Metabolic activity of viable cells was measured by MTT assay based on the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium salt to a formazan product, which can be quantified spectrophotometrically to determine the percent of viable cells [32 (link)].
Anticancer activities of new benzothiazole acylhydrazones derivatives were evaluated against A549 C6, MCF-7 and HT-29 cell lines. The selectivity of their cytotoxic effects was evaluated on NIH/3T3 mouse embryonic fibroblast cells. NIH/3T3 cells were incubated in DMEM supplemented with fetal calf serum, penicillin (100 IU/mL), streptomycin (100 mg/mL) and 7.5% NaHCO3 at 37 °C in a humidified atmosphere of 95% air and 5% CO2. Carcinogenic cells were incubated in RPMI medium supplemented with fetal calf serum, penicillin (100 IU/mL), streptomycin (100 mg/mL) and 7.5% NaHCO3 at 37 °C in a humidified atmosphere of 95% air and 5% CO2. All cell lines were seeded at a density of 1 × 104 cells into the 96-well plates. After 24 h of incubating period, the culture mediums were removed and test compounds were added at concentrations of 0.000316–1 mM. After a 24 h incubation period, OD of samples were measured by a microplate reader (Biotek, Winooski, VT, USA) at 540 nm. Inhibition % at concentrations was determined using the formula below and IC50 values were calculated by nonlinear regression analysis using the SigmaPlot v.10 package program (Manufacturer, City, US State abbrev. if applicable, Country) [33 (link),34 (link),35 (link),36 (link)]. Cisplatin was used as a positive control:
Anticancer activities of new benzothiazole acylhydrazones derivatives were evaluated against A549 C6, MCF-7 and HT-29 cell lines. The selectivity of their cytotoxic effects was evaluated on NIH/3T3 mouse embryonic fibroblast cells. NIH/3T3 cells were incubated in DMEM supplemented with fetal calf serum, penicillin (100 IU/mL), streptomycin (100 mg/mL) and 7.5% NaHCO3 at 37 °C in a humidified atmosphere of 95% air and 5% CO2. Carcinogenic cells were incubated in RPMI medium supplemented with fetal calf serum, penicillin (100 IU/mL), streptomycin (100 mg/mL) and 7.5% NaHCO3 at 37 °C in a humidified atmosphere of 95% air and 5% CO2. All cell lines were seeded at a density of 1 × 104 cells into the 96-well plates. After 24 h of incubating period, the culture mediums were removed and test compounds were added at concentrations of 0.000316–1 mM. After a 24 h incubation period, OD of samples were measured by a microplate reader (Biotek, Winooski, VT, USA) at 540 nm. Inhibition % at concentrations was determined using the formula below and IC50 values were calculated by nonlinear regression analysis using the SigmaPlot v.10 package program (Manufacturer, City, US State abbrev. if applicable, Country) [33 (link),34 (link),35 (link),36 (link)]. Cisplatin was used as a positive control:
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