i.e., cell invasion, migration, and colony formation were studied
using established protocols. The Boyden chamber method with
slight modifications was used to study the inhibitory effects of EOs on
the invasion of HCT 116 cells. A total of 50 μL of Matrigel (5 mg/mL,
diluted with RMPI 1:1) was loaded into each well of a 96-well plate
and allowed to polymerize for 45 min at 37 °C in an incubator. A
total of 5 × 103 cells in 150 μL of RPMI medium containing 25, 50,
and 90 μg/mL of EOs (IC25, IC50, and IC90) were added to each well
separately and incubated for 24 h. Cells with 0.5% DMSO in media
and 5-FU (5 μg/mL) were used as the negative and positive control,
respectively. At the end of the incubation period, non-invading
cellswerewashedwith PBS and the number of invading cellswas determined
under inverted lightmicroscopy. The results were presented
as the mean percentage inhibition compared to untreated cells
i.e., cell invasion, migration, and colony formation were studiedusing established protocols. The Boyden chamber method withslight modifications was used to study the inhibitory effects of EOs onthe invasion of HCT 116 cells. A total of 50 μL of Matrigel (5 mg/mL,diluted with RMPI 1:1) was loaded into each well of a 96-well plateand allowed to polymerize for 45 min at 37 °C in an incubator. Atotal of 5 × 103 cells in 150 μL of RPMI medium containing 25, 50,and 90 μg/mL of EOs (IC25, IC50, and IC90) were added to each wellseparately and incubated for 24 h. Cells with 0.5% DMSO in mediaand 5-FU (5 μg/mL) were used as the negative and positive control,respectively. At the end of the incubation period, non-invadingcellswerewashedwith PBS and the number of invading cellswas determinedunder inverted lightmicroscopy. The results were presentedas the mean percentage inhibition compared to untreated cells
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