Synthesis of new hetero-arylidene-9(10H)-anthrone derivatives and their biological evaluation

In Vitro and In Vivo Effect of Palladacycles: Targeting A2780 Ovarian Carcinoma Cells and Modulation of Angiogenesis

“…Cell suspensions were stained with 0.4% trypan blue and placed on a hemocytometer (Hirschmann, Eberstadt, Germany) and counted on an Olympus BX51 microscope (Olympus, Tokyo, Japan). 33,34 Evaluation of the Mitochondrial Membrane Potential. 5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolocarbocyanine iodide (JC-1; Abnova Corp., Walnut, CA) was used to determine the mitochondrial membrane potential as previously described.…”

“…5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolocarbocyanine iodide (JC-1; Abnova Corp., Walnut, CA) was used to determine the mitochondrial membrane potential as previously described. 34 Briefly, 2 × 10 5 cells/mL from the A2780 cell line were seeded in 6-well plates and, after 24 h, challenged with the IC 50 concentrations of compounds 3 and 9, as described elsewhere, for 48 h. 34 Cells were then stained with a JC-1 solution according to the manufacturer's instructions. Green and red JC-1 fluorescences were quantified with the BL-1 and BL-2 filters of the Attune acoustic focusing cytometer (Thermo Fisher Scientific), and the information was treated with the Attune Cytometric Software 2.1.…”

“…Green and red JC-1 fluorescences were quantified with the BL-1 and BL-2 filters of the Attune acoustic focusing cytometer (Thermo Fisher Scientific), and the information was treated with the Attune Cytometric Software 2.1. 34 ROS Production in A2780 Cells. As previously described, A2780 cells plated in 6-well plates at a density of 2 × 10 5 cells/well were treated, after an initial incubation of 24 h, with media with IC 50 concentrations of compounds 3 and 9, 25 μM H 2 O 2 (positive control), and 0.1% DMSO (vehicle control) and incubated for 48 h. 30,32,34 Cells were then treated with TripLE Express (Thermo Fisher Scientific), washed twice with PBS (1×), and then resuspended in 10 μM 2,7-dichlorodihydrofluorescein diacetate in PBS (1×).…”

“…34 ROS Production in A2780 Cells. As previously described, A2780 cells plated in 6-well plates at a density of 2 × 10 5 cells/well were treated, after an initial incubation of 24 h, with media with IC 50 concentrations of compounds 3 and 9, 25 μM H 2 O 2 (positive control), and 0.1% DMSO (vehicle control) and incubated for 48 h. 30,32,34 Cells were then treated with TripLE Express (Thermo Fisher Scientific), washed twice with PBS (1×), and then resuspended in 10 μM 2,7-dichlorodihydrofluorescein diacetate in PBS (1×). Cells were then incubated for 30 min at 37 °C.…”

“…Cells were then incubated for 30 min at 37 °C. 30,32,34 This compound will become fluorescent after the removal of acetate groups by intracellular esterases in the presence of ROS. 8,10,12 Attune Cytometric Software 2.1 was used to record data obtained from cells analyzed in the Attune acoustic focusing cytometer (Thermo Fisher Scientific).…”