Interfering with Color Response by Porphyrin-Related Compounds in the MTT Tetrazolium-Based Colorimetric Assay.

Autor: Choi BH; Division of Applied Food System, College of Natural Science, Seoul Women's University, 621 Hwarangro, Nowon-Gu, Seoul 01797, Republic of Korea., Kim MR; Division of Applied Food System, College of Natural Science, Seoul Women's University, 621 Hwarangro, Nowon-Gu, Seoul 01797, Republic of Korea., Jung YN; Division of Applied Food System, College of Natural Science, Seoul Women's University, 621 Hwarangro, Nowon-Gu, Seoul 01797, Republic of Korea., Kang S; Division of Applied Food System, College of Natural Science, Seoul Women's University, 621 Hwarangro, Nowon-Gu, Seoul 01797, Republic of Korea., Hong J; Division of Applied Food System, College of Natural Science, Seoul Women's University, 621 Hwarangro, Nowon-Gu, Seoul 01797, Republic of Korea.
Jazyk: angličtina
Zdroj: International journal of molecular sciences [Int J Mol Sci] 2022 Dec 29; Vol. 24 (1). Date of Electronic Publication: 2022 Dec 29.
DOI: 10.3390/ijms24010562
Abstrakt: Porphyrin compounds are widely distributed in various natural products and biological systems. In this study, effects of porphyrin-related compounds including zinc protoporphyrin (ZnPP), protoporphyrin IX (PPIX), cyanocobalamin (CBL), hemin, and zinc phthalocyanine (ZnPC) were analyzed on color response of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tetrazolium-based assay, a commonly-used method for analyzing cell viability. Color responses of MTT formazan formed in cells treated with ZnPP, PPIX, or ZnPC were significantly reduced even at submicromolar concentrations without affecting cell viability, whereas hemin and CBL did not. ZnPP, PPIX, and ZnPC rapidly induced degradation of MTT formazan already-produced by cells when exposed to light, but not under a dark condition. Photosensitizing properties of the three compounds were also verified through extensive generation of reactive oxygen species under light. The porphyrins did not affect the stability of water-soluble formazans including XTT, WST-1, WST-8, and MTS formazans. Several factors including different light sources and antioxidants modulated the degradation process of MTT formazan by the porphyrins. The results suggest that certain porphyrin compounds could cause a severe artifact in the MTT assay through rapid degradation of formazan dye due to their photosensitizing property, which needs to be considered carefully in the related assays.
Databáze: MEDLINE
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