In the first method, the dsDNA-GC surface was dried under a stream of nitrogen, after which the electrode was coated with 20 μL of a solution of QPhNO2 in ethanol P.A., allowed to rest for 5 min and then dried again under a stream of nitrogen until the gel was completely dry. After this step, 5 mL of acetate buffer was added to the cell, and DPV experiments were conducted. In the second method, the biosensor was immersed in a solution of QPhNO2 (5, 10 or 20 μM) for 15 min, after
which electrochemical measurements were taken immediately. The same procedure was also applied to the biosensor immersed only in acetate buffer. Single-stranded DNA (ssDNA) was prepared click here by dissolving 3.0 mg of dsDNA in 1.0 mL of chloridric acid (1 M) and heating for 1 h until complete dissolution. This treatment was followed by neutralizing the solution with 1.0 mL of sodium hydroxide (1 M) and adding 9 mL of acetate buffer (Diculescu et al., 2005). Freshly prepared ssDNA solution was added to the cell, and single-scan DPV experiments were conducted in the range check details of 0 to +1.4 V vs. AgAgCl, Cl− (0.1 M). Two peaks corresponding to the oxidation of the guanine and adenine bases appeared at potentials of +0.815 V and +1.131 V, respectively. After the first run, the
electrode was washed, polished and returned to the ssDNA solution. After cleaning the surface, the GC electrode was inserted into a solution containing QPhNO2 (at different concentrations of 5–46 μM) and ssDNA, and the DPV experiment was repeated. A clean GC electrode was also employed in the DPV experiments involving a 20 μM solution of QPhNO2 alone, and the current of peak Ia was used for comparison. The IC50 values for the MTT assay were obtained by nonlinear regression using the GRAPHPAD program (Intuitive Software for Science, San Diego, CA) from 3 to 4 independent
experiments performed in triplicate. The data are presented as the means ± S.D. from three independent experiments. Differences between experimental groups were compared by one-way ANOVA followed by Newman–Kells test for multiple comparison (p < 0.05), whereas Student’s t tests were used to compare data obtained in the absence or presence of NAC (p < 0.05). The inhibitory effects of nor-beta and its nitrophenylamine derivative QPhNO2 were initially determined Amino acid on the growth of HL-60 cells. The HL-60 cell line could be considered a suitable model to study compounds derived from beta-lapachone because the cytotoxic effects and apoptosis-inducing properties of this compound have already been demonstrated using this cell line (Planchon et al., 1995 and Planchon et al., 1999). As shown in Table 1, both QPhNO2 and nor-beta exhibited a strong inhibitory effect on HL-60 cell proliferation after 24 h of incubation, with IC50 values of 0.32 and 2.01 μM, respectively, while doxorubicin showed an IC50 value of 0.22 μM (Table 1).
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