FIGURE 3.

CD13 Overexpression promotes ROS scavenging genes transcription. (A) RNA transcriptome chip analysis on the mRNA levels of vector control and CD13 overexpression PLC/PRF/5 cells. ANPEP indicates CD13. (B) The mRNA levels of SOD1, GPX1, GPX2 and GPX3 analyzed in vector control and PLC/PRF/5/CD13 cells by qRT-RCR. (C–F) qRT-PCR on mRNA levels of ROS scavenging genes in PLC/PRF/5 cells (C) treated with H₂O₂ (12.5, 25 μM) for 24 h (D,E) treated with 5FU (200 μM) combined with NAC (500 μM) or ubenimex (200 μM) (F) treated with combination of 5FU and ubenimex. Data are presented as mean ± SEM (n = 3). *p ≤ 0.05, ***p ≤ 0.001. (G) CD13 and NRF1 expression was determined by using Immunoblot. A representative immunoblot from three independent experiments giving similar results is shown and β-actin was used as a protein loading control. (H) PLC/PRF/5 cells were infected with lentivirus. After puromycin screening, PLC/PRF/5 cells exhibiting stable CD13 overexpression or knockdown were obtained. Immunoblot was performed to detect CD13 and NRF1 expression. A representative immunoblot from three independent experiments giving similar results is shown and β-actin was used as a protein loading control. (I,J) qRT-PCR on mRNA expression of NRF1 and ROS scavenging genes in PLC/PRF/5 cells transfected with NRF1-siRNA and treated with 200 μM 5FU for 24 h. Data are presented as mean ± SEM (n = 3). *p ≤ 0.05, ***p ≤ 0.001.