 μM) maruz bırakıldı. Tetrazolyum tuzu testi (XTT), nötral kırmızı alım testi (NR), kristal viyole testi ve propidyum iyodür (PI) boyama yapılarak hücrelerin canlılık oranlarına göre K2 vitamini sitotoksisitesi belirlendi ve farklı dozların IC50 değerleri hesaplandı. Analiz sonuçları tek yönlü varyans analizi (ANOVA) ve Tukey testiyle değerlendirildi. Bulgular K2 vitamininin osteosarkoma hücreleri üzerinde sitotoksik etkileri olduğu sırasıyla XTT, NR, kristal viyole testi ve PI yöntemleri kullanılarak belirlendi ve IC50 değerleri 48 saatlik maruziyet için sırasıyla 61,93; 40,21; 62,11; 70,57 μM ve 72 saatlik maruziyet için sırasıyla 75,44; 68,22; 41,66; 88,01 μM olarak saptandı. Sonuç İstatistiksel analizler çalışmada uygulanan testlerden elde edilen sonuçların farklılık gösterdiğini ortaya koymuştur. Ayrıca propidyum iyodür boyama sonucundaki canlılık oranlarının diğer testlerle elde edilen sonuçlara kıyasla çarpıcı bir şekilde daha yüksek olduğu belirlenmiştir. Sitotoksisite analizlerinde sonuçların yanlış yorumlanmasını önlemek için farklı zamanlarda ve farklı konsantrasyonlarda inkübasyonlar gerektiği ve ayrıca tetrazolyum tuzu temelli testlerin metabolik olmayan diğer testlerle desteklenmesi gerektiği sonucuna varılmıştır.

Keywords

• İn vitro sitotoksisite
• Hücre canlılık testleri
• Osteosarkoma
• K vitamini.

ANTIPROLIFERATIVE EFFECTS OF VITAMIN K2 IN OSTEOSARCOMA CELLS: COMPARISON OF DIFFERENT CYTOTOXICITY ANALYZES

Abstract

Objective The aim of this study was to compare four commonly used cytotoxicity assays (XTT, neutral red uptake, crys- tal violet assay, and propidium iodide staining) which analyzed the antiproliferative effects of vitamin K2. Material and Method Saos-2 cells, an osteosarcoma cell line, were exposed to vitamin K2 at different concentrations (10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 100 μM and 200 μM) for 48 and 72 hours. Tetrazolium salt test (XTT), neutral red uptake (NR) assay, crystal violet assay, and propidium iodide (PI) staining were performed to determine cytotoxic potential of vitamin K2 in terms of the cell viability and IC50 values. The results were evaluated with one-way analysis of variance (ANOVA) and the Tukey test. Results Cytotoxic effects of vitamin K2 on osteosarcoma cells were analyzed with XTT, neutral red, crystal violet assay, and propidium iodide, respectively. IC50 values were determined exposure to 61.93; 40.21; 62.11; 70.57 μM vitamin K2 for 48 and 75.44; 68.22; 41.66; 88.01 μM vitamin K2 for 72 hours. Conclusion Statistical analysis revealed that there is a significant statistical difference between four tests used in this study. In addition, it was determined that the viability rates in propidium iodide staining were higher than other tests for cytotoxicity analyses. It has been concluded that incubations at different concentrations were required to prevent misinterpretation of results in cytotoxicity analyses, and tetrazolium salt-based tests should be supplemented with other non-metabolic tests.

Keywords

• In vitro cytotoxicity
• Cell viability tests
• Osteosarcoma
• Vitamin K.

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