Fig 1.
Treatment with DHA and DHA/HTF decreased Jurkat cell viability.
Jurkat cells (1 × 104/well) were exposed to DHA (0, 2.5, 5, 10, 20, 40, 80 μM) for 24, 48 or 72 h(a) or DHA/HTF for 48 h(b). The cell viability was assessed by CCK-8 assay. The results represented mean ± standard deviation. ***p<0.001, compared with the control group, # p<0.001, compared with DHA.
Fig 2.
HTF enhanced ROS production following DHA exposure.
Jurkat cells (1 × 106/well) were exposed to DHA (0, 10, 20, 40, 80 μM) or DHA/HTF for 48 h. The fluorescent probe DCFH-DA identified a dose-dependent increase in ROS following exposure to DHA. Furthermore, combined exposure of Jurkat cells to 20 μM DHA and 20 μM HTF produced over three-fold more ROS than exposure to 20 μM DHA alone (p < 0.001). The results represented mean ± standard deviation. ***p<0.001, compared with the control group, # p<0.001, compared with DHA.
Fig 3.
DHA and DHA/HTF promote Jurkat cell apoptosis.
Jurkat cells (1 × 106/well) were exposed to DHA (0, 10, 20, 40, 80 μM) or DHA/HTF for 48 h, and measured apoptosis by Annexin V/PI staining and flow cytometry. DHA induced a dose-dependent increase in apoptosis (a). This difference was statistically significant (p < 0.001). DHA/HTF induced a 2.75-fold increase in apoptosis relative to control (p < 0.001). The results represented mean ± standard deviation (b). ***p<0.001, compared with the control group, # p<0.001, compared with DHA.
Fig 4.
DHA and DHA/HTF impaired cell cycle progression in Jurkat cells.
Jurkat cells (1 × 106/well) were exposed to DHA (0, 10, 20, 40, 80 μM) or DHA/HTF for 48 h, Cell cycle analysis of DHA- and DHA/HTF-treated cells was performed using flow cytometry. The ratio of cells in S and G2/M phases decreased while the numbers in G0/G1 phase increased in a dose-dependent manner (a, b). The results represented mean ± standard deviation. ***p<0.001, compared with the control group, # p<0.001, compared with DHA.
Fig 5.
Treatment of Jurkat cells with DHA or DHA/HTF inhibited TfR mRNA expression.
Jurkat cells (1 × 106/well) were exposed to DHA (0, 10, 20, 40, 80 μM) or DHA/HTF for 48 h, Treatment of Jurkat cells with DHA induced a dose-dependent decrease in TfR expression (a). Compared with DHA treatment, DHA/HTF treatment decreased TfR expression by 43.21% (p < 0.001). Agarose gel electrophoresis showed that PCR products were of the expected size (b). GAPDH: 258 bp; TfR: 83 bp. The results represented mean ± standard deviation. ***p<0.001, compared with the control group, # p<0.001, compared with DHA.
Fig 6.
Treatment of Jurkat cells with DHA or DHA/HTF inhibited VEGF mRNA expression.
Jurkat cells (1 × 106/well) were exposed to DHA (0, 10, 20, 40, 80 μM) or DHA/HTF for 48 h, treatment of Jurkat cells with DHA induced a dose-dependent decrease in VEGF expression (a). Compared with DHA treatment, DHA/HTF treatment decreased VEGF expression by 47.77% (p < 0.001). Agarose gel electrophoresis showed that PCR products were of the expected size (b). GAPDH: 258 bp; VEGF: 91 bp. The results represented mean ± standard deviation. ***p<0.001, compared with the control group, # p<0.001, compared with DHA.
Fig 7.
Treatment of Jurkat cells with DHA or DHA/HTF inhibited telomerase activity.
Jurkat cells (1 × 106/well) were exposed to DHA (0, 10, 20, 40, 80 μM) or DHA/HTF for 48 h, treatment of Jurkat cells with DHA induced a dose-dependent decrease hTERT expression (a). Compared with DHA treatment, DHA/HTF treatment decreased hTERT expression by 31.13% (p < 0.001). Agarose gel electrophoresis showed that PCR products were of the expected size (b). GAPDH: 258 bp; hTERT: 134 bp. The results represented mean ± standard deviation. ***p<0.001, compared with the control group, # p<0.001, compared with DHA.