Fig 1.
Strategy for the design of dimers of EM-A derivatives 1a-2b.
Table 1.
In vitro inhibitory effects of compounds 1a-2b against the proliferation of three human cancer cell lines.
Fig 2.
Inhibitory and apoptosis effects of compound 1b on cell proliferation in Hela and MCF-7 cells.
(A) Inhibitory effects of compounds 1b. The cells were treated with various doses of 1b for 6, 12, 24, 36, 48, 72 h. The inhibitory ratio was measured by MTT assay. (B) Apoptosis effects of compound 1b. The cells were cultured with 0.5, 1.5, 2.5 μM 1b for 24 h, stained with PI at 4°C for 30 min, and measured by flow cytometry after collection. The percentage of cells in different phases of the cell cycle was represented by a bar diagram. All of the values were mean±SD of 3 separate experiments.
Table 2.
In vitro antiproliferative activities of compound 1b against six human cancer cell lines.
Fig 3.
Apoptosis and DNA fragmentation induced by compound 1b on HeLa and MCF-7 cells.
(A) Cell morphology observation under a phase contrast microscopy after incubation with medium or 1.5 μM of compound 1b for 24 h. (B) Cellular morphologic observation under a fluorescence microscopy by AO-EB staining. (C) DNA fragmentation observation. (Scale bar = 10 μm)
Fig 4.
Flowcytometric analysis of AnnexinV binding to the membrane phospholipid phosphatidylserine in Hela and MCF-7 cells.
After treatment with 0.5, 1.5, 2.5 μM of compound 1b for 24h, the cells were stained with AnnexinV-FITC and propidium iodine, and then subjected to the flow cytometric analysis.
Fig 5.
Mitochondrial potential alternation of HeLa and MCF-7 cells by compound 1b.
After treatment with 0.5, 1.5, 2.5 μM of 1b for 24h, the cells were stained with 5g/L Rhodamine 123. Fluorescent density reflected mitochondrial transmembrane potential was determined by flow cytometric analysis. Values were mean±SD of 3 separate experiments.
Fig 6.
Expression of Bax, Bcl-2, translocation of Cyt c and Capase-3 in HeLa and MCF-7 cells.
The cells were treated with different concentration of 1b (0, 0.5, 1.5 and 2.5 μM) for 24h.