Cytotoxicity evaluation of a Thai herb using tetrazolium (MTT) and sulforhodamine B (SRB) assays

Abstract

Background: Various assays are used to evaluate the cytotoxic effect of chemicals on cultured cells. The sulforhodamine B (SRB) colorimetric assay is based on the ability of the SRB dye to bind basic amino acid residues on proteins. In contrast, the MTT (dimethylthiazol-diphenyltetrazolium bromide) colorimetric assay is based on mitochondrial uptake and succinate dehydrogenase reduction of soluble, yellow, MTT tetrazolium salt to an insoluble blue MTT formazan product. The aim of this study was to evaluate the cytotoxicity of a Thai herb by comparing MTT and SRB assay results. Methods: Mouse fibroblast (L929) cells were exposed to 0.01, 0.1, 0.25, and 0.5% (w/v) of a Thai herb in a 96-cluster well culture plate for 24 h. Cell viability after exposure to the Thai herb was determined by MTT and SRB assays in separate tissue culture plates. The two assays were compared using intra-class correlation coefficient (ICC) analysis. Results: There were no significant differences between the two cytotoxicity assays (p > 0.05). The ICC values showing the agreement of the two assays in the negative and positive control groups and Thai herb concentrations of 0.01, 0.1, 0.25, and 0.5% were 0.93 and 0.99 and 0.53, 0.51, 0.95, and 0.98, respectively. Conclusions: In general, the MTT and SRB assays performed similarly, exhibiting moderate to excellent correlation in the evaluation of the cytotoxicity of a Thai herb.

Antitumor, cytotoxic, and antioxidant evaluation of six heterocyclic compounds containing different heterocycle moieties

“…In the SRB assay, the dye binds to basic amino acids of cellular proteins and cell density determination refers to total protein mass . While, tetrazolium (MTT) technique depended upon mitochondrial dehydrogenase enzyme in which these enzymes can reduce the tetrazolium dye to its insoluble formazan with a purple color …”

“…found that isonicotinic acid hydrazide, owing hydrazide moiety, cause human hepatoblastoma cell line (HepG2), healthy cell, injury by mitochondrial DNA damage . In addition to, MTT is based on mitochondrial uptake . From all the previous reasoning, it can be easily explained the difference between the obtained results from two assays.…”

“…The tetrazolium (MTT) test was the most commonly used in latest years . The colorimetric test for MTT (dimethylthiazol‐diphenyltetrazolium bromide) is based on mitochondrial uptake and succinate dehydrogenase reduction of soluble, yellow, MTT tetrazolium salt to an insoluble blue MTT formazan product . In contrast, colorimetric assay for the sulforhodamine B (SRB) is based on the ability of the SRB dye to bind basic amino acid residues on proteins …”

“…[6] The colorimetric test for MTT (dimethylthiazol-diphenyltetrazolium bromide) is based on mitochondrial uptake and succinate dehydrogenase reduction of soluble, yellow, MTT tetrazolium salt to an insoluble blue MTT formazan product. [7] In contrast, colorimetric assay for the sulforhodamine B (SRB) is based on the ability of the SRB dye to bind basic amino acid residues on proteins. [7] Several of the present treatments have problems with toxicity and drug-resistance, these encouraged researchers to explore effective new treatments for cancer disease.…”

“…[7] In contrast, colorimetric assay for the sulforhodamine B (SRB) is based on the ability of the SRB dye to bind basic amino acid residues on proteins. [7] Several of the present treatments have problems with toxicity and drug-resistance, these encouraged researchers to explore effective new treatments for cancer disease. [8] Consequently, synthesis of new bioactive compounds such as heterocyclic compounds is an important aim for many researchers.…”