Effect of combination of artesunate and arsenic trioxide on the proliferation and apoptosis of NB4 cells

Abstract

Objective To investigate the effects of artesunate combined with arsenic trioxide (ATO) on the proliferation and apoptosis of NB4 cells. Methods The NB4 cells were treated with different concentrations of artesunate and arsenic trioxide respectively for 48 h. The cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) method. The cells were divided into 4 groups: control group, artesunate group, arsenic trioxide group, and the combination of artesunate and arsenic trioxide group. The cell cycle and apoptosis were detected by flow cytometry (FCM). The protein expression levels of Bcl-2 and Bax were detected by Western blot. Results The MTT results showed that compared with the control group, the proliferation inhibition rates of 0.25, 0.50, 1.00, 2.00, 4.00 μmol/L artesunate group (19.26% ± 3.59%, 36.53% ± 2.67%, 61.32% ± 2.50%, 70.30% ± 3.15%, 86.92 ± 0.02%) significantly increased (P<0.05); the proliferation inhibition rates of 1, 2, 4, 8, 16 μmol/L arsenic trioxide group (12.69% ± 2.43%, 64.26% ± 2.02%, 85.10% ± 2.67%, 92.06% ± 2.21%, 93.67% ± 3.36%) significantly increased (P<0.05); and the proliferation inhibition rate (40.17% ± 5.49% vs. 32.23% ± 3.52%) of combination of artesunate and arsenic trioxide group significantly higher than the arsenic trioxide group (P<0.05). Compared with the arsenic trioxide group, the percentage of G0/G1 phase cells (74.20% ± 1.43% vs. 66.14% ± 1.78%), the apoptosis rate (58.00% ± 2.41% vs. 34.57% ± 1.22%), and the expression level of Bax protein (1.35 ± 0.09 vs. 1.13 ± 0.09) in the combination of artesunate and arsenic trioxide group significantly increased (P<0.05), the expression level of Bcl-2 protein (0.45 ± 0.09 vs. 1.03 ± 0.10) in the combination of artesunate and arsenic trioxide group significantly decreased (P<0.05). Conclusions Artesunate can significantly enhance the proliferation inhibition and apoptosis induced by arsenic trioxide on NB4 cells. The possible mechanism of proliferation inhibition and apoptosis of NB4 cells by artesunate combined with arsenic trioxide may be related to reduce the expression of anti-apoptotic protein Bcl-2 and increase the expression of apoptotic protein Bax. Key words: Cell proliferation; Apoptosis; Artesunate; Arsenic trioxide; Genes, bcl-2; bcl-2-Associated X protein; NB4 cells