Determination of early spectral changes in melanoma cells during epoxomicin-induced apoptotic process
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Tarih
2021
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Refik Saydam National Public Health Agency (RSNPHA)
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Objective: Determining the changes in the apoptosis process in cells can provide important information for new treatment and drug research. Apoptotic cells undergo a series of subcellular changes that lead to cell shrinkage and fragmentation. Determining these early changes in the apoptotic process depending on time may provide a new perspective to cell culture studies. The aim of this study is to develop a method in which early spectral changes occurring in the apoptosis process of melanoma cells can be determined depending on time. Methods: In this study, epoxomicin (Epo) was used to induce apoptosis in A375 melanoma cell line and apoptotic dose was determined by MTT method. In order to evaluate the early apoptotic process in a time-dependent manner, measurements were made at five different time points (0.5-6 hours) with a fixed Epo dose. DCFH-DA method was used to measure ROS, which is the most important stimulus of apoptosis. Bax amount was determined by Western Blot technique. Spectroscopic measurements were made with a back-reflection spectroscopy experiment setup consisting of spectrometer, tungsten-halogen light source and fiber optic probe. Apoptosis index values were determined from spectra. Results: Early spectral changes were determined with the spectra measured in the apoptosis time of melanoma cells. A significant difference was found in ROS measurements at 2, 4 and 6 hours compared to control. Cell viability was found to be 70% lower than control at 75 and 100 nM Epo doses after 24 hours. Time-dependent Bax levels were found to increase in all Epo groups as an indicator of apoptosis. Spectroscopic apoptosis index value was found to be compatible with ROS and Bax results at all groups. Conclusion: A new approach has been presented in which spectral changes occurring in the early stage of the apoptosis process in cell culture can be determined by back reflection spectroscopy. This approach has the potential to be developed in cell culture studies as a method that can monitor apoptosis over time without interfering with cell culture conditions. © 2021. All Rights Reserved.
Açıklama
Anahtar Kelimeler
apoptosis, apoptozis, Back reflection spectroscopy, epoksomisin, epoxomicin, Geri yansıma spektroskopisi, melanoma, melanoma
Kaynak
Turk Hijyen ve Deneysel Biyoloji Dergisi
WoS Q Değeri
Scopus Q Değeri
Q4
Cilt
78
Sayı
3
