In Vitro and In Vivo Anticancer Effects of Sterol Fraction from Red Algae Porphyra dentata
Figure 7
In vivo effect of P. dentata F2 fraction (a) and in vitro effect of F2 fraction and various agents (b) on ROS production by MDSCs. In vivo, tumor-bearing BALB/c mice were i.p. injected with F2 fraction at dosages of 5 and 25 mg/kg body weight. MDSCs were isolated from tumor-bearing control mice and F2-treated mice as described in Section 2 and their ROS productions were measured. In vitro, MDSCs isolated from the tumor-bearing control mice were incubated first with F2 fraction or the tested agent; thereafter, ROS production was measured. Where the stimulating agent PMA was used, MDSCs were incubated at 37°C for 5 min with 1 μg/mL PMA and then washed with cold PBS. To block ROS production in vitro, MDSCs were incubated with catalase, arginase inhibitor (nor-NOHA), or F2 fraction at 37°C for 10 min. The oxidative-sensitive dye DCFDA was used to measure ROS production by the cells. MDSCs were incubated with 2 μM DCFDA for 15 min at 37°C and then washed twice with PBS. The intensity of fluorescence was measured by flow cytometry. Data were obtained from one experiment, with 4 individual mice from each group. Measurements were obtained in duplicate and the mean fluorescence intensity (MFI) was calculated for MDSCs in each group, for both in vivo and in vitro treatments.
