Journal: PLoS ONE

Article Title: Rhizoma Paridis Saponins Suppresses Tumor Growth in a Rat Model of N -Nitrosomethylbenzylamine-Induced Esophageal Cancer by Inhibiting Cyclooxygenases-2 Pathway

doi: 10.1371/journal.pone.0131560

Figure Lengend Snippet: A . RPS inhibited the viability of esophageal cancer cells. After overnight incubation, cells in 96-well plates were treated with RPS at 0, 2.5, 5, 7.5, 10, 15, 20, 30, 40, or 60 μg/mL for 48 hours and then incubated with 20 μL of MTT (5mg/mL) for 4 hours. The absorbance at 550 nm was measured in a microplate reader. The percentage of viability relative to the controls without RPS was calculated. Each concentration was tested with 6 repeats in each experiment, n = 3. B . RPS reduced the invasion of esophageal cancer cells. Cells were cultured in the transwell chamber (5 x 10 4 /chamber, 8 μm pore size and coated with 1mg/mL matrigel) containing serum-free media with RPS at 0, 5, 10, or 20 μg/mL in the upper chamber and RPMI-1640 medium + 10% FCS in the lower chambers for 48 h. The penetrated cells at the lower surface of the filter were fixed and counted under a microscope. A total of 5 fields of each chamber were randomly selected and the average cell number of the 5 fields was used, n = 3. * and # represents significant difference at 5, 10, and 20 μg/mL of RPS compared to 0 μg/mL of RPS in EC9706 cell and KYSE150 cells, respectively, P < 0.01. C . Images of wound healing of EC9706 cells, 20 x. D . Images of wound healing of KYSE150 cells, 20 x. E . RPS reduced the migration of EC9706 cells. F . RPS reduced the migration of KYSE150 cells. A total of 5 scratches were used for each concentration of RPS. Cells were cultured in serum-free media with RPS at 0, 3.25, 7.5, or 15 μg/mL for EC9706 cells, or at 0, 5, 10, or 20 μg/mL for KYSE150 for 24 h. Wounds were photographed under a phase-contrast inverted microscope, and the percentage gap closure was calculated as (width at 0h –width at 24 n)/width at 0h * 100%, n = 3. * represents significant difference between RPS at specified concentration vs 0 μg/mL, P < 0.01.

Article Snippet: The absorbance at 550 nm was measured in a microplate reader (Corning Costar, Corning, NY, USA).

Techniques: Incubation, Concentration Assay, Cell Culture, Pore Size, Microscopy, Migration, Inverted Microscopy