Figure 1.
Chemical structure and antiproliferative activity against CCRF-CEM cells of acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a).
(A) Chemical structure of 8a. (B) Viability of CCRF-CEM cells after 24 h of exposure to 8a. The viability of the control cells, which were exposed to DMSO only, was set as 100%. n = 6, *p<0.05 compared with vehicle control. (C) Flow cytometric analysis of phosphatidylserine externalization (annexin V-binding) and cell membrane integrity (PI staining). CCRF-CEM cells were treated with 8a at 0.5 µM for 24 h.
Figure 2.
PCA scores plot resulting from the UPLC/MS spectra of CCRF-CEM cells with corresponding OPLS-DA for tested groups.
(A) PCA scores plot from control and 8a-treated group. (B) OPLS-DA scores plot from control and 8a-treated group. (C) Potential biomarkers in the S-plot between control and 8a-treated group.
Figure 3.
The comparison of biomarker intensity between control and compounds-treated cells.
These biomarkers involved in (A) Glutathione metabolism (B) Glycerophospholipid metabolism (n = 8), *p<0.05 and **p<0.01 compared with vehicle control.
Figure 4.
ROS and MDA levels in CCRF-CEM cells.
(A) The level of ROS after treatment with compound 8a (0–2 µM) for 24 h. (B) MDA production after 8a (0–2 µM) treatment for 24 h. Data represent the mean±SD, n = 3, *p<0.05 compared with vehicle control.
Figure 5.
Effects of 8a on the level of MMP and expression of apoptosis-related proteins.
(A) MMP affected by 8a in CCRF-CEM cells (n = 3). *p<0.05 compared with vehicle control. (B) Western blot analysis of the effect of 8a on cytochrome C release in CCRF-CEM cells treated with indicated doses of 8a for 24 h. (C) Representative Western blots for the expression of cleaved caspase-3 in CCRF-CEM cells following exposure to different concentrations of 8a for 24 h. β-actin (∼42 kDa) was used as a loading control.
Figure 6.
The hypothetic scheme of the action mechanism of 8a. Mito, mitochondrial.