Tetrazolium Bazlı Sitotoksisite Testleri Her Zaman Doğru Sonuçları Yansıtmayabilir

Abstract

Background: Cell viability and/or cytotoxicity analysis is one of the most important tools used for biological evaluation in vitro studies. The selection of the right cytotoxicity tests is critical to form the basis for in vivo and preclinical studies, specifically for cancer research. In the present study, we aimed to investigate the cytotoxic effects of bromelain, a widely-used phytochemical product in the medical field, and idarubicin, an anthracycline antibiotic used in the treatment of cancer, in normal lymphocytes and a promyelocytic leukemia cell line (HL-60) with MTT, WST-1, and luminescent ATP assays and to compare the results of these tests..Materials and Methods: We obtained peripheral blood lymphocytes from healthy, young, non-smoker male volunteers and obtained the HL-60 cell line from the American Type Culture Collection (ATCC). Bromelain and idarubicin were added in increasing concentrations to both cell lines. Cells were incubated at 37°C in a carbon dioxide incubator for 24 h. After incubation, cytotoxicity levels were determined by MTT, WST-1, and ATP assays, and morphological evaluations were performed by fluorescent staining.Results: The MTT and WST-1 assays demonstrated that cell viability/formazan formation increased with bromelain concentration; however, the luminescent ATP assay demonstrated that cell viability decreased with increasing concentrations of bromelain. Whereas fluorescent staining methods confirmed the ATP assay results, the MTT and WST-1 assays contradicted the ATP assay results. The cytotoxic effects of idarubicin were similar in the two cell lines according to the three different measurement methods and were positively correlated with the results of the fluorescent staining methods.Conclusion: The detection of cell viability and cytotoxicity by bromelain with the MTT and WST-1 assays in lymphocytes and HL-60 cells is limited. To obtain accurate and reliable results from cytotoxicity studies, a measurement method should be carefully selected by considering that the phytochemicals to be tested could interfere with the results, and the results should be verified by other methods.